Mixture toxicity of six pharmaceuticals towards Aliivibrio fischeri, Daphnia magna, and Lemna minor.

As the knowledge on the joint effects of pharmaceuticals towards different non-target organisms is still limited, the aim of our study was to evaluate the toxicity of mixtures of pharmaceuticals, as well as their baseline toxicity towards three selected organisms, namely the bioluminescent bacteria Aliivibrio fischeri, the crustacean Daphnia magna, and the duckweed Lemna minor. Different mixtures composed of three up to five pharmaceuticals having the same or different mechanisms of action in terms of their therapeutic activity (non-steroidal anti-inflammatory drugs, opioid analgesic, antibacterial and anti-epileptic drugs) were investigated. The observed EC50s were compared with those predicted using the concentration addition (CA) and independent action (IA) models. In general, the EC50 values for mixtures predicted with the CA model were lower than those obtained with the IA model, although, in some cases, test predictions of these two models were almost identical. Most of the experimentally determined EC50 values for the specific mixtures were slightly higher than those predicted with the CA model; hence, a less than additive effect was noted. Based on the obtained results, it might be concluded that the CA model assumes the worst-case scenario and gives overall closer predictions; therefore, it should be recommended also for modeling the mixture toxicity of pharmaceuticals with different modes of action.

Multifunctional compounds in the extract from mature seeds of Vicia faba var. minor: Phytochemical profiling, antioxidant activity and cellular safety in human selected blood cells in in vitro trials.

BACKGROUND: The field bean (Vicia faba) is a valuable fodder plant of the Fabaceae family, grown as a main crop for its seed yield. Its phytochemical profile is characterized by the presence of a range of compounds with various biological activities. PURPOSE: The present study investigates the phytochemical profile of the extract from mature seeds of Vicia faba var. minor and examines its impact on preventing oxidative damage to various lipids, protein and DNA molecules in vitro. METHODS: Human plasma was treated with H2O2/Fe (an OH. donor) to induce oxidative damage to lipids and proteins, and the plant extract was then added. As oxidative stress may influence the biological activity of plasma, e.g. coagulation, and influence cardiovascular disease, the study also examined the effect of the plant extract on coagulation and monoamine oxidase activity (MAO, EC 1.4.3.4). RESULTS: The tested extract exerted a protective effect on plasma lipids and proteins treated with H2O2/Fe. However, while it appears to effectively protect the DNA in peripheral blood mononuclear cells from oxidative damage, it did not induce changes in the coagulation process, and significantly reduced MAO activity when applied at 1, 5, and 10 µg/mL. It is possible that the observed antioxidant potential may be due to the complex chemical composition of the extract: the phytochemical profile demonstrated a range of phenolic compounds, including catechins. CONCLUSION: Our findings indicate that extract from mature seeds of V. faba var. minor may be a promising source of antioxidants in multiple applications, including diseases associated with oxidative stress; however, more studies based on in vitro and in vivo models are needed to determine its biological properties.

Evaluation of Antimicrobial, Antioxidant, and Cytotoxic Activity of Phenolic Preparations of Diverse Composition, Obtained from Elaeagnus rhamnoides (L.) A. Nelson Leaf and Twig Extracts.

Although the major components of various organs of sea buckthorn have been identified (particularly phenolic compounds), biological properties of many of these phytochemicals still remain poorly characterized. In this study, we focused on the chemical composition and biological activity of preparations that were obtained from sea buckthorn twigs and leaves. The objective was to investigate cytotoxicity of these preparations against human fibroblast line HFF-1, using MTT reduction assay, their anti- or pro-oxidant activities against the effects of a biological oxidant -H2O2/Fe-on human plasma lipids and proteins in vitro (using TBARS and carbonyl groups as the markers of oxidative stress). Antimicrobial activity of the tested preparations against Gram-positive (Staphylococcus aureus, S. epidermidis, Enterococcus faecalis) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), as well as against fungi (Candida albicans, C. glabrata) by the EUCAST-approved broth microdilution method, followed by growth on solid media, were also assessed. Our analysis showed significant differences in chemical composition and biological properties of the tested preparations (A-F). All tested preparations from sea buckthorn twigs (D-F) and one preparation from sea buckthorn leaves (preparation C) may be a new source of phenolic antioxidants for pharmacological and cosmetic applications.

Ecotoxicity screening evaluation of selected pharmaceuticals and their transformation products towards various organisms.

The intensive development of medical science has led to an increase in the availability and use of pharmaceutical products. However, nowadays, most of scientific attention has been paid to the native forms of pharmaceuticals, while the transformation products (TPs) of these substances, understood herein as metabolites, degradation products, and selected enantiomers, remain largely unexplored in terms of their characterization, presence, fate and effects within the natural environment. Therefore, the main aim of this study was to evaluate the toxicity of seven native compounds belonging to different therapeutic groups (non-steroidal anti-inflammatory drugs, opioid analgesics, beta-blockers, antibacterial and anti-epileptic drugs), along with the toxicity of their 13 most important TPs. For this purpose, an ecotoxicological test battery, consisting of five organisms of different biological organization was used. The obtained data shows that, in general, the toxicity of TPs to the tested organisms was similar or lower compared to their parent compounds. However, for example, significantly higher toxicity of the R form of ibuprofen to algae and duckweed, as well as a higher toxicity of the R form of naproxen to luminescent bacteria, was observed, proving that the risk associated with the presence of drug TPs in the environment should not be neglected.

Response of blood platelets to phenolic fraction and non-polar fraction from the leaves and twigs of Elaeagnus rhamnoides (L.) A. Nelson in vitro.

Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) is a shrub with lanceolate leaves and orange fruits. In traditional Chinese medicine, sea buckthorn organs, especially fruits have been used to treat different diseases, for example cardiovascular disorders. In vitro studies indicate that the phenolic-rich fraction of sea buckthorn fruits demonstrates anti-platelet activity. The aim of the present study was therefore to determine the influence of phenolic and non-polar fractions isolated from the leaves and twigs of sea buckthorn on various parameters of human blood platelets in vitro. Plant material consisted of four different fractions: (1) the phenolic fraction isolated from the leaves, (2) the phenolic fraction isolated from the twigs, (3) the non-polar fraction isolated from leaves and (4) the non-polar fraction from twigs. The chemical composition of the tested fractions was determined using reversed phase UHPLC-HRMS/MS. The fractions from twigs were found to have stronger anti-platelet properties than those from leaves, and all tested fractions were found to be safe for the blood platelets. The tested fractions from the sea buckthorn, especially the non-polar fraction from the twigs, may potentially be a source of compounds with antiplatelet activity.

Feline hip joint anatomy in magnetic resonance images.

The aim of this study was to develop an anatomical model of the feline hip joint for low-field magnetic resonance imaging (LF-MRI) based on high-field magnetic resonance imaging (HF-MRI). The study was performed on six adult clinically healthy European shorthair cats, aged 1-3 years, with body weight of 2.8-4.4 kg. The animals were examined with the use of the Vet-MRI Grande Esaote LF (0.25 T) scanner and high-field Siemens Magnetom TRIO (3 T) MRI scanner. In the LF-MRI, most satisfactory results in T1-weighted images were obtained when TE was 26 ms in all three planes and when TR was 350-950 ms in the transverse plane, 950-1150 ms in the sagittal plane and 520-750 ms in the dorsal plane. In T2-weighted images, TE was 90 ms in the transverse and dorsal plane and 120 ms in the sagittal plane. The results were presented as images acquired with LF-MRI scanners in three planes. The slice thickness was 3 mm for each plane. In LF-MRI, muscles in the hip joint region and round ligament were well visualized. Unlike in LF-MRI, the cross section of the femoral nerve was identified in HF-MRI scans. In examinations of the feline hip joint, the main limitations of LF-MRI were a lack of reliable contrast between articular cartilage and synovial fluid as well as longer scan time. Despite the above, LF-MRI images were characterized by good contrast between bones and the surrounding soft tissues.

Biological properties of Elaeagnus rhamnoides (L.) A. Nelson twig and leaf extracts.

BACKGROUND: Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson, SBT) is a valuable plant because of its medical and therapeutic potential. Different bioactive compounds in SBT berries are of special interest to various researchers. However, not only sea buckthorn berries, but also leaves of this plant (both fresh and dried) contain a lot of nutrients and bioactive compounds, including phenolic compounds. The present study was carried out in order to investigate antioxidant and anticoagulant properties of sea buckthorn twig and leaf extracts (0.5-50 µg/mL) by using various in vitro models. Moreover, the aim of present experiments was to compare the biological activity of SBT leaf extract and SBT twig extract with selected berry extracts (a rich source of phenolic compounds): SBT berry extract (flavonoids being the dominant components), a commercial extract from the berries of Aronia melanocarpa (Aronox®), and a grape seed extract. METHODS: We determined the effect of plant extracts on the oxidative stress using selected markers of this process, i.e. the level of carbonyl groups in proteins. Additionally, we analysed the potential mechanism of modulation of hemostatic properties of human plasma (using selected coagulation times). RESULTS: SBT twig and leaf extracts were observed to exhibit an antioxidant activity against two strong biological oxidants: hydrogen peroxide (H2O2) and H2O2/Fe (the donor of hydroxyl radicals), which induced human plasma lipid peroxidation and protein carbonylation. Both extracts also showed anticoagulant properties. CONCLUSIONS: Our present results have demonstrated that extracts from different parts of SBT, especially berries and twigs, in comparison to well-known berries (aronia and grape), may also be viewed as a good source of active substances - antioxidants for pharmacological or cosmetic applications. Moreover, it is very important from an economic point of view to know that there is a possibility of obtaining phenolic compounds not only from the berries or leaves, but also from twigs, which constitute a production waste.

The Influence of Haemostatic Dressing Prototypes for the Emergency Services on the Histopathological Parameters of Porcine Muscle.

BACKGROUND/AIM: Haemostatic dressings for the uniformed and rescue services are an integral part of life-saving equipment for controlling post-traumatic haemorrhage. The aim of this study was to assess the influence of active constituent substances and materials of haemostatic dressings on muscle tissue and muscle regeneration after traumatic injury. MATERIALS AND METHODS: Three hemostatic dressing prototypes were analysed: OBR/G/S sponge: dressing material sponge made of Na-Ca chitosan/algal composite microfibers and nanofibers; OBR/MBT/S: tactic gauze modified with a polymer mixture of Na-Ca chitosan/algal composite microfibers and nanofibers, impregnated with a moderate amount of procoagulants (22.9 g/m2); and OBR/MS/S: seton gauze modified with a polymer mixture of Na-Ca chitosan/algal composite microfibers and nanofibers, impregnated with a moderate amount of procoagulants (18.0 g/m2), with chitosan (ChitoClearhqg 95) and sodium alginate (Protanal LF10/60 FT) as the coagulants. The experiment was conducted on 20 pigs which were euthanised 24 h, 7 or 14 days after wound dressing. Samples of porcine muscle tissue were subjected to qualitative histopathological analysis. RESULTS: Histopathological analysis of muscle tissues from the experimental pigs revealed that the application of modified seton (OBR/MS/S) produced the most satisfactory results. The observed changes were similar on all dates that samples were collected and in all experimental groups, and minor differences in their extent were observed between groups. Regenerative processes were most advanced, and retrograde changes were least apparent in animals treated with OBR/MS/S. CONCLUSION: Modified seton (OBR/MS/S) induced the least tissue reaction and was most effective in promoting tissue regeneration after injury.

Migration of human mesenchymal stem cells stimulated with pulsed electric field and the dynamics of the cell surface glycosylation.

BACKGROUND: The analysis of the stem cells' glycome dynamics at different stages of differentiation and migration makes possible the exploration of the cell surface glycans as markers of the stem cell functional status, and, in the future, compatibility between transplanted cell and host environment. OBJECTIVES: The objective of our study was to develop novel techniques of investigating cell motility and to assess whether the electric field of the therapeutic spinal cord stimulation system used in vivo contributes to the migration of human mesenchymal stem cells (hMSCs) in vitro. MATERIAL AND METHODS: We have investigated the electrotaxis of bone marrow-derived MSCs using pulsed electric field (PEF) in the range of 16-80 mV/mm and the frequency of 130 Hz and 240 Hz. The PEF-related dynamics of the cell surface glycosylation was evaluated using 6 plant lectins recognizing individual glycans. RESULTS: Pulsed electric field at physiological levels (10 mV/mm; 130 Hz) did not influence cellular motility in vitro, which may correspond to the maintenance of the transplanted cells at the lesion site in vivo. An increase of the PEF intensity and the frequency exceeding physiological levels resulted in an increase in the cellular migration rate in vitro. Pulsed electric field elevated above physiological intensity and frequency (40-80 mV/mm; 240 Hz), but not at physiological levels, resulted in changes of the cell surface glycosylation. CONCLUSIONS: We found the described approach convenient for investigations and for the in vitro modeling of the cellular systems intended for the regenerative cell transplantations in vivo. Probing cell surface glycomes may provide valuable biomarkers to assess the competence of transplanted cells.

Comparative chemical composition, antioxidant and anticoagulant properties of phenolic fraction (a rich in non-acylated and acylated flavonoids and non-polar compounds) and non-polar fraction from Elaeagnus rhamnoides (L.) A. Nelson fruits.

This study focuses on two fractions from sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) fruits: the phenolic fraction (rich in non-acylated and acylated flavonoids and non-polar compounds) and the non-polar fraction. The objective was to investigate both the chemical composition of these fractions, as well as their biological activities in vitro. The tested fractions of sea buckthorn inhibited lipid peroxidation induced by H2O2, however, the non-polar fraction reduced more powerfully the process induced by H2O2/Fe as compared to the phenolic fraction. The tested fractions of sea buckthorn fruits also inhibited carbonylation stimulated by H2O2/Fe. Moreover, the action of the phenolic fraction and non-polar fraction on hemostatic parameters of plasma was also compared to activities of other phenolic fraction, in which flavonoids were the dominant compounds. Our results indicate that sea buckthorn fruits are a rich source of different secondary metabolites, i.e. triterpenes and their derivates, which possess not only antioxidant properties, but may also display anticoagulant attributes.

Disseminated Pancreatic Neuroendocrine Neoplasm (NEN) with an Uncommon Localisation in the Central Nervous System. A Case Report.

BACKGROUND: Neuroendocrine neoplasms (NEN) are rare neoplasms that originate from neuroendocrine cells and are characterized by the potential of hormonal activity. Approximately 70% of these tumours are located in the gastrointestinal system (GI), followed by the bronchi, endocrine glands-like C cells of the thyroid (medullary carcinoma), the parasympathetic and sympathetic system (paragangliomas, pheochromocytoma) and other very rare locations. The prevalence of cerebral metastases in neuroendocrine tumours is estimated by various authors to be approximately 1.5-5%. When the primary tumour is located in the pancreas, it is associated with a risk of cerebral metastases lower than 2%. CASE REPORT: We describe a patient with a disseminated pancreatic NEN that presented with an isolated lesion in the brain. We gathered the important data via medical history,, observation, analysis of medical records, imaging and others diagnostic tests. Despite the fairly rare prevalence of cerebral metastases in NENs, a neurological work-up should be performed. This should include neuroimaging of the brain, preferably with MR, together with the somatostatin receptor scintigraphy (SRS), in each clinically suspicious case. A histopathological examination of the CNS tumour can confirm a dedifferentiation of NEN in the direction of a neuroendocrine carcinoma (NEC - neuroednocrine carcinoma) with a poor prognosis. CONCLUSIONS: Cerebral metastases are diagnosed in 1.5-5% of patients with a neuroendocrine neoplasm. In each case suggestive of a dissemination into the central nervous system, MRI of the brain should be performed.