Salivary VEGF and post-extraction wound healing in type 2 diabetic immediate denture wearers.

OBJECTIVE: Oral wound healing in healthy could be promoted by VEGF in saliva, and immediate denture wearing, but data in type 2 diabetes are lacking. Aims were to investigate the timeline of extraction wound healing in diabetic participants wearing immediate dentures and its correlation to salivary VEGF, as well as to examine the impact of the palatal plate on tissue VEGF during palatal wound healing in rat diabetic model. MATERIAL AND METHODS: Healthy (42) and type 2 diabetic (36) denture wearers, candidates for teeth extractions were included. Extraction wound healing was followed via measurements of socket closure, gingival hyperaemia, pain and presence of necrosis on 3rd, 7th, 14th and on 21st-day post-extraction. Salivary VEGF was measured before and on the 3rd and 21st day after the extraction. In streptozotocin-induced diabetic (30) as well as non-diabetic rats (30), tissue VEGF was measured in palatal wounds healing under or without a palatal plate. RESULTS: Type 2 diabetic prosthetic patients exhibit delayed socket closure, with pronounced hyperaemia, pain and necrosis. Salivary VEGF is increased in diabetes and positively correlates to socket closure while negatively with pain on 21st day after the extraction. Palatal incision induced VEGF increase in non-diabetic and diabetic, but less pronounced in diabetic rats. Wound healing under the palatal plate exhibit higher tissue VEGF. CONCLUSION: Type 2 diabetes-induced increase in salivary VEGF may mitigate diabetes-induced detrimental effects on extraction wound healing. Lack of adequate tissue VEGF response to injury may underly dysregulation of diabetic oral wound healing.

Efficacy of the hatching event in assessing the embryo toxicity of the nano-sized TiO2 particles in zebrafish: a comparison between two different classes of hatching-derived variables.

The aim of the present study was to evaluate the nano-TiO2 toxicity to zebrafish embryos through evaluating the success in hatching in relationship with hours post-exposure instead of considering just the total hatching rate. Zebrafish embryos 4h post-fertilization were exposed to nTiO2 (0, 0.01, 10, and 1000 µg mL(-1)) for 130 h. The hatching rate (HR) was calculated for each concentration (treatment). The HR magnitude was significantly (p<0.001) correlated (using simple regression) to hours post-exposure time interval (hpe; 34, 58, 82, 106, and 130), noted as HR.hpe. The HR descriptive statistics (HRds) and the parameters of the regression models (i.e., constant, x, F, and r(2)) were recruited to define 15 HRds- and 4 h.hpe-derived variables, respectively. The efficacy of the variables was evaluated. Exposure to nTiO2 led to a significant: premature hatching and general decrease in time required for normal hatching; and change in HR and hpe interrelations in a dose-dependent manner. The major change in hatchability between the treatment and control occurred at 58 hpe (62 hpf), when the treatment with nTiO2 induced significant premature hatching compared to only 6% of the hatched embryos in the control at the same time point. EC10 and EC50 values that cause premature hatching at 58 hpe for nTiO2 are 0.073 µg mL(-1) and 107.2 µg mL(-1) respectively. In general(,) this study shows multivariate differences among exposure concentrations of nTiO2 recruiting hatching-derived endpoints.

Antimicrobial, antioxidative, and insect repellent effects of Artemisia absinthium essential oil.

In this paper, the chemical composition and biological activity of the essential oil of Artemisia absinthium was studied. The aim of this study was to investigate the potential of ethnopharmacological uses of this plant species in the treatment of gastrointestinal diseases and wounds, and as an insect repellent. The aerial part of the plant was hydrodistilled, and the chemical composition of the essential oil was analyzed by gas chromatography and gas chromatography/mass spectrometry. Forty-seven compounds, corresponding to 94.65 % of the total oil, were identified, with the main constituents being sabinene (24.49 %), sabinyl acetate (13.64 %), and α-phellandrene (10.29 %). The oil yield was 0.23 % (v/w). The antimicrobial activity of the oil was investigated against ten bacterial isolates (from patients wounds and stools) and seven American Type Culture Collection strains using a microwell dilution assay. The minimal inhibitory/bactericidal concentration of the oil ranged from < 0.08 to 2.43 mg/mL and from 0.08 to 38.80 mg/mL, respectively. The antioxidant activity of the essential oil was evaluated using 2,2-diphenyl-1-picrylhydrazil and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical-scavenging methods and assessed as significant. Skin irritation potential and acute toxicity of the oil were also investigated. Results of the skin irritant reaction showed that none of the 30 volunteers developed a positive skin irritant reaction to undiluted A. absinthium essential oil. Acute oral exposure to the essential oil did not cause mortality in the treated mice, but it did cause neurological, muscle, and gastrointestinal problems. A subchronic toxicity test on Drosophila melanogaster showed that the essential oil of A. absinthium is toxic for developing insect larvae. Starting with the concentration of 0.38 % of essential oil in medium, significant mortality of larvae exposed to the oil was noted when compared to the control. Probit analysis revealed that the LC50 value of A. absinthium essential oil for D. melanogaster larvae after 15 days of exposure was 6.31 % (49 mg/mL). The essential oil also affected the development of D. melanogaster larvae and significantly delayed achievement of the pupa stadium.

Changes in the wing shape and size in fruit flies exposed to micro and nanoplastics.

Geometric morphometrics analysis (GMA) is a well-known technique to identify minute changes in Drosophila wings. This study aimed to determine potential changes in Drosophila wings shape and size after exposure to polystyrene nanoplastics (NPs) (50 nm) and microplastics (MPs) (1 µm). Flies were exposed from eggs to pupal eclosion and analyzed using GMA. Results revealed a difference in shape and size between male and female wings, as expected, due to sexual dimorphism. Therefore, wings were analyzed by sex. Wings of MPs and NPs treated females were elongated compared to controls and had a constriction of the wing joint. Additionally, MPs treated female flies had the most dissimilar shape compared to controls. In male flies, NPs flies had smaller wings compared to MPs and control flies. Compared to control, NPs wings of males were shrunken at the joint and in the entire proximal region of the wing. However, male MPs wings had a narrower anal region and were slightly elongated. These results reveal that wing shape and size can change in a different way based on the sex of the flies and size of plastic particles that larvae interacted with. All the changes in the wings occurred only within the normally allowed wing variation and treatment with NPs/MPs did not cause development of the aberrant phenotypes. Results can pave the way for further understanding of how MPs and NPs can alter phenotypes of flies.

The heart of plastic: utilizing the Drosophila model to investigate the effects of micro/nanoplastics on heart function.

Microplastics (MPs) and nanoplastics (NPs) have increasingly been found in the environment. Until recently, most MPs/NPs toxicological research has been done in aquatic systems resulting in a gap in knowledge regarding terrestrial systems. Plastics have been shown to enter the circulatory system of humans, and can accumulate within organs, little is known about the effect this has on health. Heart disease is the leading cause of death globally, so it's critical to understand the possible impacts MPs/NPs have on the heart. The Drosophila model has been growing in popularity within the toxicology field, it allows for affordable and rapid research on the impacts of a variety of toxins, including plastics. Some research has examined toxicological effects of plastics on the fly, evaluating the effects on mortality, fecundity, development, and locomotion. However, no one has studied the effects on the Drosophila heart. We utilize the Drosophila model to identify the potential effects of oral exposure to polystyrene MPs (1 µm in diameter) and NPs (0.05 µm in diameter) particles on heart function. Flies were exposed to 1.4 × 1011 particles/d/kg of larvae for MPs and 1.2 × 1018 particles/d/kg of larvae for NPs from egg to pupal eclosion. Heart function was then analyzed utilizing semi-intact dissections and Semi-automatic Optic Heartbeat Analysis software (SOHA). Following exposure to MPs and NPs we see sexually dimorphic changes to heart size and function. This study highlights the importance of additional Drosophila MPs/NPs research to identify the molecular mechanisms behind these changes.

Beneath the surface: Decoding the impact of Chironomus riparius bioturbation on microplastic dispersion in sedimentary matrix.

A detailed understanding of microplastics (MPs) behaviour in freshwater ecosystems is crucial for a proper ecological assessment. This includes the identification of significant transport pathways and net accumulation zones, considering their inherent, and already proven influence on aquatic ecosystems. Bioavailability of toxic agents is significantly influenced by macroinvertebrates' behaviour, such as bioturbation and burrowing, and their prior exposure history. This study investigates the effect of bioturbation activity of Chironomus riparius Meigen, 1804 on the vertical transfer of polyethylene MPs ex-situ. The experimental setup exposes larvae to a scenario of 10× the environmentally relevant high concentration of MPs (80 g m-2). Bioturbation activity was estimated using sediment profile imaging with luminophore tracers. This study demonstrated that spherical MPs are vertically transferred in the sediment due to the bioturbation activity of C. riparius larvae and that their presence influences the intensity of the bioturbation activity over time. The present findings provide a noteworthy contribution to the understanding of the relationship between ecosystem engineers and the dispersion and accumulation of MPs within freshwater ecosystems.

Histopathology of chironomids exposed to fly ash and microplastics as a new biomarker of ecotoxicological assessment.

In the last few decades, industrial pollution has gained extensive attention in terms of its effect on the aquatic environment. This imposes the need to develop sensitive biomarkers for early detection of pollutant toxicity in ecotoxicological assessment. The advantages of histopathological biomarkers are many, including quick reaction to the presence of contaminants, and the small number of individuals needed for efficient analysis. The present study analyzed the negative effect of lignite coal fly ash (LCFA) and microplastic particles (MPs) on Chironomus riparius, a suggested model organism by the Organization for Economic Cooperation and Development (OECD). This study aimed to perform histological analyses of larval tissues and target potential changes in treated groups that could serve as promising histopathological biomarkers of the contaminant's negative effects. Following that, other known sensitive sub-organismal biomarkers were analyzed and paired with the histopathological ones. Histological analysis of larvae showed a significantly decreased length of microvilli in midgut regions II and III in both treatments. Treatments with MPs affected oxidative stress parameters: thiobarbituric acid reactive substances (TBARS), advanced oxidation protein products (AOPP), superoxide dismutase (SOD), and hemoglobin levels, while LCFA significantly affected all tested sub-organismal biomarkers (DNA damage, levels of AOPP, SOD, and hemoglobin), except catalase (CAT) and TBARS. When observing histological slides, a significant shortage of brush border length in the posterior parts of the midgut was detected in all treatments. In the case of LCFA, the appearance of intensive vacuolization of digestive cells with inclusions resembling apoptotic bodies, in mentioned regions was also detected. This study demonstrated high sensitivity of brush border length to the MPs and LCFA exposure, complementary to other tested sub-organismal biomarkers. Revealing the great potential of this histopathological biomarker in ecotoxicological studies contributes to the international standard ecotoxicology assessment of emerging pollutants.

Toxic effects of a cyanobacterial strain on Chironomus riparius larvae in a multistress environment.

Cyanobacteria and their toxic metabolites present a global threat to water habitats, but their impact on aquatic organisms in a multistress environment has been poorly investigated. Here we present the results of a survey on the effects of the toxic cyanobacterial strain Trichormus variabilis (heterotypic synonym Anabaena variabilis), and its toxic metabolite, cyanotoxin microcystin-LR, on Chironomus riparius larvae in a multistress environment. An environmentally relevant concentration of microcystin-LR (0.01 mg/L) caused an increase in larvae mortality in an acute toxicity test, which became greater in the presence of environmental stressors (NO3-, NH4+, PO43- and Cd2+), pointing to an additive effect of these agents. Chronic exposure of C. riparius larvae to the microcystin-LR producing strain of T. variabilis in a multistress environment led to a reduction in the larval mass and hemoglobin concentration, and it induced DNA damage in larval somatic cells. The results revealed the additive effect of microcystin-LR in combination with all three tested stressors (NO3-, NH4+, PO43-), and the deleterious effect of chronic exposure of C. riparius larvae to the microcystin-LR producing T. variabilis in a multistress environment. However, the present study further emphasizes the importance of investigating interactions between stressors and cyanotoxins, and their effect on aquatic organisms.

In Situ Effects of a Microplastic Mixture on the Community Structure of Benthic Macroinvertebrates in a Freshwater Pond.

Benthic communities contain some of the most threatened organisms in aquatic habitats due to different anthropogenic pressures. The high abundance of microplastics in sediments will continue to increase in the future, further increasing the probability of interactions between macroinvertebrates and microplastics. In the present study, a benthic community in a relatively pristine shallow pond was exposed either to an environmentally relevant high concentration of a microplastic mixture of 80 g m-2 in the sediment, or a control sediment, without the addition of microplastics. The mixture of microplastics contained irregularly shaped polyethylene, polyvinyl chloride, and polyamide in a ratio of 50:25:25%, respectively. The in situ experiment lasted for 100 d. The total number of taxa that colonized the microcosms was 22 (17 in the control and 18 in the microplastic treatment), and the colonization was not affected by the treatment. The most dominant group within the macroinvertebrate community was the dipteran family Chironomidae, in both the control and the microplastic treatment. No significant differences in the abundance and biomass at a community level were recorded between the groups by permutational multivariate analysis of variance (F = 0.993, p = 0.456 and F = 0.344, p = 0.797, respectively). The mixture of microplastics did not influence the abundance or biomass of the functional feeding groups (F = 1.810, p = 0.137 and F = 0.377, p = 0.736, respectively). The species richness, species abundance, species biomass, Shannon's diversity index, and Simpson's index of diversity showed no statistically significant differences between the control and treatment groups. Czekanowski's quantitative similarity index indicated that 84% of the community remained unaffected after microplastic exposure. Environ Toxicol Chem 2022;41:888-895. © 2021 SETAC.

Effects of a microplastic mixture differ across trophic levels and taxa in a freshwater food web: In situ mesocosm experiment.

The ubiquitous presence of microplastics (MP) in aquatic ecosystems can affect organisms and communities in multiple ways. While MP research on aquatic organisms has primarily focused on marine ecosystems and laboratory experiments, the community-level effects of MP in freshwaters, especially in lakes, are poorly understood. To examine the impact of MP on freshwater lake ecosystems, we conducted the first in situ community-level mesocosm experiment testing the effects of MP on a model food web with zooplankton as main herbivores, odonate larvae as predators, and chironomid larvae as detritivores for seven weeks. The mesocosms were exposed to a mixture of the most abundant MP polymers found in freshwaters, added at two different concentrations in a single pulse to the water surface, water column and sediment. Water column MP concentrations declined sharply during the first two weeks of the experiment. Contrary to expectations, MP ingestion by zooplankton was low and limited mainly to large-bodied Daphnia, causing a decrease in biomass. Biomass of the other zooplankton taxa did not decrease. Presence of MP in the faecal pellets of odonate larvae that fed on zooplankton was indicative of a trophic transfer of MP. The results demonstrated that MP ingestion varies predictably with MP size, as well as body size and feeding preference of the organism, which can be used to predict the rates of transfer and further effects of MP on freshwater food webs. For chironomids, MP had only a low, short-term impact on emergence patterns while their wing morphology was significantly changed. Overall, the impact of MP exposure on the experimental food web and cross-ecosystem biomass transfer was lower than expected, but the experiment provided the first in situ observation of MP transfer to terrestrial ecosystems by emerging chironomids.

Gene expression of zebrafish embryos exposed to titanium dioxide nanoparticles and hydroxylated fullerenes.

Increased release of engineered nanoparticles to the environment suggests a rising need for the monitoring and evaluation of potential toxicity. Zebrafish frequently have been used as a model species in human and aquatic toxicology studies. In this study, zebrafish embryos were microinjected in the otic vesicle with a sublethal dose of engineered nanoparticles (titanium dioxide/TiO(2) and hydroxylated fullerenes/C(60)(OH)(24)). A gene microarray analysis was performed on injected and control embryos to determine the potential for nanoparticles to change the expression of genes involved in cross talk of the nervous and immune systems. The exposure to TiO(2) and hydroxylated fullerenes caused shifts in gene regulation response patterns that were similar for downregulated genes but different for upregulated genes. Significant effects on gene regulation were observed on genes involved in circadian rhythm, kinase activity, vesicular transport and immune response. This is the first report of circadian rhythm gene deregulation by nanoparticles in aquatic animals, indicating the potential for broad physiological and behavioral effects controlled by the circadian system.

Effects of nanosized titanium dioxide on innate immune system of fathead minnow (Pimephales promelas Rafinesque, 1820).

Effects of nanosized (<100 nm) titanium dioxide (TiO(2)) particles on fish neutrophils and immune gene expression was investigated using the fathead minnow (Pimpehales promelas). Expanded use of TiO(2) in the cosmetic industry has increased the potential exposure risk to aquatic ecosystems and human health. Effects of nano-TiO(2) on neutrophil function of the fathead minnow was investigated using oxidative burst, neutrophil extracellular traps (NETs) release and degranulation of primary granules. The innate immune gene expression was determined with quantitative PCR (qPCR). Application of 0.1 µg mL(-1) of nano-TiO(2) in vitro stimulated oxidative burst and NET release. Intraperitoneal injection of 10 µg g(-1) of nano-TiO(2) caused a significant decrease in oxidative burst, NETs release and degranulation (21%; 11%; and 30%, decrease, respectively). Fish exposed to nano-TiO(2) for 48 h in vivo had significantly increased expression of interleukin 11, macrophage stimulating factor 1, and neutrophil cytosolic factor 2 (4; 2.5; and 2 fold increase, respectively). Nano-TiO(2) has potential to interfere with the evolutionary conserved innate immune system responses, as evidenced with observed changes in gene expression and neutrophil function. This finding encourages the use of fish models in the studies of nanoparticle immunotoxicity. The lowest significant response concentration studied in vitro is four times greater than the estimated environmental concentration for TiO(2) (0.025 µg mL(-1)) causing concern about potential impact of nano-TiO(2) on aquatic animals and ecosystems.

From nanoplastic to microplastic: A bibliometric analysis on the presence of plastic particles in the environment.

Identifying leading publications, authors, and countries in microplastic and nanoplastic research is beneficial for regulatory decisions, determining standardized research methodology, and solidifying definitions. Here, bibliometric analysis was performed using Web of Science's Core Collection to evaluate publication trends. A total of 3820 publications were downloaded and analyzed with the majority being journal articles. Since 2009, the number of publications has substantially increased. Results revealed that although the USA and China are the topmost publishing countries, two out of three of the top publishing institutions lie outside of these countries. The year with highest total number of citations was 2019 (42,000 citations), followed by 2018 (25,000 citations) and 2017 (13,000 citations). The journal Marine Pollution Bulletin published the highest number of records and included the top cited publications. Top publishing countries and the top cited publications and authors will likely pave the way for standardization in both microplastic and nanoplastic research.

A Multiparametric Approach to Cerium Oxide Nanoparticle Toxicity Assessment in Non-Biting Midges.

Cerium oxide nanoparticles (CeO2 NPs) are included in the Organisation for Economic Co-operation and Development (OECD) priority list of engineered nanomaterials for assessment of their environmental impact. The present study was carried out to assess the CeO2 NP toxicity to the freshwater midge Chironomus riparius larvae at concentrations of 2.5, 25, 250, and 2500 mg of CeO2 NP/kg of sediment. Experiments were designed to assess the prolonged exposure of midges to CeO2 NPs while adhering to OECD test guideline 218. The following parameters were investigated: CeO2 NP uptake by larvae, oxidative stress parameters, in vivo genotoxic effects, and life trait parameters. Inductively coupled plasma-mass spectrometry analysis showed a significant positive correlation between the concentration of CeO2 NPs in the sediment and its uptake by the larvae. No significant mortality was observed in C. riparius, and oxidative stress was not detected. The only significantly induced sublethal effect was genotoxicity, which began to manifest at a lowest-observed-effect concentration of 25 mg kg-1 of sediment and progressively increased at higher concentrations. Our results indicate that exposure to CeO2 NP-contaminated freshwater sediments does not pose a risk to chironomids at environmentally realistic concentrations. However, the significant accumulation of CeO2 NPs by chironomid larvae may pose a risk through trophic transfer to organisms further up the food chain. Environ Toxicol Chem 2019;39:131-140. © 2019 SETAC.

Changes in the wing shape and size in Drosophila melanogaster treated with food grade titanium dioxide nanoparticles (E171) - A multigenerational study.

Drosophila is among the most commonly used models for toxicity assessment of different types of nanoparticles. This study aims to examine the effects of a constant exposure to the low concentration of human food grade titanium dioxide nanoparticles (TiO2 E171) on Drosophila melanogaster wing morphology over multiple generations. Subsequently, the Geometric Morphometrics Analysis was employed to examine possible changes in the wing shape and size of the treated flies. The treatment resulted in the diminishment but not a disruption in the sexual dimorphism in wings. Consequently, the female flies were clearly separated from the male flies by the differences in wing morphology as in the control group. A splitting by generations was overly similar within the control and the treatment, but it was slightly more pronounced in the treatment. However, the observed generational differences seemed mostly random between generations, irrespective of the treatment. Specifically, the treated groups displayed slightly higher splitting by generations in females than in males. Regardless of the generation, the results show a clear splitting by the differences in the wing shape between the treated flies and the flies from control. The mean value of centroid size, which refers to the wing size, of both female and male wings was smaller in the treatment when compared to the control. The overall effect of TiO2 was to induce significant difference in Drosophila wing morphology but it did not alter the general wing morphology pattern. Therefore, the change in the wings occurred only within the normally allowed wing variation.

Can phytoplankton blooming be harmful to benthic organisms? The toxic influence of Anabaena sp. and Chlorella sp. on Chironomus riparius larvae.

Cyanobacteria and microalgae are abundant biota groups in eutrophic freshwater ecosystems, serving as a food source for many aquatic organisms, including the larvae of non-biting midges (Chironomidae). Many species of cyanobacteria are toxin producers, which can act as stressors to other organisms. The present study aimed to analyze and compare the effects of dietary exposure to the common toxic cyanobacteria Anabaena sp. and non-toxic microalgae Chlorella sp. in Chironomus riparius larvae. Microcystin was detected and quantified in the methanolic extract of Anabaena sp. using the HPLC-DAD technique, and it was identified as microcystin-LR. Both Anabaena sp. and Chlorella sp. were suitable food sources to enable the survival of C. riparius larvae in laboratory conditions, causing negligible mortality and significant differences in the larval mass (ANOVA and Post hoc LSD test; p < 0.05) and hemoglobin concentration (Student's t-test; p < 0.05). Oxidative stress parameters such as advanced oxidation protein products (AOPP), thiobarbituric acid reactive substances (TBARS), catalase (CAT) and superoxide dismutase (SOD) activity, and DNA damage, were also investigated. One-way ANOVA, followed by the Post hoc LSD test, showed a significant increase in AOPP and CAT for the group of larvae fed with Chlorella sp. The same test showed moderate DNA damage in both groups of larvae, with greater damage in the group fed with Anabaena sp. Thus, Chlorella sp. and microcystin-LR producing Anabaena sp. are food sources that did not result in any drastic acute effect on the population level of C. riparius larvae. However, sub-individual-level endpoints revealed significant effects of the treatments, since they caused oxidative stress and DNA damage that may pose a danger to successive generations of test organisms.

Exposure to a microplastic mixture is altering the life traits and is causing deformities in the non-biting midge Chironomus riparius Meigen (1804).

The effect of microplastics (MP) exposure on the chironomid species Chironomus riparius Meigen, 1804 was investigated using the OECD sediment and water toxicity test. Chironomid larvae were exposed to an environmentally relevant low microplastics concentration (LC), a high microplastics concentration (HC) and a control (C). The LC was 0.007 g m-2 on the water surface + 2 g m-3 in the water column + 8 g m-2 in the sediment, and the HC was 10 X higher than this for each exposure. The size of the majority of the manufactured microplastic pellets varied between 20 and 100 µm. The MP mixture consisted of: polyethylene-terephtalate (PET), polystyrene (PS), polyvinyl-chloride (PVC) and polyamide (PA) in a ratio of 45%: 15%: 20%: 20%, respectively, for the sediment exposure; 100% polyethylene for the water column exposure; and 50% polyethylene: 50% polypropylene for the water surface exposure. Different endpoints were monitored, including morphological changes in the mandibles and mentums of 4th instar larvae, morphological changes in the wings, mortality, emergence ratio, and developmental time. A geometric morphometric analysis showed a tendency toward widening of the wings, elongation of the mentums and changing the shape of the mandibles in specimens exposed to both concentrations of microplastics. The development time of C. riparius was significantly prolonged by the MP treatment: 13.8 ± 0.5; 14.4 ± 0.6; and 15.3 ± 0.4 days (mean ± SD) in the C, LC, and HC, respectively. This study indicates that even environmentally relevant concentrations of MP mixture have a negative influence on C. riparius, especially at the larval stage.

Phytochemistry, Toxicology and Therapeutic Value of Petasites hybridus Subsp. Ochroleucus (Common Butterbur) from the Balkans.

Petasites hybridus (Common butterbur) is extensively used in traditional medicine, and is currently gaining interest and popularity as a food supplement and for its medicinal properties. It contains a large number of active compounds of potential therapeutic activity, but also toxic pyrrolizidine alkaloids. Science-based information is needed to support the developing modern use of this plant, and to direct continued safe practice in traditional medicine. The present study focused on the essential oils from leaves and rhizomes of the understudied P. hybridus ssp. ochroleucus from the Balkans, and evaluated its phytochemistry and potential therapeutic activities (antimicrobial, antioxidant, anti-cholinesterase and anti-inflammatory), as well its toxicology potential (acute toxicity in insects and mice). We studied the essential oils, which are not commonly used in traditional practices, but have a potential for safe use since the toxic pyrrolizidine alkaloids, which are non-volatiles, are usually not present in the distilled essential oils. Pyrrolizidine alkaloids were indeed not detected in the essential oils; ingestion of the essential oils did not induce toxicity signs in mice, and topical application did not elicit skin irritation in humans. The essential oils had no antimicrobial properties against 20 pathogenic bacterial strains, but demonstrated good local anti-inflammatory activity in a Carrageenan-induced paw edema test. An insect toxicity test demonstrated that the leaf essential oil is an efficient insect repellent, and the demonstrated anti-cholinesterase activity suggests a potential for the treatment of neurological conditions. Isopetasin, a sesquiterpene found in plants of the genus Petasites, known to have anti-inflammatory effects, was present only in the rhizomes essential oil (3.9%), and sesquiterpene lactones concentrations were high, likely contributing to the antioxidant activity.

In silico prediction of MicroRNA role in regulation of Zebrafish (Danio rerio) responses to nanoparticle exposure.

The release of nanoparticles to the environment can affect health of the exposed organisms. MicroRNAs have been suggested as potential toxicology biomarkers, however the information about use of microRNA in aquatic organisms exposed to nanoparticles (NP) is limited. In silico analysis from publicly available gene expression data was performed. Data selection for the analysis was based on reported biological and pathological outcomes of NP induced toxicity in zebrafish. After identifying relevant genes, we constructed six miRNA-mRNA regulatory networks involved in nanoparticle induced toxicological responses in zebrafish. Based on our prediction and selection criteria we selected six miRNAs that overlapped in constructed networks with remarkable prediction score, and were validated by previous mammalian and zebrafish microRNA profiling studies: dre-miR-124, -144, -148, -155, -19a, -223. The results of this in silico analysis indicate that several highly conserved miRNAs likely have a regulatory role of organismal responses to nanoparticles, and can possibly be used as biomarkers of nanotoxicity in studies using zebrafish as model organism One health approaches.

The effects of a human food additive, titanium dioxide nanoparticles E171, on Drosophila melanogaster - a 20 generation dietary exposure experiment.

In this study, fruit flies (Drosophila melanogaster) were exposed to an estimated daily human E171 consumption concentration for 20 generations. Exposure to E171 resulted in: a change in normal developmental and reproductive dynamics, reduced fecundity after repetitive breeding, increased genotoxicity, the appearance of aberrant phenotypes and morphologic changes to the adult fat body. Marks of adaptive evolution and directional selection were also exhibited. The larval stages were at a higher risk of sustaining damage from E171 as they had a slower elimination rate of TiO2 compared to the adults. This is particularly worrisome, since among the human population, children tend to consume higher daily concentrations of E171 than do adults. The genotoxic effect of E171 was statistically higher in each subsequent generation compared to the previous one. Aberrant phenotypes were likely caused by developmental defects induced by E171, and were not mutations, since the phenotypic features were not transferred to any progeny even after 5 generations of consecutive crossbreeding. Therefore, exposure to E171 during the early developmental period carries a higher risk of toxicity. The fact that the daily human consumption concentration of E171 interferes with and influences fruit fly physiological, ontogenetic, genotoxic, and adaptive processes certainly raises safety concerns.