Beneficial Effects of Laurel (Laurus nobilis L.) and Myrtle (Myrtus communis L.) Extract on Rat Health.

Polyphenols of Laurel and Myrtle exhibit structural diversity, which affects bioavailability, metabolism, and bioactivity. The gut microbiota plays a key role in modulating the production, bioavailability and, thus the biological activities of phenolic metabolites, particularly after the intake of food containing high-molecular-weight polyphenols. The aim of this study was to investigate whether the polyphenolic components of Laurel and Myrtle aqueous extract have beneficial effects on rat health. The growth of lactic acid bacteria (LAB), ß-glucuronidase, ß-glucosidase, ß-galactosidase activity, pH value, body weight change and food efficacy ratio after intragastric treatment of rats with Laurel and Myrtle extract at doses of 50 and 100 mg/kg for two weeks were investigated. The endogenous populations of colonic probiotic bacteria (Lactobacilli and Bifidobacteria) were counted on selective media. According to the obtained data, Laurel extract in the applied dose of 50 and 100 and Myrtle extract (100 mg/kg) positively affects the rats health by increasing the number of colonies of Lactobacilli and Bifidobacteria compared to the control group, causes changes in glycolytic enzymatic activity and minor change in antioxidative tissue activity. In addition, high doses of Laurel increase food efficiency ratio, while Myrtle has the same effect at a lower dose.

Blackthorn Flower Extract Impact on Glycaemic Homeostasis in Normoglycaemic and Alloxan-Induced Hyperglycaemic C57BL/6 Mice.

RESEARCH BACKGROUND: The use of plants and their extracts in treatments of chronic diseases is widely known in traditional medicine. The aim of this study is to determine the effects of 10-day consumption of blackthorn (Prunus spinosa L.) flower extract on blood glucose, glycaemic load, serum α-amlyase activity and insulin concentration in normoglycaemic and hyperglycaemic (alloxan-induced) mice model. EXPERIMENTAL APPROACH: Normoglycaemic and hyperglycaemic (treated with alloxan, 150 mg per kg body mass) C57BL/6 mice were administered daily, during 10 days, blackthorn flower extract by gavage. The sugar mass concentration within the extract was determined by HPLC analysis. In mice, blood and serum blood glucose concentrations, and oral glucose tolerance test were determined by blood glucometer. Serum insulin concentration was determined by ELISA assay and α-amylase activity by colourimetric assay. RESULTS AND CONCLUSIONS: The blackthorn flower extract increased glucose concentrations in normoglycaemic mice by 30% after the 1st and 5th day and by 17% after the 10th day of consumption. It is a consequence of released sugars because sugar analysis revealed 59.8 mg/L monosaccharides, mainly fructose (55.7 mg/L) and glucose (24.3 mg/L) in the extract. On the contrary, the extract consumption reduced serum blood glucose in hyperglycaemic mice by 29% after 10 days of treatment. Oral glucose tolerance test also confirmed that in the hyperglycaemic group treated with blackthorn flower extract glucose homeostasis was improved and showed decrease in blood glucose. Serum insulin concentration increased by 49% and serum α-amylase activity by 46% after 10 days of treatment with blackthorn flower extract in hyperglycaemic group. Thus, it can be concluded that blackthorn flower extract improved glucose tolerance, enhanced insulin secretion and lowered serum α-amylase activity. NOVELTY AND SCIENTIFIC CONTRIBUTION: The obtained results show for the first time the potential of blackthorn (Prunus spinosa L.) flower extract in hyperglycaemia management.

Enhanced activity of superoxide dismutase is a common response to dietary and genetically induced increased cholesterol levels.

Objectives: Hypercholesterolaemia has been implicated in the pathogenesis of neurodegenerative diseases. In this work, we tested whether cholesterol-mediated neurodegeneration induced either by cholesterol-rich diet or genetic mutation may share a common mechanism involving increased oxidative stress and mitochondria oxidant status. Additionally, we analysed whether upon cholesterol-rich diet, different brain regions (prefrontal cortex, cortex, hippocampus, and cerebellum) show distinct vulnerability to an oxidative stress response.Methods: Oxidative stress parameters were measured both in vivo (in the liver and in different brain regions) in cholesterol-fed mice and in vitro in genetically induced cholesterol accumulation in NPC1-null cells.Results: Increased superoxide dismutase (SOD) activity was a common feature of cholesterol-mediated antioxidant response in both models. Moreover, upon high-cholesterol diet, all four brain regions analysed responded via somewhat different capacity of antioxidant defence, hippocampus showing the highest basal activity of SOD. Increased activity of SOD upon cholesterol accumulation in vitro involves mitochondrial SOD2. We found that SOD/SOD2 activities are modulated by cholesterol levels.Discussion: Hypercholesterolaemia could potentiate brain dysfunction and neurodegenerative processes via oxidative stress, and activity of mitochondrial SOD2 may play a key role in this process. Our findings suggest that preventing/reducing mitochondrial oxidative stress may represent a common approach against neurodegenerative diseases.

Polyphenols from Wine Lees as a Novel Functional Bioactive Compound in the Protection Against Oxidative Stress and Hyperlipidaemia.

The study examines the potential of wine industry by-product, the lees, as a rich mixture of natural polyphenols, and its physiological potential to reduce postprandial metabolic and oxidative stress caused by a cholesterol-rich diet in in vivo model. Chemical analysis of wine lees showed that their total solid content was 94.2%. Wine lees contained total phenols, total nonflavonoids and total flavonoids expressed in mg of gallic acid equivalents per 100 g of dry mass: 2316.6±37.9, 1332.5±51.1 and 984.1±28.2, respectively. The content of total anthocyanins expressed in mg of cyanidin-3-glucoside equivalents per 100 g of dry mass was 383.1±21.6. Antioxidant capacity of wine lees determined by the DPPH and FRAP methods and expressed in mM of Trolox equivalents per 100 g was 259.8±1.8 and 45.7±1.05, respectively. The experiment lasted 60 days using C57BL/6 mice divided in four groups: group 1 was fed normal diet and used as control, group 2 was fed normal diet with added wine lees, group 3 was fed high-cholesterol diet (HCD), i.e. normal diet with the addition of sunflower oil, and group 4 was fed HCD with wine lees. HCD increased serum total cholesterol (TC) by 2.3-fold, triacylglycerol (TAG) by 1.5-fold, low-density lipoprotein (LDL) by 3.5-fold and liver malondialdehyde (MDA) by 50%, and reduced liver superoxide dismutase (SOD) by 50%, catalase (CAT) by 30% and glutathione (GSH) by 17.5% compared to control. Conversely, treatment with HCD and wine lees reduced TC and LDL up to 1.4 times more than with HCD only, with depletion of lipid peroxidation (MDA) and restoration of SOD and CAT activities in liver, approximating values of the control. HDL levels were unaffected in any group. Serum transaminase activity showed no hepatotoxic properties in the treatment with lees alone. In the proposed model, wine lees as a rich polyphenol source could be a basis for functional food products without alcohol.

Levels of selected oxidative stress markers in the vitreous and serum of diabetic retinopathy patients.

PURPOSE: In diabetes, an impaired antioxidant defense system contributes to the development of diabetic retinopathy. The main objective of this paper was to find correlations of oxidative stress parameters within and between the vitreous and serum in patients with type 2 diabetes who had developed proliferative diabetic retinopathy. METHODS: The study included and compared two groups of patients who underwent vitrectomy: 37 patients with type 2 diabetes and proliferative retinopathy (PDR), and 50 patients with non-diabetic eye disorders (NDED). Vascular endothelial growth factor (VEGF), advanced oxidized protein product (AOPP), and oxidative stress markers (direct lipid hydroperoxidation (LPO), malondialdehyde (MDA), total superoxide dismutase (SOD), and glutathione (GSH)) were measured in the vitreous and serum of both groups and correlated with one another, between humoral compartments and with gender, age, and serum glucose levels. RESULTS: In the vitreous of PDR patients, VEGF, LPO, and MDA (p<0.05) were increased and SOD values were slightly lowered (p<0.05) than in NDED patients. Vitreous AOPP and GSH showed no differences between the groups. In the serum, AOPP, MDA, and SOD were increased (p<0.05) and VEGF was slightly increased (p<0.05) in the PDR group compared to NDED. With regard to gender, similar changes were recorded for both groups, except for the lower serum MDA in males than females in the NDED group. Advanced age showed no significant effect on changes of measured parameters in the vitreous. In the serum, VEGF was positively correlated (p<0.05) and MDA and SOD negatively correlated (p<0.05) with increasing age. Among measured parameters within and between the vitreous and serum, several correlative links occurred in the PDR group that were not present in the NDED group. The most prominent correlation changes were between serum LPO and vitreal LPO, serum SOD and vitreal LPO, serum LPO and serum SOD, and vitreal VEGF and serum SOD. CONCLUSIONS: Among the selected oxidative stress markers, SOD and LPO were highly correlative in both the vitreous and serum in PDR compared to patients without metabolic disorders. Their correlations suggested that monitoring their mutual alterations might be informative during PDR development and should be considered in further research.

Role of flavonoids on oxidative stress and mineral contents in the retinoic acid-induced bone loss model of rat.

PURPOSE: Reactive oxygen species play a role in a number of degenerative conditions including osteoporosis. Flavonoids as phyto-oestrogens exert physiological effects against oxidative stress diseases. We developed a retinoic acid-induced bone loss model of rats to assess whether flavonoids and alendronate as positive control have role against oxidative stress and mineral contents in osteoporosis in vivo. METHODS: Three-month-old female rats of the Y59 strain were given quercetin, chrysin, naringenin (100 mg kg(-1)) or alendronate (40 mg kg(-1), a positive control) immediately before retinoic acid treatment (80 mg kg(-1)) once daily for 14 days by a single intragastric (i.g.) application. In the second part of the study, we assessed the effect of those flavonoids on the skeletal system of healthy rats using single i.g. application on the respective flavonoids during 14 days. Twenty-four hours after the treatment, we analysed bone mineral density and the total content of bone calcium and phosphorus in the femur, the geometric and physical characteristics of thigh bones and lipid peroxidation and glutathione levels of liver and kidney cells. RESULTS: All flavonoids improved the decrease in bone weight coefficient, the length and the diameter of the bone, the content of bone ash and calcium and phosphorus content induced by retinoic acid. Chrysin and quercetin showed promise as preventive agents. Flavonoids were superior to alendronate according to some criteria. CONCLUSIONS: These results suggest that the dietary flavonoids could reduce retinoic acid-induced oxidative stress and bone loss and that flavonoids may be useful therapeutics for prevention of skeletal diseases.

The possible role of oxidative stress marker glutathione in the assessment of cognitive impairment in multiple sclerosis.

Oxidative stress markers have a distinct role in the process of demyelination in multiple sclerosis. This study investigated the potential correlation of markers of oxidative stress (glutathione [GSH], catalase) with the number of demyelinating lesions and the degree of disability, cognitive deficit, and depression in patients with relapsing-remitting multiple sclerosis (RRMS). Sixty subjects meeting the criteria for RRMS (19 men and 41 women), and 66 healthy controls (24 men, 42 women) were included. In this study, GSH significantly negatively correlated with the degree of cognitive impairment. This is the first study of subjects with RRMS that performed the mentioned research of serum GSH levels on the degree of cognitive damage examined by the Montreal Scale of Cognitive Assessment (MoCA) test. The development of cognitive changes, verified by the MoCA test, was statistically significantly influenced by the positive number of magnetic resonance lesions, degree of depression, expanded disability status scale (EDSS), age, and GSH values. Based on these results, it can be concluded that it is necessary to monitor cognitive status early in RRMS patients, especially in those with a larger number of demyelinating lesions and a higher EDSS level and in older subjects. Also, the serum level of GSH is a potential biomarker of disease progression, which could be used more widely in RRMS.

Combined effects of valproate and naringin on kidney antioxidative markers and serum parameters of kidney function in C57BL6 mice.

Valproate is known to disturb the kidney function, and high doses or prolonged intake may cause serum ion imbalance, kidney tubular acidosis, proteinuria, hyperuricosuria, polyuria, polydipsia, and dehydration. The aim of this in vivo study was to see whether naringin would counter the adverse effects of high-dose valproate in C57Bl/6 mice and to which extent. As expected, valproate (150 mg/kg bw a day for 10 days) caused serum hyperkalaemia, more in male than female mice. Naringin reversed (25 mg/kg bw a day for 10 days) the hyperkalaemia and activated antioxidative defence mechanisms (mainly catalase and glutathione), again more efficiently in females. In males naringin combined with valproate was not as effective and even showed some prooxidative effects.

The Effect of a High-Protein Diet Supplemented with Blackthorn Flower Extract on Polyphenol Bioavailability and Antioxidant Status in the Organs of C57BL/6 Mice.

The health benefits of polyphenols are based on their bioavailability, which is why a significant portion of research focuses on factors that affect their bioavailability. Previous studies suggest that the intake of polyphenols along with macronutrients in food represents one of the key factors influencing the bioavailability of polyphenols and, consequently, their biological activity in the organism. Since polyphenols in the human diet are mainly consumed in food together with macronutrients, this study investigated the in vivo absorption, metabolism, and distribution of polyphenolic compounds from the water extract of blackthorn flower (Prunus spinosa L.) in combination with a protein-enriched diet in the organs (small intestine, liver, kidney) of C57BL/6 mice. The bioaccumulation of polyphenol molecules, biologically available maximum concentrations of individual groups of polyphenol molecules, and their effect on the oxidative/antioxidative status of organs were also examined. The results of this study indicate increased bioabsorption and bioavailability of flavan-3-ols (EC, EGCG) and reduced absorption kinetics of certain polyphenols from the groups of flavonols, flavones, and phenolic acids in the organs of C57BL/6 mice after intragastric administration of the water extract of blackthorn flower (Prunus spinosa L.) in combination with a diet enriched with whey proteins. Furthermore, subchronic intake of polyphenols from the water extract of blackthorn flower (Prunus spinosa L.) in combination with a diet enriched with whey proteins induces the synthesis of total glutathione (tGSH) in the liver and superoxide dismutase (SOD) in the liver and small intestine. The results of this study suggest potential applications in the development of functional foods aimed at achieving the optimal health status of the organism and the possibility of reducing the risk of oxidative stress-related disease.

Intracellular Molecular Targets and Signaling Pathways Involved in Antioxidative and Neuroprotective Effects of Cannabinoids in Neurodegenerative Conditions.

In the last few decades, endocannabinoids, plant-derived cannabinoids and synthetic cannabinoids have received growing interest as treatment options in neurodegenerative conditions. In various experimental settings, they have displayed antioxidative, anti-inflammatory, antiapoptotic, immunomodulatory, and neuroprotective effects. However, due to numerous targets and downstream effectors of their action, the cellular and molecular mechanisms underlying these effects are rather complex and still under discussion. Cannabinoids are able to neutralize free radicals and modulate the production of reactive oxygen species and the activity of antioxidative systems acting on CB1 and CB2 cannabinoid receptors. The activation of CB1 receptors stimulates signaling pathways involved in antioxidative defense and survival (such as the phosphoinositide 3-kinase (PI3K)/Akt, mitogen-activated protein kinase (MAPK), and Nrf2 pathways) and regulates glutamatergic signaling, the activation of N-methyl-D-aspartate (NMDA) receptors, calcium influx, and the induction of Ca2+-regulated signaling cascades, whereas the neuroprotective effects mediated by CB2 receptors are due to the suppression of microglial activation and the release of prooxidative and proinflammatory mediators. This review summarizes the main molecular mechanisms and new advances in understanding the antioxidative and neuroprotective effects of cannabinoids. Because of the plethora of possible pharmacological interventions related to oxidative stress and cannabinoid-mediated neuroprotection, future research should be directed towards a better understanding of the interplay between activated signal transduction pathways and molecular targets with the aim to improve treatment options and efficacy by targeting the endocannabinoid system.

The Effects of Nettle Extract Consumption on Liver PPARs, SIRT1, ACOX1 and Blood Lipid Levels in Male and Female C57Bl6 Mice.

The aim of this study was to evaluate how nettle (Urtica dioica L.) water extract consumption would interact with regulators of peroxysomal lipid oxidation, histone deacetylase, and markers of oxidative stress in the liver and blood lipid levels in male and female C57Bl6 mice. Metabolically unchallenged (healthy) mice (n = 5 per sex) were treated with a nettle extract in a dose of 40 mg of total polyphenols in the extract per kg mice body weight. The nettle extract was applied daily along with normal diet for 15 days. The serum triglycerides, cholesterol, HDL, LDL, and liver PPAR-α, PPAR-γ, PGC-1-α, ACOX1, SIRT1, MDA, SOD, CAT, and GSH were compared between exposed and unexposed (control) animals. In males, the PPAR-α, PGC1-α, and ACOX1 levels together with systemic HDL cholesterol were significantly (p ≤ 0.05) increased while the LDL cholesterol decreased (p ≤ 0.05). In females, no changes in PPAR-α and PGC1-α or serum lipids were noted, but the ACOX1 content in the liver was significantly (p ≤ 0.05) increased. The SIRT1 activity increased (p ≤ 0.05) only in females. In both sexes, the PPAR-γ levels were not significantly (p ≤ 0.05) affected in either sex. The results indicate that nettle plant extract has the potential to modulate selected transcriptional factors and histone deacetylase in vivo, with certain sex differences, which should be studied further in similar models.

Effects of naringin and valproate interaction on liver steatosis and dyslipidaemia parameters in male C57BL6 mice.

Valproate is a common antiepileptic drug whose adverse effects include liver steatosis and dyslipidaemia. The aim of our study was to see how natural flavonoid antioxidant naringin would interact with valproate and attenuate these adverse effects. For this reason we treated male C57BL6 mice with a combination of 150 mg/kg of valproate and 25 mg/kg naringin every day for 10 days and compared their serum triglycerides, cholesterol, LDL, HDL, VLDL, and liver PPAR-alpha, PGC-1 alpha, ACOX1, Nrf2, SOD, CAT, GSH, and histological signs of steatosis. Valproate increased lipid peroxidation parameters and caused pronounced microvesicular steatosis throughout the hepatic lobule in all acinar zones, but naringin co-administration limited steatosis to the lobule periphery. In addition, it nearly restored total serum cholesterol, LDL, and triglycerides and liver ACOX1 and MDA to control levels. and upregulated PPAR-alpha and PGC-1 alpha, otherwise severely downregulated by valproate. It also increased SOD activity. All these findings suggest that naringin modulates key lipid metabolism regulators and should further be investigated in this model, either alone or combined with other lipid regulating drugs or molecules.

Association of chronic inflammation with cardiovascular risk in chronic obstructive pulmonary disease-A cross-sectional study.

Background and Aims: COPD is progressive lung disease with known higher cardiovascular (CV) risk, mainly attributed to smoking of cigarettes as the main etiological factor of disease. The aim of this study was to compare CV risk in patients with COPD to control groups of smokers and non-COPD and to investigate the relation of lung function variables, COPD severity, and smoking with Systemic Coronary Risk Estimation (SCORE) risk calculation, arterial stiffness (AS) values, and biological systemic inflammatory markers. Methods: A total of 208 subjects were included in this study: 61 subjects diagnosed with COPD, 83 smokers without COPD, and 64 nonsmokers without COPD. Medical history and clinical data were recorded, including assessment of pulmonary function and AS, calculation of ankle-brachial index, blood analysis, and CV risk assessment by SCORE risk calculation. Results: Subjects with COPD had significantly higher values of SCORE calculation of risk, central aortic pressure, AS, and markers of systemic inflammation compared to control groups of smokers and nonsmokers without COPD (p < 0.001). Furthermore, statistically significant increase in hs-CRP concentration was found between the COPD group and the control group of non-COPD smokers (p < 0.001), and a statistically significantly higher SCORE calculation was found in the COPD group compared to control groups of smokers and nonsmokers without COPD (p < 0.001). Conclusion: The results of the research support further identification and research of biological markers and simple specific tests such as arteriography that will enable progress in personalized treatment of patients with COPD and better primary and secondary prevention of comorbidities with the aim of improved treatment outcome.

Antioxidant and Anti-Atherogenic Activities of Essential Oils from Myrtus communis L. and Laurus nobilis L. in Rat.

Essential oils (EOs) from aromatic and medicinal plants, such as myrtle (Myrtus communis L.) and Laurel (Laurus nobilis L.), are gaining popularity as a potential ingredient in functional foods and nutraceuticals. This study aims to investigate whether the essential oils (EOs) could be effective in weight control, antioxidative and antilipidemic status of rats by affecting microbiota and its enzymes activity and whether changes in intestinal enzyme activity affect the health of rats. The intragastric application of laurel and myrtle EOs to rats for two weeks affects weight loss, reduces glycolytic activity, lipid parameters (cholesterol, triglycerides, low-density lipoprotein cholesterol (LDL-C) and very low-density lipoprotein cholesterol (VLDL-C)) and atherogenic indicators, leading to cardiovascular protection. Laurel EO can be an excellent candidate for the treatment of drug-induced obesity and related diseases, since it affects lipid metabolism in the liver and inhibits the enzymes responsible for the metabolism of carbohydrates into glucose in the digestive tract, leading to weight loss. In contrast, myrtle EO shows a better antioxidant capacity in most tissues, except kidneys, where it causes a pro-oxidative effect, compared to laurel EO. Myrtle EO increases the permeability and instability of the erythrocyte membrane, resulting in a loss of selectivity for the entry of toxic substances into the cell. On the other hand, myrtle EO leads to intestinal inflammation by reducing the number of probiotic bacteria and increasing Enterobacter.

Distribution of major brain gangliosides in olfactory tract of frogs.

Gangliosides are major cell-surface determinants in the central nervous system (CNS) of vertebrates, found both in neuronal and glial cell membranes. Together with cholesterol and glycosylphosphatidylinositol (GPI) - anchored proteins, gangliosides are involved in organization of plasma membrane microdomains. Based on biochemical studies, frog brain was previously described as having low quantities of gangliosides and their distribution pattern in specific brain regions was unknown. Using highly specific monoclonal antibodies generated against four major brain gangliosides (GM1, GD1a, GD1b and GT1b), we examined the distribution of these molecules in CNS of four different species of frogs (Rana esculenta, Rana temporaria, Bufo bufo and Bufo viridis). We also studied the distribution of myelin- associated glycoprotein (MAG), an inhibitor of axonal regeneration, which is a ligand for gangliosides GD1a and GT1b. Our results show that ganglioside GDla is expressed in neurons of olfactory bulb in all studied animals. In the brain of Rana sp., GD1a is expressed in the entire olfactory pathway, from olfactory bulbs to amygdala, while in Bufo sp. GD1a is restricted to the main olfactory bulb. Furthermore, we found that most of myelinated pathways in frogs express MAG, but do not express GD1a, which could be one of the reasons for better axon regeneration of neural pathways after CNS injury in amphibians in comparison to mammals.

Hyperbilirubinemia as an Indicator of Perforated Acute Appendicitis in Pediatric Population: A Prospective Study.

Background: This prospective cohort study aimed to investigate the association of hyperbilirubinemia with perforated appendicitis in the pediatric population. Patients and Methods: A total of 284 children in whom the diagnosis of acute appendicitis was established were included in this study. The patients were allocated in study groups in regard to operative findings. The first study group included patients who had perforated appendicitis (n = 64; 22.5%) whereas the patients in the second group had simple appendicitis (n = 220; 77.5%). Blood samples for serum bilirubin levels and acute inflammatory markers were taken before the patients underwent surgery. The primary outcome of the study was to investigate whether the level of serum bilirubin should be used to distinguish between simple and perforated appendicitis. Results: The median level of serum bilirubin in children with perforated appendicitis was 27 mcmol/L whereas the patients with simple appendicitis had lower median levels of serum bilirubin (10 µmol/L; p < 0.001). An area under the receiver operating characteristic (ROC) curve for total serum bilirubin was 0.876 (95% confidence interval [CI], 0.820-0.929) in the patients who had a perforated appendicitis. An ROC analysis showed the best sensitivity (92%) and specificity (77.3%) for a cutoff value of 15.5 mcmol/L for total serum bilirubin (p < 0.001). Hyperbilirubinemia at admission was found in 35 patients (54.7%) with complicated appendicitis and in 14 patients (6.4%) with non-perforated appendicitis (p < 0.001). The modeling of collected data by multivariable logistic regression identified serum bilirubin concentration (odss ratio [OR] = 1.12; 95% CI, 1.07-1.18; p < 0.001), serum sodium concentration (OR = 0.64; 95% CI, 0.51-0.81; p < 0.001), body temperature (OR = 2.48; 95% CI, 1.05-0.84; p < 0.001), and duration of symptoms (OR = 1.06; 95% CI, 1.02-1.09; p < 0.001) as risk factors for perforated appendicitis. Conclusion: Elevateds level of total serum bilirubin may be useful as an indicator of perforated appendicitis in children. Levels of bilirubin in serum is an inexpensive, simple, and available laboratory marker and should therefore be recommended in the initial evaluation for acute appendicitis in pediatric patients.

The Risk Assessment of Pesticide Ingestion with Fruit and Vegetables for Consumer's Health.

Pesticides are chemicals used in agriculture to protect crops from pests. In addition to protection during cultivation, they are also used after harvesting to extend the shelf life of products. Postharvest control stands out, especially when it comes to products imported from distant countries, resulting in increased concentration of pesticides and risk to human health consuming such products. In this study, analyses of pesticide residues were performed on 200 samples of fruits and vegetables. Pesticide residues were identified and quantified in 30 out of 200 samples. Study results revealed imazalil to be the most frequently detected pesticide. Risk assessment was performed on the obtained results, and it was carried out separately for adults and for children under 6 years of age. Imazalil showed the highest ARfD percentage for adults (max% ARfD 251%), and these values were especially high on risk assessment for children, where they amounted up to max% ARfD 1087%. The study of imazalil impact was performed on 16 Swiss albino mice divided into two groups and 4 subgroups. Experimental group animals were treated with the corresponding NOAEL dose of imazalil (10 mg/kg) for 28 days. Body weight was measured before each pesticide application on a digital electronic Sartorius scale. Peripheral blood analysis was performed after 28-day animal exposure to pesticides. Animals were anesthetized, blood samples were obtained by cardiac puncture, and red blood cell (RBC) count, hemoglobin (Hb) concentration, and white blood cell (WBC) count were determined by standard hematological methods. The organs for determination of imazalil concentration were extracted immediately upon animal sacrifice and stored in a freezer at -80°C until analysis. Results show difference in gain weight, and an increase in WBC count was recorded in the experimental group as compared with a control group of animals. The highest imazalil levels were recorded in adipose tissue (45.2‰) which proves tendency to accumulate.

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.

Prevention of peritoneal carcinomatosis in mice with combination hyperthermal intraperitoneal chemotherapy and IL-2.

PURPOSE: The purpose of this study was to investigate the effect of local chemoimmunotherapy and hyperthermal intraperitoneal chemotherapy (HIPEC) in a mouse model of induced peritoneal carcinomatosis. MATERIAL AND METHODS: Peritoneal carcinomatosis in mice was produced by intraperitoneal implantation of MCa cells (5 x 10(3)). Interleukin-2 (4.1 x 10(4) IU/mouse) was injected into the abdominal cavity of mice at day 7 and 3 before implantation of tumour cells. Immediately after implantation of MCa cells mice were treated twice with 2 ml of saline that was heated either at 37 degrees C or 43 degrees C and cytostatics (doxorubicin 20 mg kg(-1), cisplatin 10 mg kg(-1), mitomycin 5 mg kg(-1), or 5-FU 150 mg kg(-1)). We followed the survival of animals and side effects appearing with different forms of treatment. RESULTS: Combined treatment with Interleukin-2 (IL-2) and cytostatics (5-FU, CIS or MIT) significantly affected the development of peritoneal carcinomatosis and increased the survival of mice (ILS% - 37 degrees C = 29.88, 199.32, and 108.52, ILS% - 43 degrees C = 62.69, 260.50, and 178.05, respectively). However, intraperitoneal chemotherapy on survival time of mice with DOX + IL-2 was ineffective as compared with DOX alone. CONCLUSION: We would like to stress that treatment with IL-2 prior to tumour diagnosis is not clinically practical, rather, the manuscript attempts to describe an experimental proof of principle. Results suggest the synergistic effect of hyperthermia, chemotherapy and immunotherapy; IL-2 significantly increases antitumor activity of hyperthermic chemotherapy and survival rate of mice with peritoneal carcinomatosis.

Effects of prometryne on apoptosis and necrosis in thymus, lymph node and spleen in mice.

Prometryne is a methylthio-s-triazine herbicide. Significant traces are documented in environment, mainly waters, soil and plants used for nutrition. The aim of this study was to estimate prometryne immunotoxic properties through induction of apoptotic and/or necrotic changes in thymocytes, splenocytes and lymph node cells after repeated subchronical exposure. Three different doses of prometryne (185, 375, 555mgkg(-1)) were applied per os every 48h, over 28 days. Flow cytometry assay (annexinV-FITC and PI) was conducted to record apoptotic and necrotic damage. In the spleen significant changes in the percentage of apoptotic cells were not detected between treated and control groups respectively. In thymus and lymph node, within the lowest dose group (185mgkg(-)1), an increase in percentage of early apoptosis without any significant increase in necrosis was detected. Medium (375mgkg(-1)) as well as high dose triggered increase in late apoptosis in lymph node while in thymus; late apoptosis was increased only in animals exposed to the highest dose (555mgkg(-1)). The highest applied dose, in thymus and lymph node respectively, caused a general decrease in percentage of vital cells in favour of marked increase of percentages of all types of dying cells (apoptotic, late apoptotic/early necrotic and necrotic). Prometryne caused disbalance in major organs of immune system, markedly lymph nodes and thymus, by induction of early apoptotic changes in dose/time specific manner.