Development of a classification model for the antigenotoxic activity of flavonoids.

Genotoxic agents are capable of causing damage to genetic material and the cumulative DNA damage causes mutations, involved in the development of various pathological conditions, including cancer. Antigenotoxic agents possess the potential to counteract these detrimental cellular modifications and may aid in preventing, delaying, or decreasing the severity of these pathological conditions. An important class of natural products for which promising antigenotoxic activities have already been shown, are the flavonoids. In this research, we investigated the quantitative structure-activity relationship (QSAR) of flavonoids and their antigenotoxic activity against benzo[a]pyrene (B[a]P) and its mutagenic metabolite B[a]P-7,8-diol-9,10-epoxide-2. Random Forest classification models were developed, which could be useful as a preliminary in silico evaluation tool, before performing in vitro or in vivo experiments. The descriptors G2S and R8s. were the most significant for predicting the antigenotoxic potential.

Insects as an Alternative Source for the Production of Fats for Cosmetics.

Insects may provide an environmentally friendly way of producing high-quality bio-based materials that can be implemented for cosmetic applications. Insects can be bred on organic waste, in high numbers, and on small surfaces, therefore, making large scale industrial breeding possible. Fats from three insect species: the black soldier fly (BSF) (Hermetia illucens), the locust (Locusta migratoria), and the house cricket (Acheta domesticus) were evaluated for potential use in skin care. Insects were dried and fats were extracted using petroleum ether. The fats were further refined, and the fatty acid composition and the acid value were determined. The fats were used in a hand cream formulation and compared with the currently used mink-and plant-derived oils. Fatty acid analysis indicates that BSF contains > 60% of lauric acid, which makes it less suitable for application in a skin-care product, whereas locust and cricket fats are rich in C16 and C18 fatty acids which makes them more suitable. Phospholipids and free fatty acid levels in the three insect species are relatively high compared with commercial, refined oils, and need to be removed by appropriate refining protocols. Odor and color also need to be removed by physical refinement to improve the applicability.

The nutritional profile, mineral content and heavy metal uptake of yellow mealworm reared with supplementation of agricultural sidestreams.

Insect farming, a potential approach to deal with the increasing global protein demand, is a new activity in the Western world with many unanswered questions regarding product quality and safety. Insects may fulfill an important role in a circular economy by upcycling biowaste into valuable biomass. About half of the total mass of mealworm feeding substrates exists out of wet feed. This can be sourced from biowaste, increasing the sustainability of insect farming. This paper reports on the nutritional profile of yellow mealworm, Tenebrio molitor, reared with supplementation of organic sidestreams. These included unsold vegetables, potato cuttings, fermented chicory roots and horticultural foliage. The evaluation was performed by analyzing proximate compositions, fatty acid profiles, mineral and heavy metal contents. Mealworms fed with potato cuttings doubled their fat content and increased saturated and mono-unsaturated fatty acids. Providing fermented chicory roots increased the mineral content and accumulated heavy metals. Additionally, the uptake of minerals by mealworms was selective as only calcium, iron and manganese concentrations increased. Adding vegetable mix or horticultural foliage to the diet did not significantly change the nutritional profile. In conclusion, sidestreams were successfully recycled into protein-rich biomass and their nutrient content and bio-availability influenced the composition of mealworms.

Effect of heat treatment on microbiological safety of supermarket food waste as substrate for black soldier fly larvae (Hermetia illucens).

Supermarket food waste, constituting 13% of global food waste, can be upcycled as substrate for black soldier fly larvae (BSFL) and converted into larval biomass. Since presence of food pathogens in supermarket food waste is likely, microbiological safety should be ensured when using waste as insect substrate. Heat treatment provides a suitable pre-treatment to reduce microbiological contaminations. This study investigated the effect of different temperature-time combinations on the microbiological safety of supermarket food waste as BSFL substrate. Artificial supermarket food waste without meat and fish (SFW) was inoculated with both Salmonella and Staphylococcus aureus (7.0log cfu/g) and treated at 50 and 60 °C for 10, 20 and 30 min. While 50 °C was insufficient for adequate pathogen reduction, 60 °C only required 10 min to reduce the Enterobacteriaceae and S.aureus counts to < 1.0logcfu/g and for absence of Salmonella in 25 g. Heat-treated SFW could be stored for two days at ambient temperature or refrigerated without pathogen growth. Treatment of supermarket food waste containing meat and fish at 60 °C for 10 min caused similar results as for SFW, but S.aureus persisted (2.4logcfu/g), possibly by protective effects of fat and/or proteins. Finally, BSFL rearing experiments on SFW revealed significantly higher larval mass, bioconversion efficiency and waste reduction than on Gainesville diet, with no notable differences between untreated and heat-treated SFW. Rearing BSFL on supermarket food waste is possible, and unsafe food waste can be heated to obtain safety without eliminating nutrients necessary for rearing.

Pilot-Scale Cultivation of the Snow Alga Chloromonas typhlos in a Photobioreactor.

The most studied and cultivated microalgae have a temperature optimum between 20 and 35°C. This temperature range hampers sustainable microalgae growth in countries with colder periods. To overcome this problem, psychrotolerant microalgae, such as the snow alga Chloromonas typhlos, can be cultivated during these colder periods. However, most of the research work has been carried out in the laboratory. The step between laboratory-scale and large-scale cultivation is difficult, making pilot-scale tests crucial to gather more information. Here, we presented a successful pilot-scale growth test of C. typhlos. Seven batch mode growth periods were compared during two longer growth tests in a photobioreactor of 350 L. We demonstrated the potential of this alga to be cultivated at colder ambient temperatures. The tests were performed during winter and springtime to compare ambient temperature and sunlight influences. The growth and CO2 usage were continuously monitored to calculate the productivity and CO2 fixation efficiency. A maximum dry weight of 1.082 g L-1 was achieved while a maximum growth rate and maximum daily volumetric and areal productivities of 0.105 d-1, 0.110 g L-1 d-1, and 2.746 g m-2 d-1, respectively, were measured. Future tests to optimize the cultivation of C. typhlos and production of astaxanthin, for example, will be crucial to explore the potential of biomass production of C. typhlos on a commercial scale.

Impact of Particle Size on Toxicity, Tissue Distribution and Excretion Kinetics of Subchronic Intratracheal Instilled Silver Nanoparticles in Mice.

The unique physicochemical properties of silver nanoparticles (AgNPs) make them useful in a wide range of sectors, increasing their propensity for human exposure, as well as the need for thorough toxicological assessment. The biodistribution of silver, hematological parameters and GSH/GSSG levels in the lung and liver were studied in mice that were intratracheally instilled with AgNP (5 and 50 nm) and AgNO3 once a week for 5 weeks, followed by a recovery period of up to 28 days (dpi). Data was gathered to build a PBPK model after the entry of AgNPs into the lungs. AgNPs could be absorbed into the blood and might cross the physiological barriers and be distributed extensively in mice. Similar to AgNO3, AgNP5 induced longer-lasting toxicity toward blood cells and increased GSH levels in the lung. The exposure to AgNP50 increased the GSH from 1 dpi onward in the liver and silver was distributed to the organs after exposure, but its concentration decreased over time. In AgNP5 treated mice, silver levels were highest in the spleen, kidney, liver and blood, persisting for at least 28 days, suggesting accumulation. The major route for excretion seemed to be through the urine, despite a high concentration of AgNP5 also being found in feces. The modeled silver concentration was in line with the in vivo data for the heart and liver.

Determining the Effect of Different Reproduction Factors on the Yield and Hatching of Tenebrio Molitor Eggs.

Whereas the production of conventional livestock can rely on years of knowledge and experience, the mealworm production industry is still in its early stages. Although the yellow mealworm Tenebrio molitor has been cultivated as feed for pets and zoo animals for quite some time, the optimization of the reproduction process has not been widely explored. For reproduction, beetles are placed in crates supplemented with a substrate to oviposit their eggs. After a specified time, the beetles are removed, and the eggs are further cultivated to develop into larvae. Factors such as oviposition duration and beetle density influence the number of produced mealworms per beetle, partly due to the cannibalistic behavior of the beetles. However, reproductive success has mostly been assessed by determining the number of offspring several weeks after oviposition. As a result, the number of eggs laid and their hatch rates are unknown. In this research, eggs are separated from the oviposition substrate, i.e., harvested. This approach allows to determine the influence of beetle density, oviposition duration, beetle age and the use of a grid during oviposition on the number of eggs produced and the egg hatching rate and timing thereof. In addition, the influence of the harvesting method on the hatching of T. molitor's eggs was determined.

Insects as Diet and Therapy: Perspectives on Their Use for Combating Diabetes Mellitus in Tanzania.

More than 450 million people worldwide are suffering from diabetes and this number is expected to increase. In developing countries, such as Tanzania, the number of patients suffering from diabetes and associated diseases is increasing as well. Up to 80% of the Tanzanian people rely on traditional medicines for their health care services. The nature of Tanzanian is very rich in different plant and insect species, and this could be exploited through their implementation in preventive and/or curative approaches in the battle against diabetes. The implementation of healthy insects in the diets of people may help in the prevention of obesity, which is a risk factor in the etiology of diabetes, while the identification of small molecules in insects may help in the discovery of potential new drugs that can be used in the treatment of diabetes. In this paper, an overview on the potential implementation of insects against diabetes is presented.

Valorisation Potential of Using Organic Side Streams as Feed for Tenebrio molitor, Acheta domesticus and Locusta migratoria.

Due to increasing welfare and population, the demand for alternative protein sources, obtained with minimal use of natural resources, is rising in today's society. Insects have the potential to be used as an alternative protein source since they are considered to be able to convert low-value biomass into high-value components, resulting in opportunities for valorisation of organic side streams. Moreover, insects are suggested to be a sustainable protein source, referring to the efficient "feed to body" mass conversion potential. The aim of this review was to explore the potential to rear the yellow mealworm (Tenebrio molitor), the house cricket (Acheta domesticus) and the migratory locust (Locusta migratoria) on low or not yet valorised organic side streams within the food supply chain. This was performed by collecting research information focusing on the rearing of the insects in scope on organic biomass. In addition, the nutritional composition of the produced insects as well as their dietary requirements will be reviewed. Finally, the availability of side streams in the EU will be discussed as well as their potential to be used as insects feed.

Screening of Tanzanian medicinal plants against Plasmodium falciparum and human immunodeficiency virus.

Medicinal plants used to treat infectious diseases in Bunda district, Tanzania, were screened for activity against Plasmodium falciparum and Human Immunodeficiency Virus Type 1 (HIV-1, IIIB strain) and Type 2 (HIV-2, ROD strain). Antiplasmodial activity was observed for the 80 % MeOH extract of Ormocarpum kirkii (root; MIC = 31.25 microg/mL), Combretum adenogonium (leaves), Euphorbia tirucalli (root), Harrisonia abyssinica (root), Rhynchosia sublobata (root), Sesbania sesban (root), Tithonia diversifolia (leaves), and Vernonia cinerascens (leaves; MIC value of 62.5 microg/mL). With regard to HIV, 80 % MeOH extracts of Barleria eranthemoides (root), Combretum adenogonium (leaves and stem bark), Elaeodedron schlechteranum (stem bark and root bark), Lannea schweinfurthii (stem bark), Terminalia mollis (stem bark and root bark), Acacia tortilis (stem bark), Ficus cycamorus (stem bark) and Indigofera colutea (shoot), as well as H2O extracts from Barleria eranthemoides (root), Combretum adenogonium (leaves and stem bark), and Terminalia mollis (stem bark and root bark) exhibited IC50 values below 10 microg/mL against HIV-1 (IIIB strain). The highest anti-HIV-1 activity value was obtained for the B. eranthemoides 80 % MeOH root extract (IC50 value 2.1 microg/mL). Only a few extracts were active against HIV-2, such as the 80 % MeOH extract from Lannea schweinfurthii (stem bark) and Elaeodedron schlechteranum (root bark), showing IC50 values < 10 microg/mL.

Structural and solvent effects on the 13C and 15N NMR chemical shifts of indoloquinoline alkaloids: experimental and DFT study.

Indoloquinoline alkaloids represent an important class of antimalarial, antibacterial and antiviral compounds. They have been shown to bind to DNA via intercalation preferentially at GC-rich sequences containing nonalternating CC sites. The stability of complexes formed with biological macromolecules depends on noncovalent binding. In the present study, the ability of indoloquinolines to form intermolecular interactions with solvents was investigated by using NMR spectroscopy and density functional theory (DFT) (B3LYP/6-31G**) calculations. NMR data measured for indoloquinoline bases and the corresponding hydrochlorides are discussed in relation to the structure. DFT calculations of shielding constants in vacuo and in solution allowed the investigation of the influence of the environment on the NMR parameters. Calculations incorporating solvent effects indicated significant changes in the anisotropy of the electron distribution, reflected in the span of the chemical shielding tensor (Omega = sigma11 - sigma33). Solvent effects on the span of the 13C and 15N shielding tensor depended on the type of atom and the data indicated a significant influence of solute-solvent interactions.

Evaluation of existing (Q)SAR models for skin and eye irritation and corrosion to use for REACH registration.

The performance of the (Q)SAR models Derek Nexus, Toxtree and Case Ultra for the prediction of skin and eye irritation/corrosion is investigated. For irritation and corrosion of the skin, 117 compounds and for the eye, 125 compounds were listed. The balance between the groups positive and negative for irritation and corrosion was maintained. The obtained predictions were compared with experimental data and the numbers of true and false positives and negatives were determined. Based on these results several performance parameters of the tested (Q)SAR models were calculated. Despite all the efforts to make good and valid models, the results indicate a poor predictivity of the current models: a lot of compounds were not predicted, were out of the applicability domain or were predicted wrong. Considering our results, it can be concluded that the tested models are not yet sufficiently powerful for implementation. Possibly the training-sets used within the current models are not yet comprehensive enough or the incorporated data are not of enough quality. Although the use of these models as stand-alone evaluation is not recommended, these models can be of value as weight-of-evidence in the context of expert knowledge in an Integrated Approach to Testing and Assessment.

(Q)SAR tools for priority setting: A case study with printed paper and board food contact material substances.

Over the last years, more stringent safety requirements for an increasing number of chemicals across many regulatory fields (e.g. industrial chemicals, pharmaceuticals, food, cosmetics, …) have triggered the need for an efficient screening strategy to prioritize the substances of highest concern. In this context, alternative methods such as in silico (i.e. computational) techniques gain more and more importance. In the current study, a new prioritization strategy for identifying potentially mutagenic substances was developed based on the combination of multiple (quantitative) structure-activity relationship ((Q)SAR) tools. Non-evaluated substances used in printed paper and board food contact materials (FCM) were selected for a case study. By applying our strategy, 106 out of the 1723 substances were assigned 'high priority' as they were predicted mutagenic by 4 different (Q)SAR models. Information provided within the models allowed to identify 53 substances for which Ames mutagenicity prediction already has in vitro Ames test results. For further prioritization, additional support could be obtained by applying local i.e. specific models, as demonstrated here for aromatic azo compounds, typically found in printed paper and board FCM. The strategy developed here can easily be applied to other groups of chemicals facing the same need for priority ranking.

Interactions of cryptolepine and neocryptolepine with unusual DNA structures.

Cryptolepine, the main alkaloid present in the roots of Cryptolepis sanguinolenta, presents a large spectrum of biological properties. It has been reported to behave like a DNA intercalator with a preference for GC-rich sequences. In this study, dialysis competition assay and mass spectrometry experiments were used to determine the affinity of cryptolepine and neocryptolepine for DNA structures among duplexes, triplexes, quadruplexes and single strands. Our data confirm that cryptolepine and neocryptolepine prefer GC over AT-rich duplex sequences, but also recognize triplex and quadruplex structures. These compounds are weak telomerase inhibitors and exhibit a significant preference for triplexes over quadruplexes or duplexes.

Synthesis, cytotoxicity, and antiplasmodial and antitrypanosomal activity of new neocryptolepine derivatives.

On the basis of the original lead neocryptolepine or 5-methyl-5H-indolo[2,3-b]quinoline, an alkaloid from Cryptolepis sanguinolenta, derivatives were prepared using a biradical cyclization methodology. Starting from easily accessible educts, this approach allowed the synthesis of hitherto unknown compounds with a varied substitution pattern. As a result of steric hindrance, preferential formation of the 3-substituted isomers over the 1-substituted isomers was observed when cyclizing N-(3-substituted-phenyl)-N'-[2-(2-trimethylsilylethynyl)phenyl]carbodiimides. All compounds were evaluated for their activity against chloroquine-sensitive as well as chloroquine-resistant Plasmodium falciparum strains, for their activity against Trypanosoma brucei and T. cruzi, and for their cytotoxicity on human MRC-5 cells. Mechanisms of action were investigated by testing heme complexation using ESI-MS, inhibition of beta-hematin formation, DNA interactions (DNA-methyl green assay and linear dichroism), and inhibition of human topoisomerase II. Neocryptolepine derivatives with a higher antiplasmodial activity and a lower cytotoxicity than the original lead have been obtained. This selective antiplasmodial activity was associated with inhibition of beta-hematin formation. 2-Bromoneocryptolepine was the most selective compound with an IC(50) value against chloroquine-resistant P. falciparum of 4.0 microM in the absence of cytotoxicity (IC(50) > 32 microM). Although cryptolepine, a known lead for antimalarials also originally isolated from Cryptolepis sanguinolenta, was more active (IC(50) = 2.0 microM), 2-bromoneocryptolepine showed a low affinity for DNA and no inhibition of human topoisomerase II, in contrast to cryptolepine. Although some neocryptolepine derivatives showed a higher antiplasmodial activity than 2-bromocryptolepine, these compounds also showed a higher affinity for DNA and/or a more pronounced cytotoxicity. Therefore, 2-bromoneocryptolepine is considered as the most promising lead from the present work for new antimalarial agents. In addition, 2-bromo-, 2-nitro-, and 2-methoxy-9-cyanoneocryptolepine exhibited antitrypanosomal activity in the micromolar range in the absence of obvious cytotoxicity.

Management of diabetes in Guinean traditional medicine: an ethnobotanical investigation in the coastal lowlands.

ETHNOPHARMACOLOGICAL RELEVANCE: This survey was carried out in the coastal lowlands of Guinea-Conakry in order to make an inventory of plants used by traditional healers, herbalists and diabetic patients for the management of diabetes mellitus. MATERIALS AND METHODS: Frequent ethnomedical and ethnobotanical investigations were conducted from June 2008 to December 2009 in Conakry, Kindia, Forécariah, Dubréka, Boke, Fria and Boffa. It is a cross-sectional survey and data collection is based on the interactive method. During this period a total of 112 people aged from 39 to 76 years old were interviewed. RESULTS: During this investigation 146 plant species belonging to 55 families were collected. The most cited plants were Anacardium occidentale L. and Ficus spp., while the Fabaceae family was the most represented, followed by the Euphorbiaceae and Rubiaceae. The most frequently plant parts used by the traditional healers and the herbalists were the stem-bark and decoctions the most common preparation mode. CONCLUSIONS: It is clear that a variety of plants is used in the management and treatment of diabetes. Due to the increasing prevalence of type 2 diabetes, there is an urgent need for scientific investigations to rationalise the use of these traditional remedies, which could represent accessible alternative medicines for the Guinean populations.

Quality control of roots of Eleutherococcus senticosus by HPLC.

An HPLC method based on several known methods for the determination of eleutherosides B and E was developed, optimised and validated in terms of linearity, precision (repeatability and intermediate precision on different days and at different concentration levels) and accuracy (recovery). The extraction procedure, the extraction solvent and the extraction yield were evaluated and optimised. A reversed-phase RP-18 column gradient eluted with a two-phase system consisting of phosphoric acid:water (0.5:99.5) and acetonitrile was used to evaluate the samples; detection was at 220 nm. Although eleutherosides B and E are commercially available, they are very costly, and therefore ferulic acid was chosen as external standard. The correction factors for the response of ferulic acid against both eleutherosides were determined and validated. This method, accepted by the European Pharmacopoeia Commission, will be included in the monograph on Eleutherococcus senticosus roots to assay the content of eleutherosides B and E.

Antileishmanial activity, cytotoxicity and QSAR analysis of synthetic dihydrobenzofuran lignans and related benzofurans.

A series of synthetic dihydrobenzofuran lignans and related benzofurans were evaluated for their cytotoxicity in a screening panel consisting of various human tumour cell lines, and for their antiprotozoal activity against L. donovani (axenic amastigotes), chloroquine resistant Plasmodium falciparum (strain K1), Trypanosoma brucei rhodesiense and T. cruzi, and for cytotoxicity on L6 cells. No promising cytotoxicities against human tumour cell lines were observed for newly synthesised compounds, but the dimerisation product of some lipophylic esters of caffeic acid, such as compound 2g, showed a high activity against chloroquine-resistant P. falciparum (strain K1) (IC50 0.43 microg/mL) and L. donovani (axenic amastigotes) (IC50 0.12 microg/mL), which was confirmed in an infected macrophage assay (IC50 0.19 microg/mL). QSAR models for the cytotoxic and antileishmanial activity were generated using Quasar receptor surface modelling.

Phytochemical investigation and antioxidant activity of Duranta repens.

The ethylacetate fraction of the methanol extract of Duranta repens L. (Verbenaceae) showed radicalscavenging activity in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Three compounds were isolated from this fraction, i.e. the phenylethanoid glycoside acteoside, the iridoid lamiide and the saponin pseudo-ginsenoside-RT1. Acteoside showed an IC50 of 3.05 +/- 0.09 microg/mL in the DPPH assay, while lamiide and pseudo-ginsenoside-RT1 were not active.

Isoneocryptolepine, a synthetic indoloquinoline alkaloid, as an antiplasmodial lead compound.

The antiprotozoal activities of three naturally occurring isomeric indoloquinoline alkaloids, i.e., cryptolepine (1), neocryptolepine (2), and isocryptolepine (3), and two dimeric indoloquinoline alkaloids, cryptoquindoline (6) and biscryptolepine (7), originally obtained from the plant Cryptolepis sanguinolenta, were compared with those of a new synthetic indoloquinoline isomer, isoneocryptolepine (4), and a quaternary derivative, N-methyl-isocryptolepinium iodide (5). The latter compounds showed a high antiplasmodial activity against the chloroquine-resistant Plasmodium falciparum strain K1 (IC50 of 0.23 +/- 0.04 and 0.017 +/- 0.004 microM, respectively), while the cytotoxicity (L6 cells) was 4.32 +/- 0.04 and 12.7 +/- 2.0 microM, respectively. Isoneocryptolepine (4) was found to act as an inhibitor of beta-hematin formation and as a DNA-intercalating agent.