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This study aimed to estimate the protective effect of a combination of S-Adenosyl-L-methionine (SAM) as a precursor of glutathione and oseltamivir as a specific inhibitor of virus replication on oxidative damages caused by influenza infection in the liver of infected mice. Albino mice were inoculated with 10×LD50 of influenza virus A/Aichi/2/68(H3N2). Oseltamivir was applied for five days after infection, twice per day, in a dose of 2.5 mg/kg. SAM was applied for ten days in a 100 mg/kg dose, starting five days before inoculation. Markers of oxidative stress, mortality rate, mean survival time, index, and protection coefficient were followed. Influenza infection causes severe oxidative damage to the liver. All combinations of SAM and oseltamivir restored the levels of the biochemical markers to those in healthy animals and improved the virological parameters. A combination of SAM 100 mg/kg and oseltamivir 2.5 mg/kg, which is 1/4 of the optimal therapeutic mice dose, exhibits protection index and affects most effectively all the tested parameters. This study provides an easy-to-apply approach with a good therapeutic potential for co-medicating influenza infection with a specific antiviral agent and an antioxidant precursor.

A series of seventeen cinnamic acid hybrids (4ai–ci) were obtained through an amidation of aminoadamantanes (amantadine, rimantadine, and memantine) with mixed anhydride generated from different substituted cinnamic acid and ethyl chloroformate. 1H NMR, 13C NMR, IR, and HRMS were used for the confirmation of the structures of the synthesized hybrids. Moreover, the antioxidant profiles of amides were estimated as per five different in vitro methods: 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid cation radical (ABTS⁺), ferric reducing antioxidant power (FRAP), cupric reducing antioxidant capacity (CUPRAC) assay, and inhibition of Fe(III)/asc induced lipid peroxidation (LP) in brain homogenate. For comparison, caffeic acid (CaffA), known as a potent naturally occurring antioxidant, was used as a reference compound in our study. The results revealed that the most prominent antioxidant activity was demonstrated by compound 4b2, with excellent CUPRAC, FRAP, scavenging ABTS+˙ potential, and inhibition of Fe/asc–induced LP, followed by 4c6 > 4a6 > CaffA > 4c5 and 4a5 > 4a7. Overall, the results suggest that the hybrids (4b2, 4c6, and 4a6) consisting of a caffeoyl moiety and lipophilic adamantane core endow the molecules with the higher antioxidant activity than their parent compound (caffeic acid), especially against LP. Thus, these promising antioxidants could have beneficial effects in various pathological conditions, where oxidative stress is implicated.

Background: Inhibitors of acetylcholinesterase (AChE) are used to treat many disorders, among which are neurodegenerative upsets, like Alzheimer’s disease (AD). One of the limited licensed AChE inhibitors (AChEIs) used as drugs is the natural compound galantamine (Gal). Objective: As Gal is a toxic compound, here we expose data about its four derivatives in hybrid peptide-norgalantamine molecules, which have shown 100 times lower toxicity. Methods: Four newly synthesized galantamine derivatives have been involved in docking analysis made by Molegro Virtual Docker. Biological assessments were performed on ICR male mice. The change in short and long-term memory performance was evaluated by passive avoidance test. AChE activity and levels of main oxidative stress parameters: lipid peroxidation, total glutathione (GSH), enzyme activities of catalase (CAT), superoxide dismutase, and glutathione peroxidase were measured in brain homogenates. Results: Our experimental data revealed that the new hybrid molecules did not impair memory performance in healthy mice. Two of the compounds demonstrated better than Gal AChE inhibitory activity in the brain. None of them changed the level of lipid peroxidation products, one of the compounds increased GSH levels, and all of them increased CAT enzyme activity. Conclusion: The new galantamine-peptide hybrids demonstrated a potential for inhibition of AChE and antioxidant activity and deserve further attention.

Context: The snail hemolymph is a multi-component mixture comprising various substances with pharmacological activity. Aims: To evaluate the antioxidant activity of hemocyanin and fractions isolated of hemolymph from garden snail Helix lucorum. Methods: The antioxidant activity was tested in chemical systems generating superoxide radicals (O2•–) and hydroxyl radicals (•OH) and was presented as percent inhibition. The fractions’ composition was analyzed by MALDI-TOF/TOF mass spectrometry. Results: At concentration of 90 µg/mL the hemolymph fraction with compounds with molecular weight (MW)

Nowadays, an increased interest in natural compounds with preventive or therapeutic potential for various diseases has been observed. Given the involvement of oxidative stress in the pathogenesis of gastric ulcer (GU) and the wide range of bioactive compounds isolated from snails, this study aimed to investigate the protective effect of Cornu aspersum (Müller, 1774) mucus on ethanol-induced GUs. Male albino mice were divided into Control, Ethanol, Mucus + Ethanol and Mucus + Omeprazole treated groups. The GUs were induced by administration of 96% ethanol (10 mL/kg, per os). One hour before ulcer induction, the mice of Mucus + Ethanol group were pretreated with mucus (20 mg/kg, per os), and the mice of Mucus + Omeprazole group were pretreated with omeprazole (20 mg/kg, per os). Ethanol administration caused grave lesions of gastric mucosa and a significant decrease of glutathione (GSH) and superoxide dismutase (SOD), catalase, and glutathione reductase (GR) activities. In the animals with mucus or omeprazole pre-administration compared to the Ethanol group, the following were observed: only a small number of hemorrhagic fields, significantly reduced GU index with calculated 73% protection by mucus and 78% protection by omeprazole, and significant recovery of mucosal GSH and SOD and GR activities. In addition, the mucus inhibited Helicobacter pylori growth. Thus, the protective effect of C. aspersum mucus on both gastric mucosa and gastric antioxidant potential in ethanol-induced GU model suggests that it may serve as a good tool for prevention of this disease.

Alzheimer’s disease (AD) is the most common type of dementia in elderly. Since oxidative stress is an important feature of AD, it could be assumed that conjugation of the available medicines with substances having antioxidant properties may produce beneficial effect. The aim of this study was to investigate the antioxidant capacity of 7 analogs of memantine (MEM) with Gly, Ala, β-Ala, Val, Phe, Phe (4-F), and Gly-Gly. The antioxidant capacity of the MEM conjugates was tested in chemical systems generating superoxide anion (●O2–) and hydroxyl (●OH) radicals, and by applying the DPPH test. The results showed a negligible effect of the substances on DPPH● reduction. Phe-MEM, (4-F)-Phe-MEM and Gly-Gly-MEM demonstrated ●O2– scavenging potential. The Phe-containing derivatives exerted protective effect on ●OH-induced deoxyribose degradation and had abilities for chelating iron ions. In conclusion, the investigated Phe-containing MEM derivatives manifested a good antioxidant activity, and a promising effect in AD treatment could be expected.

Oxidative stress underlies the pathogenesis of many diseases, which determines the interest in natural substances with antioxidant properties.

The aim of the present study was to perform a pilot assessment and analysis of the oxidative stress (OS) level in four commercially important fish species (round goby, red mullet, sprat and horse mackerel) from different localities of the Bulgarian Black Sea coast. The fish were sampled during trawl selectivity experiments. The OS level in the fish was assessed by measuring lipid peroxidation (LPO), glutathione concentration (GSH), activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST), as well as acetylcholine esterase (AChE) in gills and liver. Round goby and red mullet caught in the Nessebar Bay showed clear signs of OS with the highest levels of LPO and GST activities, accompanied by the lowest AChE activities in both liver and gills. On the contrary, round goby caught near Maslen Nos (a region with good ecological conditions) were least affected by OS with low LPO and high GSH concentrations and SOD activity. There were no significant differences in the OS bioindicators of horse mackerel from the different localities. Sprat caught in Nessebar Bay, compared to those caught from the other localities, showed presence of OS indicated by lower GSH levels and relatively higher CAT, GPx and GST activities, accompanied by low AChE activity in gills. It can be concluded that round goby and red mullet were more vulnerable to OS induced by marine environmental factors than the horse mackerel and sprat. However, their antioxidant defense system allows them to tolerate and adapt to the environment of their habitats. Further studies are needed for the assessment of OS in important fish species in the Bulgarian part of the Black Sea.

The wastewater after rose oil distillation is usually discharged into the drainage systems and it represents a serious environmental problem. While being rich in polyphenols, which have beneficial biological activity and application in the pharmaceutical industry, limited research has been carried out about the biological activity of the specific wastewaters per se. Wastewaters after distillation of the four Bulgarian oil-bearing roses Rosa damascena Mill., R. alba L., R. centifolia L., and R. gallica L. exerted significant antioxidant activity and good antiherpes simplex virus type-1 (HSV-1) activity while maintaining a good toxicological safety profile (low cytotoxic effect) towards normal cell lines. More precisely, the non-tumorigenic cells were a human (HEK-293 embryonic kidney cells) and a mouse cell line (CCL-1 fibroblasts, which are recommended as a standard for cytotoxicity evaluation in Annex C of ISO 10993-5). The concentrations that achieved antioxidant and radical scavenging effects (0.04–0.92% v/v) were much lower than most of the maximum tolerated concentrations for the tissue culture cells (0.2–3.4% v/v). The wastewaters had a weak antiproliferative effect against Staphylococcus aureus. None of the wastewaters had activity against Gram-negative bacteria or a bactericidal or antifungal effect. We can conclude that these four species, which are the most preferred species worldwide for producing high-quality rose oil, have the potential to be developed as promising antioxidant and antiherpesvirus nutraceuticals.

This trial studied the influence of environmental conditions (cold, thermoneutral and hot period) on oxidative status of different genotypes free-range laying hens and the effect of oxidative stress (OS) on their egg productive performance. In the experiment, carried with a two-year replication, were used six dual-purpose chicken genotypes with different origins. For determination of the oxidative status were measured the levels of lipid peroxidation (LPO), total glutathione (tGSH) and the activity of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX). The highest egg production for all chicken genotypes was reported during the thermoneutral period. Biochemical parameters LPO and CAT were relatively more sensitive to environmental impact while the remaining (tGSH, SOD, GPX) exhibited a higher degree of stability. The lowest values of LPO for all genotypes were proved during the thermoneutral period. The cold period was most unfavorable – LPO values are elevated for all genotypes. The SOD and GPX values were higher during the cold and the hot periods compared to the thermoneutral. The results of analysis of variance showed that the factor “temperature period” compared to “year” and “genotype” had the most significant influence on all biochemical parameters (P

Small-scale photobioreactors (PBRs) in the inoculum stage were designed with internal (red or green) and external white LED light as an initial step of a larger-scale installation aimed at fulfilling the integral biorefinery concept for maximum utilization of microalgal biomass in a multifunctional laboratory. The specific growth rate of Scenedesmus obliquus (Turpin) Kützing biomass for given cultural conditions was analyzed by using MAPLE software. For the determination of total polyphenols, flavonoids, chlorophyll “a” and “b”, carotenoids and lipids, UHPLC-HRMS, ISO-20776/1, ISO-10993-5 and CUPRAC tests were carried out. Under red light growing, a higher content of polyphenols was found, while the green light favoured the flavonoid accumulation in the biomass. Chlorophylls, carotenoids and lipids were in the same order of magnitude in both samples. The dichloromethane extracts obtained from the biomass of each PBR synergistically potentiated at low concentrations (0.01–0.05 mg/mL) the antibacterial activity of penicillin, fluoroquinolones or oregano essential oil against the selected food-borne pathogens (Staphylococcus aureus, Escherichia coli and Salmonella typhimurium) without showing any in vitro cytotoxicity. Both extracts exhibited good cupric ion-reducing antioxidant capacity at concentrations above 0.042–0.08 mg/mL. The UHPLC-HRMS analysis revealed that both extracts contained long chain fatty acids and carotenoids thus explaining their antibacterial and antioxidant potential. The applied engineering approach showed a great potential to modify microalgae metabolism for the synthesis of target compounds by S. obliquus with capacity for the development of health-promoting nutraceuticals for poultry farming.

Alzheimer’s disease (AD), a progressive neurodegenerative disorder characterized by memory loss and cognitive functions decline, is a leading cause for dementia and currently ranked as the sixth foremost cause of death. As of present, treatment of AD is symptomatic without convincing therapeutic benefits and new, effective, therapeutic agents are pursued. Due to massive loss of cholinergic neurons and decreased acetylcholine levels, cholinesterase inhibitors like galantamine, remain the backbone of pharmacological treatment of the disease. In the present study, using behavioral and biochemical methods, four newly synthesized galantamine derivatives, Gal 34, Gal 43, Gal 44, and Gal 46, were evaluated for a beneficial effect in a scopolamine model of dementia in mice. They were designed to have all the advantages of galantamine and additionally to inhibit β-secretase and exert favorable effects on plasma lipids. Behavioral tests included step-through inhibitory avoidance, T-maze, and the hole-board test, whereas biochemical evaluations involved assessment of acetylcholinesterase activity, brain monoamines levels, lipid peroxidation, catalase, glutathione peroxidase, and superoxide dismutase activities along with measurement of total glutathione. Results show that Gal 43, Gal 44, and, in particular, Gal 46 are especially effective in improving both short- and long-term memory and in the case of Gal 46 having a significant effect on exploratory activity as well. Although Gal 34 did not show behavioral effects as convincing as those of the other three galantamine derivatives, it demonstrated persuasive antioxidant and restorative capacities, making all four galantamine derivatives promising AD treatment agents and prompting further research, especially that in many of our studies they performed better than galantamine.

This study assessed phytochemical profile and antioxidant capacity of hydrosols, obtained by water-steam distillation of Rosa alba L. and Rosa damascena Mill. plants, grown in experimental ...

Geraniol is a common component of essential oils of various plant species and is widely used in cosmetics, perfumery and household products. In the present study, we investigated the cytotoxic, genotoxic and antioxidant activities of different concentrations of the monoterpenoid geraniol and its anti-cytotoxic and anti-genotoxic potential against the genotoxin N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) in higher plant and human lymphocyte test-systems frequently used in genotoxicity studies. The cytogenetic analysis did not show any inhibition of cell viability for geraniol at concentrations 10–100 μg/ml in barley cells. Only low cytotoxicity was observed in human lymphocytes depending on the concentration. No high clastogenicity for human lymphocytes and low enhancement of the frequencies both of chromosome aberrations and micronuclei were detected in barley. Geraniol showed a good radical-scavenging capacity similar to that of butylated hydroxytoluene (BHT), ascorbic acid and α- tocopherol. Geraniol applied at low nontoxic concentrations possessed a clearly expressed genoprotective potential and could inhibit the cytotoxic/genotoxic effects of MNNG reducing both the frequency of chromosome aberrations and micronuclei induced by the genotoxin. The anti-cytotoxic/anti-genotoxic effects were independent of the experimental schemes of treatment and the test-systems. The obtained results will be useful for further investigations concerning the pharmaceutical and medical application of the monoterpenoid geraniol.

A. Alexandrova12, a_alexandrova_bas@yahoo.com, ORCID: 0000-0002-7007-3665, L. Petrov1, dr.lubomir.petrov @gmail.com, ORCID: 0000-0003-1209-959X, R. Makaveev1, racho74@abv.bg, ORCID: 0000-0002-7670-9222, E. Tsvetanova2, elina_nesta@abv.bg, ORCID: 0000-0002-5916-5200, A. Georgieva2, almirageorgieva@gmail.com, ORCID: 0000-0003-1323-2348, S. Kolimechkov1, dr.stefan.kolimechkov@gmail.com, ORCID: 0000-0003-0112-2387 1National Sports Academy, Sofia, Bulgaria, 2Bulgarian Academy of Sciences, Sofia, Bulgaria Александрова Альбена, PhD, доцент кафедры физиологии и биохимии, Национальная спортивная академия; лаборатория свободнорадикальных процессов, Институт нейробиологии, Болгарская академия наук. Болгария, г. София, 1113, ул. Академика Бончева, 23. E-mail: a_alexandrova_bas@yahoo.com, ORCID: 0000-0002-7007-3665. Петров Любомир, PhD, доцент кафедры физиологии и биохимии, Национальная спортивная академия. Болгария, г. София, 1700, ул. Академик Стефан Младенов, 21. E-mail: dr.lubomir.petrov @gmail.com, ORCID: 0000-0003-1209-959X. Макавеев Рашо, PhD, доцент кафедры реслинга и дзюдо, тренерский факультет, Национальная спортивная академия. Болгария, г. София, 1700, ул. Академик Стефан Младенов, 21. E-mail: racho74@abv.bg, ORCID: 0000-0002-7670-9222. Цветанова Элина, PhD, лаборант лаборатории свободнорадикальных процессов, Институт нейробиологии, Болгарская академия наук. Болгария, г. София, 1113, ул. Академика Бончева, 23. E-mail: elina_nesta@abv.bg, ORCID: 0000-0002-5916-5200. Георгиева Альмира, лаборант лаборатории свободнорадикальных процессов, Институт нейробиологии, Болгарская академия наук. Болгария, г. София, 1113, ул. Академика Бончева, 23. E-mail: almirageorgieva@gmail.com, ORCID: 0000-0003-1323-2348. Колимечков Стефан, PhD, научный сотрудник кафедры физиологии и биохимии, Национальная спортивная академия. Болгария, г. София, 1700, ул. Академик Стефан Младенов, 21. E-mail: dr.stefan.kolimechkov@gmail.com, ORCID: 0000-0003-0112-2387. Aim. The aim of this study was to determine the changes in the erythrocyte oxidative status of the wrestlers after performing the maximal aerobic test, by registering in erythrocytes the levels of lipid peroxidation (LPO), total glutathione (tGSH) and activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx). Materials and methods. A group of 12 healthy wrestlers conducted a treadmill maximal aerobic test, and venous blood samples were obtained before and immediately after the exercise. Erythrocytes were separated from plasma and used for spectrophotometric determination of LPO, tGSH and enzyme activities. Plasma was used for determination of hemoglobin concentration (Hb) as an index of hemolysis. Results. The performance of the maximal aerobic test resulted in a significant increase of Hb in blood plasma, a decrease of LPO, and no changes of the tGSH level in erythrocytes. In regards to antioxidant enzymes, our results showed an increase in the activity of GPx, while the CAT and SOD activity remain unchanged. Conclusions. It can be concluded that in active athletes, predominate erythrocytes that are more resistant to oxidative stress, because of the accelerated hemolysis induced by physical exercise, lead to the elimination of the old and oxidative modified cells. Цель. Данное исследование проводилось с целью выявления изменений в оксидатив- ном статусе эритроцитов у борцов после выполнения максимального аэробного теста. В рамках исследования проводилась регистрация уровня пероксидации липидов, общего глутатиона, активности каталазы, супероксиддисмутазы и глутатионпероксидазы. Материалы и методы. 12 здоровых борцов приняли участие в максимальном аэробном тесте на беговой дорожке. Перед тестированием и сразу после выполнения упражнения у участников были взяты образцы венозной крови. Из плазмы крови были выделены эритроциты, в которых методом спектрофотометрии были исследованы пероксидация липидов, общий глутатион и активность ферментов. Плазма была также использована для определения концентрации гемоглобина как показателя гемолиза. Результаты. Выполнение максимального аэробного теста спровоцировало значительное повышения уровня гемоглобина в плазме крови и снижение пероксидации липидов, однако никак не повлияло на уровень общего глутатиона в эритроцитах. Что касается антиоксидантных ферментов, то результаты анализа показали увеличение активности глутатионпероксидазы, при этом активность каталазы и супероксиддисмутазы осталась на том же уровне. Заключение. Можно прийти к выводу, что у активных спортсменов преобладают эритроциты, которые более устойчивы к оксидативному стрессу вследствие ускоренного гемолиза, спровоцированного физическими упражнениями, которые приводят к удалению старых и оксидативно модифицированных клеток.

This study aimed to establish the sensitivity of Muscovy duck semen to oxidative stress (OS) and the effect of Desferal, applied as an antioxidant. The effect of three prooxidant systems in presence and absence of Desferal were tested on the motility and kinetic parameters (determined using CASA system), as well as the level of lipid peroxidation (LPO) and glutathione (tGSH) of Muscovy semen. The semen was diluted (1:3 v/v) with four extenders (saline solution, IMV Canadyl, HIA-1, and AU) and stored at 4 °C for 6 h. The cooled semen was divided into aliquots (50 × 10

Oxidative stress underlies the pathogenesis of many diseases, which determines the interest in natural substances with antioxidant properties. Ginkgo biloba L. leaves are well known and widely used in the pharmaceutical industry, but the therapeutic properties of the seeds are less studied. This study aimed to identify the chromatographic profile and to evaluate the antioxidant properties of methanol extract from seeds of G. biloba (GBSE). In the GBSE, flavonoids and terpenes were found as terpenes predominated. The GBSE antioxidant capacity determined by 2,2 azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) and 1-diphenyl-2-picrylhydrazyl (DPPH) methods were equal to 1.34% and 0.58% of the activity of reference substance Trolox, respectively. The results of the ferric reducing antioxidant power method showed that the effect of concentration 1 mg/mL (w/v) GBSE was equal to 7.418 mM FeSO4 used as a standard. The cupric reducing antioxidant capacity activity of the GBSE was found to be 215.39 µmol Trolox/g GBSE and is presented as Trolox equivalent. The metal chelation effect of 1 mg/mL (w/v) GBSE was equal to that obtained for 0.018 mM EDTA. In conclusion, GBSE showed a good ability to neutralize ABTS and DPPH radicals and could have a beneficial effect in pathological conditions with oxidative stress etiology.

Plants from the Rosacea family are rich in natural molecules with beneficial biological properties, and they are widely appreciated and used in the food industry, perfumery, and cosmetics. In this review, we are considering Rosa damascena Mill., Rosa alba L., Rosa centifolia L., and Rosa gallica L. as raw materials important for producing commercial products, analyzing and comparing the main biological activities of their essential oils, hydrolates, and extracts. A literature search was performed to find materials describing (i) botanical characteristics; (ii) the phytochemical profile; and (iii) biological properties of the essential oil sand extracts of these so called “old roses” that are cultivated in Bulgaria, Turkey, India, and the Middle East. The information used is from databases PubMed, Science Direct, and Google Scholar. Roses have beneficial healing properties due to their richness of beneficial components, the secondary metabolites as flavonoids (e.g., flavones, flavonols, anthocyanins), fragrant components (essential oils, e.g., monoterpenes, sesquiterpenes), and hydrolysable and condensed tannins. Rose essential oils and extracts with their therapeutic properties—as respiratory antiseptics, anti-inflammatories, mucolytics, expectorants, decongestants, and antioxidants—are able to act as symptomatic prophylactics and drugs, and in this way alleviate dramatic sufferings during severe diseases.

Allium siculum Ucria is a perennial bulbous plant that occurs in Bulgaria as A. siculum subsp. dioscoridis (Sm.) K. Richt. The plant is locally known as “samardala”. Its herbage is used traditionally by local communities in Bulgaria as edible green and for preparation of salty spice mixtures. The study was focused on the documentation of traditional knowledge about samardala, covering local collection and cultivation practices, processing methods and consumption preferences. The respondents (aged 30–86 years, 50% over 65) were local producers and small-scale salesmen of the herbage and samardala-containing spices. To assess the health features of samardala spices the total phenol and flavonoid content was measured in home-consumed or marketed flavoured salts. Overall appreciation of the plant and related food products was found to be very high; presenting a considerable engagement of the locals in plant cultivation and gathering, laborious processing and consumption. Traditional ecological knowledge about the biology, phenology and ecology of the samardala plant and methods for its processing is still preserved by elderly people who rely mostly on their own cultivated plants. Although the introduction of modern appliances has altered the method of production, its main stages and the recipes are preserved. Modernization of the processing method has not affected the quality of the products and faster processing could be a contributing factor for the higher content of biologically active substances. Phenolic content was about twice as low as the flavonoid content, following the same pattern for all tested flavoured salts. Measures for better management of natural populations and promotion of cultivation practices of A. siculum subsp. dioscoridis would ensure the sustainable quality of traditional products as well as conservation of the natural plant resources.

The wastewater from the distillation of rose oils is discharged directly into the soil because it has a limited potential for future applications. The aim of the present study was to determine in vitro the chromatographic profile, redox-modulating capacity, and antineoplastic activity of wastewater obtained by distillation of essential oils from the Bulgarian Rosa alba L., Rosa damascena Mill., Rosa gallica L., and Rosa centifolia L. We applied UHPLC-HRMS for chromatographic analysis of rose wastewaters, studied their metal-chelating and Fe(III)-reducing ability, and performed MTT assay for the evaluation of cytotoxic potential against three tumorigenic (HEPG2—hepatocellular adenocarcinoma, A-375—malignant melanoma, A-431—non-melanoma epidermoid squamous skin carcinoma) and one non-tumorigenic human cell lines (HaCaT—immortalized keratinocytes). The median inhibitory concentrations (IC50) were calculated with nonlinear modeling using the MAPLE® platform. The potential of the wastewaters to induce apoptosis was also examined. Mono-, di-, and acylated glycosides of quercetin and kaempferol, ellagic acid and its derivatives as main chemical components, and gallic acid and its derivatives—such as catechin and epicatechin—were identified. The redox-modulating capacity of the samples (TPTZ test) showed that all four wastewaters exhibited the properties of excellent heavy metal cleaners, but did not exert very strong cytotoxic effects. The lowest IC50 rate was provided in wastewater from R. centifolia (34–35 µg/mL of gallic acid equivalents after a 72 h period for all cell lines). At 24 and 48 hours, the most resistant cell line was HEPG2, followed by HaCaT. After 72 h of exposure, the IC50 values were similar for tumor and normal cells. Still, R. damascena had a selectivity index over 2.0 regarding A-431 non-melanoma skin cancer cells, showing a good toxicological safety profile in addition to moderate activity—IC50 of 35 µg/mL polyphenols. The obtained results related to wastewaters acquired after the distillation of essential oils from the Bulgarian R. alba, R. damascena, R. gallica, and R. centifolia direct our attention to further studies for in-depth elucidation of their application as detoxifying agents under oxidative damage conditions in other experimental datasets.

The pathogenesis of many diseases and different pathological conditions, including inflammation, is associated with excess production of reactive oxygen species (ROS). The present study aimed to investigate the effects of the antidepressant desipramine (DES) on carrageenan (CG)-induced inflammation, as well as on the endogenous levels of cell enzyme and non-enzyme antioxidants in rat liver and spleen, 4 and 24 h after CG injection. The intra-plantar CG injection into the right hind paw resulted in a time-dependent increase in the paw volume; the maximum of CG-induced edema peak was in 2–4 h. A single DES dose of 20 mg·kg−1, administered 30 min before CG, had no effect on paw edema, whereas the higher drug dose used (50 mg·kg−1) suppressed the edematous response to CG. The latter drug dose protected CG-induced decrease of glutathione (non-enzyme antioxidant) in the liver; it did not affect CG-unchanged activities of superoxide dismutase, glutathione peroxidase (enzyme antioxidants) and glucose-6-phosphate dehydrogenase (enzyme, important for the activity of glutathione-conjugated antioxidant enzymes) in both liver and spleen. The drug showed an efficient antioxidant capacity in ROS-generating chemical systems; it was higher than that of fluoxetine (another type of antidepressant). The present results suggest that the good antioxidant activity of DES might contribute to its beneficial effects in liver injuries. Copyright © 2011 John Wiley & Sons, Ltd.

The in vivo effects of nociceptin (N/OFQ(1-13)NH(2) ) and its structural analogues ([Dab(9) ]N/OFQ(1-13)NH(2) , [Dap(9) ]N/OFQ(1-13)NH(2) and [Cav(9) ]N/OFQ(1-13)NH(2) ) on the levels of lipid peroxidation and cell antioxidants (enzyme and non-enzyme) in brain of control and kainic acid (KA)-treated rats were studied. In control animals, [Dab(9) ]N/OFQ(1-13)NH(2) and [Dap(9) ]N/OFQ(1-13)NH(2) , unlike N/OFQ(1-13)NH(2) and [Cav(9) ]N/OFQ(1-13)NH(2) , slightly increased the brain lipid peroxidation; the rest of the parameters were unchanged by all neuropeptides tested. KA (0.25 µg in 0.5 µl, i.c.v) increased the lipid peroxidation (4 and 24 h after KA-injection) and decreased the glutathione level (1 h after KA-administration). One hour after KA-administration, the neuropeptides (2 µg in 0.5 µl, injected 30 min before KA) showed the following effects: a slight decrease in the KA-induced lipid peroxidation by all nociceptin analogues and an enhancement of the KA-decreased GSH level, but by [Cav(9) ]N/OFQ(1-13)NH(2) only. The brain antioxidant enzyme activities were unchanged in all used experimental groups. In addition, the nociceptin analogues, especially [Can(9) ]N/OFQ(1-13)NH(2) , showed a good antioxidant capacity in chemical systems, generating reactive oxygen species. In conclusion, the substitution of lysin (Lys) in N/OFQ(1-13)NH(2) molecule with other amino acids might contribute to changes in its antioxidant properties. Copyright © 2011 John Wiley & Sons, Ltd.

Aim: Previous studies have shown that proteasome inhibitors exerted protective effects against ischemia/reperfusion injury (IRI) of brain, heart, kidney and intestine. The aim of the present study was to investigate: (i) whether the proteasome inhibitor MG132 protects rat liver against IRI; and (ii) whether MG132 modulates prooxidant/antioxidant status of rat liver subjected to warm IRI. Methods: The left lateral and medial lobes (approximately 70% of the total liver volume) of livers of male Wistar rats were subjected to 30-min ischemia followed by 60-min reperfusion. Lactate dehydrogenase (LDH), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) levels were measured in the plasma. Proteasome chymotryptic-like (ChT-L) activity, levels of thiobarbituric acid-reactive substances (TBARS), protein carbonyls (PC) and glutathione (GSH), as well as superoxidase dismutase (SOD), catalase (CAT), glutathionine peroxidase and glutathionine reductase activities were measured in liver fractions. Results: Thirty-min ischemia followed by 60-min reperfusion increased liver TBARS and PC, CAT and SOD activities, but decreased GSH level. Ischemia/reperfusion-induced oxidative stress was exacerbated in mitochondria, indicating that these organelles are the preferential target of IRI. Plasma LDH and AST levels were decreased by MG132 during both ischemia and reperfusion, while ALT values were decreased only after 30 min of reperfusion. MG132 did not significantly affect liver TBARS and GSH levels, but it increased PC and decreased ChT-L activity; the activities of CAT and SOD were also decreased. Conclusions: MG132 exerts a protective effect during the early phase of reperfusion and it modulates prooxidant/antioxidant status of rat liver subjected to warm IRI.

In vivo effects of N-benzyloxycarbonyl (Cbz)-Leu–Leu-leucinal (MG132) on chymotryptic-like (ChT-L), tryptic-like, and post-glutamyl peptide hydrolytic-like proteasome activities, protein oxidation, lipid peroxidation (LP), glutathione (GSH) level, as well as on the activity of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione-reductase) in the rat liver were studied. The possibility of MG132 provoking the formation of free oxygen radicals was also assayed in primary hepatocytes. The following results were obtained: (1) In vivo, MG132 did not change the spontaneous LP, but increased Fe-induced LP and the amount of oxidized proteins; it decreased the GSH level in liver. From the proteasome activities studied in liver cytosol only ChT-L activity was significantly decreased after MG132 administration. Furthermore, MG132 increased antioxidant enzyme activities of SOD, CAT, and GSH-Px. (2) In vitro, MG132 increased free radical oxygen species in hepatocytes; this effect disappeared in the presence of CAT or mannitol. In conclusion, since nowadays proteasome inhibitors are entering into the swing of laboratory and clinical practice, the present data could provide useful information for MG132 action. Consequently, future in vivo experiments with MG132 could highlight the possibility of its use at different pathological conditions. Copyright © 2008 John Wiley & Sons, Ltd.

Summary 1 In vitro studies of the potential antioxidant activity of the selective cyclo-oxygenase-2 inhibitor celecoxib and the non-steroid anti-inflammatory drug amtolmetin guacyl (AMG) were carried out. The study included experiments on the ability of these drugs to affect some indices of the oxidative stress [lipid peroxidation (LP), activity of antioxidant enzymes, glutathione (GSH) level] in rat stomach and colon mucosa and in liver. 2 Celecoxib and AMG did not change the activity of the enzymes GSH-peroxidase, oxidased glutathione (GSSG)-reductase and glucose-6-phosphate-dehydrogenase, as well as the GSH level in all tissue preparations. An increased superoxide dismutase (SOD) activity and a tendency to a decreased Fe/ascorbic acid-induced LP in stomach and colon mucosa were found, but only in the presence of AMG. 3 In the liver, both celecoxib and AMG decreased spontaneous and Fe/ascorbic acid-induced LP. SOD activity was enhanced only in the presence of AMG. 4 Experiments aimed at studying celecoxib and AMG in free oxygen radical-generating systems were also carried out. AMG and tolmetin (the main metabolite of AMG) inhibited OH•-provoked deoxyribose degradation in a Fenton system. Celecoxib had no effect on free radicals when tested in the same system. 5 In conclusion, the results of the present in vitro studies suggest that AMG and celecoxib possess antioxidant and metal-chelating abilities, which might contribute to their beneficial effects.

The pathogenesis of many diseases and different pathological conditions, including inflammation, is associated with excess production of reactive oxygen species (ROS). The present study aimed to investigate the effects of the antidepressant desipramine (DES) on carrageenan (CG)-induced inflammation, as well as on the endogenous levels of cell enzyme and non-enzyme antioxidants in rat liver and spleen, 4 and 24 h after CG injection. The intra-plantar CG injection into the right hind paw resulted in a time-dependent increase in the paw volume; the maximum of CG-induced edema peak was in 2-4 h. A single DES dose of 20 mg · kg(-1) , administered 30 min before CG, had no effect on paw edema, whereas the higher drug dose used (50 mg · kg(-1) ) suppressed the edematous response to CG. The latter drug dose protected CG-induced decrease of glutathione (non-enzyme antioxidant) in the liver; it did not affect CG-unchanged activities of superoxide dismutase, glutathione peroxidase (enzyme antioxidants) and glucose-6-phosphate dehydrogenase (enzyme, important for the activity of glutathione-conjugated antioxidant enzymes) in both liver and spleen. The drug showed an efficient antioxidant capacity in ROS-generating chemical systems; it was higher than that of fluoxetine (another type of antidepressant). The present results suggest that the good antioxidant activity of DES might contribute to its beneficial effects in liver injuries.

The present study aimed to investigate the effects of pentoxifylline (PTX) on the carrageenan (CG)-induced paw oedema and on the endogenous levels of cell enzyme and non-enzyme antioxidants in rat liver, 4 and 24 h after CG injection. PTX (50 mg kg(-1) , i.p.), administered 30 min before CG, decreased the paw oedema, 2-4 h after CG administration. The drug protected CG-induced decrease of glutathione (non-enzyme antioxidant) and had no effect on CG-unchanged activities of superoxide dismutase, glutathione peroxidase (enzyme antioxidants) and glucose-6-phosphate dehydrogenase (enzyme, important for the activity of GSH-conjugated antioxidant enzymes). The drug showed a good antioxidant capacity in chemical systems, generating reactive oxygen species. The present results suggest that the antioxidant activity of PTX might contribute to its beneficial effects in liver injuries.

The in vivo effects of nociceptin (N/OFQ(1-13)NH(2) ) and its structural analogues ([Dab(9) ]N/OFQ(1-13)NH(2) , [Dap(9) ]N/OFQ(1-13)NH(2) and [Cav(9) ]N/OFQ(1-13)NH(2) ) on the levels of lipid peroxidation and cell antioxidants (enzyme and non-enzyme) in brain of control and kainic acid (KA)-treated rats were studied. In control animals, [Dab(9) ]N/OFQ(1-13)NH(2) and [Dap(9) ]N/OFQ(1-13)NH(2) , unlike N/OFQ(1-13)NH(2) and [Cav(9) ]N/OFQ(1-13)NH(2) , slightly increased the brain lipid peroxidation; the rest of the parameters were unchanged by all neuropeptides tested. KA (0.25 µg in 0.5 µl, i.c.v) increased the lipid peroxidation (4 and 24 h after KA-injection) and decreased the glutathione level (1 h after KA-administration). One hour after KA-administration, the neuropeptides (2 µg in 0.5 µl, injected 30 min before KA) showed the following effects: a slight decrease in the KA-induced lipid peroxidation by all nociceptin analogues and an enhancement of the KA-decreased GSH level, but by [Cav(9) ]N/OFQ(1-13)NH(2) only. The brain antioxidant enzyme activities were unchanged in all used experimental groups. In addition, the nociceptin analogues, especially [Can(9) ]N/OFQ(1-13)NH(2) , showed a good antioxidant capacity in chemical systems, generating reactive oxygen species. In conclusion, the substitution of lysin (Lys) in N/OFQ(1-13)NH(2) molecule with other amino acids might contribute to changes in its antioxidant properties. Copyright © 2011 John WileySons, Ltd.

Copper toxicity is associated with formation of reactive oxygen species, which are capable to oxidize proteins. The selective removal of the latter by the 20S proteasome is considered an essential part of the cell antioxidant defense system. The aim of the present study was to investigate whether peptidase activities of rat liver proteasomes were affected by chronic (40 mg CuSO(4)/rat/daily with the drinking water for 2 weeks) and acute (20 mg/kg CuSO(4), s.c.) copper treatment. To evaluate the role of proteasome, its inhibitor MG132 was also used. The degree of copper-induced oxidative stress (OS), established by measuring lipid peroxidation, protein oxidation, and cellular glutathione level, as well as activities of antioxidant enzymes--catalase, superoxide dismutase, and gultathionine peroxidase, depended on the mode of copper administration. Chronic copper administration (mild oxidative stress) did not affect proteasome activities, whereas acute copper treatment (severe oxidative stress) caused a decline in chymotryptic- and tryptic-like activities. The treatment of copper-loaded animals with MG132 did not change copper-induced alterations in the tested indices, except an additional increase in protein oxidation and inhibition of glutathionine peroxidase activity. The results suggested that the in vivo copper-induced oxidative stress was associated with changes in the catalytic activity of proteasome.

The in vitro effects of arachidonyl-2-chloroethylamide (ACEA; a selective CB(1)-receptor agonist) and N-piperidin-l-yl)-5-(4-chlorophenyl)-1-(2,4-cochlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (SR141716A; a selective CB(1)-receptor antagonist) on lipid peroxidation (spontaneous and Fe (2+)-induced), total glutathione (GSH)-level and activity of antioxidant enzymes (superoxide dismutase, glutathione peroxidase and glutathione reductase) in the rat brain were studied. The effects of these CB(1)-induced), total glutathione (GSH)-level and activity of antioxidant enzymes (superoxide dismutase, glutathione peroxidase and glutathione reductase) in the rat brain were studied. The effects of these CB(1)-ligands in Fenton system (generating *OH radicals) were also examined. The cannabinoids did not change the total GSH-level and were without effects on the activity of antioxidant enzymes in the rat brain. These results proved a lack of in vitro pro-oxidant activity of the CB(1)-receptor ligands, as well as a lack of direct effects on GSH and enzyme molecules. ACEAand SR141716A were without effect on spontaneous lipid peroxidation, but decreased the Fe(2+)-induced brain lipid peroxidation and OH-provoked deoxyribose degradation in Fenton system. It can be suggested that the tested cannabinoids possess a metal-chelating activity, which might contribute to an antioxidant activity. The data, obtained in this study offer a background for investigation of the in vivo effects of these CB(1)-receptor ligands on antioxidant defense systems in the brain of healthy animals and animals, previously subjected to oxidative stress.