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Aim
The current study was designed to investigate the effect of berbamine (BBM) on isoproterenol (ISO) induced changes in cardiac marker enzymes, myocardial oxidative stress, lipid profile and expression of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in male Wistar rats. Rats were pretreated with BBM (25 mg/kg) through intraperitoneal injection for 7 days followed by induction of myocardial infarction (MI) by subcutaneous injection of ISO (85 mg/kg) for last two days. Key findings: In the present study, the histopathological findings of the heart tissue showed that BBM treatment significantly minimized the damage induced by ISO. BBM pretreatment showed a significant decrease in heart weight, serum marker enzymes, lipid peroxidation and significant increase in cardiac endogenous enzymatic and non-enzymatic antioxidants compared to the ISO-treated group. In addition, we observed significantly upregulated eNOS expression and downregulated iNOS expression in BBM pretreated group. Thus, BBM protected the rat's heart from ISO-induced myocardial infarction by its antioxidant, and antilipidemic properties. Significance: The results of the present investigation suggested that BBM efficiently ameliorated the ISO-induced myocardial infarction in rats.

Purpose
Cadmium (Cd) is a classic cumulative nephrotoxicant and literature suggests that its toxicity is associated with oxidative stress and inflammation which contribute to pathologies in various tissues. We sought to investigate whether polyphenols isolated from virgin coconut oil (VCO) would modulate nephrotoxicity and inflammation induced by Cd in rats.
Methods
Rats were administered polyphenols prior to and along with Cd (5 mg/kg, orally) for 7 weeks. Serum markers of renal damage, interleukin-6 (IL-6), C-reactive protein (CRP) and nitric oxide (NO) were evaluated; renal activities of antioxidant enzymes, as well as malondialdehyde (MDA) and reduced glutathione (GSH) content were determined. Histopathologic alterations were evaluated to define kidney damage.
Results
Cadmium exposure induced nephrotoxicity and oxidative stress evident by significantly increased serum levels of creatinine, urea, and uric acid along with remarkable depression in renal activities of antioxidant enzymes and GSH with prominent increase in MDA. Inflammatory markers - IL-6, CRP and NO were significantly increased and confirmed by histopathology. Sub-chronic administration of VCO polyphenols attenuated the Cd-induced biochemical alterations compared to Cd control with remarkably improved histopathological observations.
Conclusion
The findings showed that VCO polyphenol supplementation protects against Cd-induced nephrotoxicity via its antioxidant and anti-inflammatory mechanisms in rats.

Alnus japonica has been used as a traditional oriental medicine for many diseases such as fever, haemorrhage and alcoholism. In this study, A. japonica extracts were evaluated for their in vitro antioxidant potentials and anticancer effects in AGS human gastric carcinoma cell line. The antioxidant properties of A. japonica extracts were evaluated using several biochemical assays, including FRAP (ferric reducing antioxidant power) assay, ABTS (2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)), DPPH (2,2-diphenyl-1-picrylhydrazyl), alkyl and hydroxyl radical scavenging activity assay. Our study showed that ethanol extract of A. japonica (AJE) has a more potent antioxidant activity than its water extract. In addition, AJE extract inhibited the cell growth and induced the cell death by increasing reactive oxygen species (ROS) production in AGS cells. Moreover, AJE extract specifically triggered the apoptosis mediated through the activation of caspase-8, 7, 3, and poly-ADP ribose polymerase (PARP). Thus, these results suggest that AJE extract could be potentially useful as a new promising strategy in the therapy for gastric carcinoma cancer.

It is proposed that the low skin permeation potential of palonosetron could be enhanced by the inclusion of chemical permeation enhancers. The objective of this study is to evaluate the influence of various chemical enhancers on the transdermal permeation of palonosetron. Different drugs in adhesive transdermal patches (F1-F5) were prepared using five pressure sensitive adhesives; Duro-Tak 87-4098, Duro-Tak 87-2074, Duro-Tak 87-900A, Duro-Tak 87-9301 and Duro-Tak 87-2287. Patches prepared using Duro-Tak 87-9301 (F5) was further combined with four well-known chemical enhancers. The influence of permeation enhancers (propylene glycol, diethylene glycol monoethyl ether, Tween 80 and oleic acid) on the transdermal flux was evaluated ex vivo. Release of the drug from fabricated patches was carried out for a period of 6 h. Greater amount of drug (12% w/w) was incorporated in the patches prepared using Duro-Tak 87-9301 (F5). Incorporation of skin permeation enhancers significantly (P < 0.001) improves the transdermal flux of palonosetron. Among the permeation enhancers, propylene glycol (5% w/w) shows highest permeation (53.12 ± 5.62 μg/cm²/h), which is ∼4 folds higher than control. Biphasic drug release was noticed in the prepared patches and the rate of release was relatively high with patch F7. This study reveals that the optimized transdermal system with propylene glycol as permeation enhancers can provide effective therapeutic level of palonosetron.

Obesity would result in increased cardiovascular morbidity including endothelial destruction, and miRNAs are recognized as potent regulators on endothelial function. We therefore explored pivotal miRNAs before and after exercise and dietary intervention in obese adults and examined their potential relationships with selected endothelial function and biomarkers. Obese adults were included in an exercise and dietary intervention training program for 2 months. At the beginning and the end, measurements of anthropometric and metabolic parameters were performed. Flow-mediated dilation, endothelial related biochemicals and circulating miR-214 and miR-126 levels were also determined. Results showed that circulating miR-214 and miR-126 levels were significantly enhanced (P < 0.05) by exercise and dietary intervention along with improved endothelial function. The relationship between relative changes of miR-214 and that of endothelial progenitor cells was significant (r = 0.589, P < 0.05); relative expression of miR-126 was also significantly (r = 0.433, P < 0.05) correlated with endothelial nitric oxide synthase. The intervention lead to upregulation of circulating miR-214 and miR-126 in obesity, and these molecular adaptations are associated with improved endothelial function during the restoring process.

Melanins are polymorphous and multifunctional biopolymers with a relatively high concentration of free radicals. EPR spectroscopy was used to study o-semiquinone free radicals in model eumelanins synthesized from 3,4-dihydroxyphenylalanine (DOPA) and tyrosine in the presence of tyrosinase, and melanins isolated from A-375 and G-361 human melanoma malignum cells exposed to two compounds: 5,7-dimethoxycoumarin (DMC) and valproic acid (VPA). Changes were determined in the concentrations of free radicals in the individual melanins from tumour cells treated with DMC and VPA. A strong decrease in the concentrations of free radicals characterizes melanins isolated from tumour cells treated together with DMC and VPA. Slow spin-lattice relaxation processes were noted in the melanins tested with homogeneous broadened EPR spectra. The EPR technique may be useful not only for the elucidation of free radicals in melanins from A-375 and G-361 cells treated with VPA and DMC but it could also be applied to establish the relationship between melanin type and the malignancy of melanoma malignum.

The aim of this study was to investigate the occurrence of toxic effects in liver tissue of rats treated with α-terpinene. All treatments were intraperitoneally administered at doses of 0.5, 0.75 and 1.0 ml kg^-1 during 10 days. Liver samples were collected and assessed by histopathological analysis, caspases -1, -3, -8 assay, biomarkers of hepatic damage and determination of oxidant/antioxidant status (thiobarbituric acid-reactive substances (TBARS), catalase (CAT), superoxide dismutase (SOD), reactive oxygen species (ROS), glutathione S-transferase (GST) and glutathione peroxidase (GPx)). Additionally, the cytotoxic and genotoxic effects were evaluated by comet assay. An increase was observed on TBARS levels and GPx activity on the hepatic tissue. Instead, CAT and SOD activities decreased in rats treated with a dose of 1.0 ml kg^-1 of α-terpinene. Concomitantly, ROS levels increased and GST levels decreased in rats treated with α-terpinene at doses of 0.5, 0.75 and 1.0 ml kg^-1. Also, there was an increase in frequency of damage, damage index and caspases, while cell viability decreased in rats treated with α-terpinene. Alanine aminotransferase and aspartate aminotransferase increased in rats treated with 1.0 ml kg^-1 of α-terpinene. Therefore, α-terpinene induces oxidative stress, cytotoxic and genotoxic effects in liver tissue involving the caspases activation.

This study sought to characterize the soluble free and bound phenolic compounds from shaddock (Citrus maxima) peels and investigate their effect on 3-hydroxy-methyl-3-glutaryl coenzyme A reductase (HMG-CoA reductase) and glutathione-linked enzymes in colon (Caco-2) cells. The radicals scavenging ability and the protective ability of the phenolic extracts against pro-oxidant induced oxidative damage in Caco-2 cells and rat colon homogenates were also investigated. The free phenolics were extracted with 80% acetone (v/v), while bound phenolics were extracted from the alkaline (NaOH) and acid (HCl) hydrolyzed residue with ethyl acetate. HPLC fingerprinting revealed the presence of gallic acid, catechin, chlorogenic acid, caffeic acid, ellagic acid, epicatechin, rutin, quercitrin and quercetin in the extracts. The results revealed that the extracts inhibited HMG-CoA reductase activity and increased the activities of glutathione reductase and glutathione peroxidase in Caco-2 cells. The extracts inhibited peroxyl radical induced oxidation of membrane lipids in Caco-2 cells and malondialdehyde production in rat colon homogenates. Furthermore, the phenolic extracts scavenged radicals [2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl (OH)] and chelated Fe²⁺ in a concentration-dependent manner. This study showed that shaddock peels could serve as a dietary means or nutraceutical source for protecting the colon from degeneration.

pH-dependent sustained-release Tulsion^® microspheres bearing clarithromycin were prepared using quasi-emulsion solvent diffusion method with thermocoat L 30 D 55 as a release retardant. Both, clarithromycin and thermocoat L 30 D 55, were found to be non-hemolytic during in vitro toxicity assay against human red blood cells. Ratiometric optimization of different solvents using phase diagrams was performed on amount of good solvent, bridging liquid, dispersing liquid and poor solvent. The developed microspheres were evaluated for the recovery (67.27 ± 3.3%), average particle size (52.0 ± 0.46 μm) and encapsulation efficiency (61.0 ± 3.1%). Scanning electron microscopy and transmission electron microscopy revealed that the microspheres were smooth in surface and spherical in shape, respectively. The drug release study was conducted at different pH of GIT and it gave a pH dependent release for clarithromycin. The bioavailability study revealed increased AUC (2 fold) and half-life (1.2 fold) of microspheres as compared to plain drug. The manuscript reported the debut work on thermocoat L 30 D 55 based novel drug delivery system, the polymer is safe to be used, quasi emulsion spherical crystallization technique is a good technique to prepare microspheres, the prepared microspheres provides sustain release profile as well as targeting to colon.

The aim of the presented research was the preparation and in vitro and in vivo evaluation of mucoadhesive oral films containing nystatin. Multivariate data analysis was used to evaluate an innovative approach, in which a combination of different mucoadhesive polymers was employed. The purpose of this was to assess the effects of such a combination on non-woven insoluble carmellose textile as a drug-release modifier in the structure of the film.
It was observed that the mucoadhesive films prepared using polyethylene oxide were more plastic, showed less mechanical resistance and shorter in vitro residence time in comparison with films containing sodium carmellose. The textile used in films containing sodium carmellose significantly prolonged both in vitro and in vivo residence times in rabbits from 50 ± 4 min until 74 ± 4 min and from 48 ± 6 until 80 ± 4 min, respectively. A higher degree of substitution by the acid carboxymethyl group of the textile resulted in slower nystatin dissolution, longer in vitro and in vivo residence times, and higher tensile strength. Textural parameters tensile strength and tensile deformation in conjunction with linear discriminant analysis were able to distinguish the degree of substitution of the textile due to its impact on the studied parameters.

A small molecule EGFR inhibitor, 4-(2-(3-(4-(4-(trifluoromethyl)phenyl)thiazol-2-yl)ureido)vinyl)-1,2-phenylene diacetate ( CIU1) was designed in silico by using caffeic scaffold as core structure. The designed compound showed anti-proliferative action against different solid tumor cell lines, particularly metastatic breast cancer cells. CIU1 inhibited the growth of EGFR-overexpressing MDA-MB-468 triple-negative breast cancer cells and wild-type non-small-cell lung cancer H460 cells with IC₅₀ values of 8.96 μM and 12.98 μM, respectively, these anti-proliferative effects of CIU1 were comparable to gefitinib (a specific EGFR inhibitor) or lapatinib (a dual EGFR and HER2 tyrosine kinase inhibitor). Interestingly CIU1 effectively inhibited the invasive hormone-dependent MCF-7 cancer cells with an IC₅₀ 2.34 μM. The immunoblot analyses revealed that CIU1 induced programmed cell death and suppressed EGFR expression in EGFR-overexpressing breast cancer (MDA-MB468) and lung cancer (PC-9) cells. The findings substantiated our design strategy and demonstrated the potential of CIU1 as new lead for further optimization in the development of anticancer drugs against advanced solid tumors.

The development of new treatments for inflammation continues to be of high interest, since long-acting effect is critical for patients. We investigated whether meloxicam-loaded lipid-core nanocapsules (M-NC) have an anti-inflammatory action superior to a free drug (M-F) on a mouse pleurisy model, by analyzing the time-course of leukocytes migration in the pleural fluid. Male adult Swiss mice were divided into six groups for each time (24; 48 and 72 h) and were pretreated with blank nanocapsules (17 ml/kg) or M-NC (5 mg/kg) or free meloxicam (M-F) (5 mg/kg). After pretreatments, mice received saline (0.9%) or carrageenan (Cg) (1%) into pleural cavity. Four hours after Cg or saline administration, animals were killed, pleural cavity was washed and pleural fluid was collected for the determination of total leukocytes. Cytokines levels, differential leukocyte count and α-1-acid glycoprotein (AGP) levels were determined only at 48 h of pretreatment, which had effect on total leukocyte count. M-NC were effective against the increase in total and differential leukocyte counts and pleural exudate caused by Cg, while M-F had no effect. M-NC had superior effect to M-F against the increase in cytokines and AGP levels induced by Cg. In summary, M-NC had a superior anti-inflammatory effect to free drug in Cg-induced pleurisy, supporting the idea that the inflammatory process in tissues facilitates the vectorization of polymeric nanoparticles.

Succinic acid monoethyl ester (EMS) was recently proposed as an insulinotropic agent for the treatment of non-insulin dependent diabetes mellitus. The present study investigated the effect of EMS and Metformin on dearrangement in glycoprotein levels in the streptozotocin-nicotinamide induced type 2 diabeteic model. Succinic acid monoethyl ester was administered intraperitoneally for 30 days to normal and diabetic rats. The effect of EMS on glucose, insulin, and plasma and tissue glycoproteins were studied. The effect of EMS was compared with Metformin, a reference drug. The levels of glucose, glycosylated haemoglobin and plasma glycoproteins containing hexose, hexosamine and fucose were increased significantly whereas the level of plasma insulin and haemoglobin were decreased significantly in diabetic rats. There was a significant decrease in the level of sialic acid and elevated levels of hexose, hexosamine and fucose in the liver and kidney of streptozotocin-nicotinamide diabetic rats. Administration of EMS to diabetic rats was followed by a decreased level of glucose, glycosylated haemoglobin and plasma glycoproteins. The levels of plasma insulin, haemoglobin and tissue sialic acid were increased whereas the levels of tissue hexose, hexosamine and fucose were near normal. The present study indicates that the EMS possesses a significantly beneficial effect on the glycoprotein moiety in addition to its antidiabetic effect.

Taurine (2-aminoethanesulfonic acid) is an organic acid widely distributed in animal tissues. It is involved in many physiological processes. Thus, it is widely discussed especially due to its antioxidant properties. In this study, we focused on the effect of taurine supplementation on the concentration of antioxidants in blood plasma and erythrocytes of Wistar rats. Taurine was applied in feed mixture in the dosage of 0, 1, 250, 500, 750, 1000, 1500, 2000, 2500, 3000, 3500 and 4000 mg/kg. We monitored both enzymatic and non-enzymatic antioxidants - glutathione peroxidase, glutathione reductase, and superoxide dismutase and reduced/oxidized glutathione and metallothionein. Using three different methods 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), Ferric Reducing Antioxidant Power (FRAP) and free radicals, we determined antioxidant capacity. In addition, we monitored levels of uric acid and glucose. Our results revealed significant changes in both enzymatic and non-enzymatic parameters with the increasing taurine supplementation.

The L-arginine is the substrate of nitric oxide production which is involved in the regulation of apoptosis and inflammatory processes. The effect of L-arginine after fatty streaks formation has not been fully understood in hypercholesterolemic model; therefore the main objective of this study was to investigate the effect of L-arginine after fatty streaks were developed in rabbit's aorta. Eighteen male rabbits were fed 1% cholesterol diet for four weeks. One third of the animals were sacrificed randomly to verify fatty streaks formation in the aorta (phase I). Then the high cholesterol diet was replaced with normal diet, and the remaining animals (n=12) were divided into two groups (phase II); group 1 (n=6): normal diet and group 2 (n=6): normal diet plus L-arginine (3% in drinking water). The experiment was continued for more four weeks. The serum levels of lipids and lipoproteins were increased significantly in phase I (p

There is often an imbalance between the intake of n-6 and n-3 polyunsaturated fatty acids in patients with inflammatory and autoimmune diseases. Oxidative damage and the production of reactive oxygen species by various immune cells follow during the ageing process. Caloric restriction induces a transcriptional response of the genes known to inhibit oxidative stress, tumourigenesis, splicing mRNA and inflammation. On the other hand, calorie malnutrition causes the depression of many immune functions. The accessibility of nutritional factors seems to be an important cause of circannual rhythms. Lowered food intake as an adverse effect of chemotherapy may be why the immune system is altered. Nutrients influence several diseases including diabetes, obesity, inflammatory immune dysfunctions, and neuropathies as well as behavioural characteristics and life span.

Equipment for fast and accurate detection of organophosphate nerve agents is developed and tested. The method is based on the spectrophotometric monitoring of the enzyme activity of butyrylcholinesterase after its contact with air in a special absorption unit (a "scrubber") developed for the purpose. The scrubber was made from a glass tube filled with glass beads (diam. 3 mm) and filled with approx. 5 ml of butyrylcholinesterase in a phosphate buffer of pH 7.4. The air sample was bubbled through this solution for 20 s at a flow rate of 80 l hour^-1. Thereafter 8 μl of the enzyme solution were aspirated into the micro-SIA-LOV analyzer and the activity of the enzymes were evaluated by using Ellman's reagent, i.e. 2.5 mmol l^-1 butyrylthiocholine iodide and 0.25 mmol 5,5'-dithiobis (2-nitrobenzoic acid). The absorbance of the coloured reaction product was measured at 412 nm after the reaction time of 60 s. The residue of the absorption liquid was washed away from the absorber and the system was washed with the enzyme solution prior to next analysis. The contaminated air caused partial inhibition of the enzyme activity of the absorption liquid. The activity of the contaminated sample was compared with the activity of the unaffected enzyme (blank measurement). The analysis was controlled by two PCs. The effect of the concentration of analyte in the absorption liquid on the enzyme activity was tested for 10^-5-10^-9 mol l^-1 sarin. A single analysis (including the absorption step) took

The aim of this study was to investigate the protective effect of naringenin on oxidative stress, on pro-inflammatory cytokines like TGF-β1, IL-1β and on programmed cell death in the testicular damage resulting from streptozotocin (STZ) induced diabetes in rats. Diabetes was induced by a single intraperitoneal injection of STZ (50 mg/kg), and the rats were treated with naringenin (5 mg/kg and 10 mg/kg) administered once a day orally for 10 weeks, starting 3 days after the STZ injection. At the end of the study, all animals were sacrificed. Testis tissue and blood samples were collected for the assessment of sperm parameters, and for biochemical and histopathological analysis. Naringenin treatment significantly decreased the levels of elevated tissue TBARS (thio-barbituric acid) and increased the superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) enzyme activities in the testis tissues. The naringenin-treated rats in the diabetic group showed an improved histological appearance, sperm parameters, and serum testosterone levels, along with a decrement of terminal dUTP nick end-labeling (TUNEL) detected program cell death and a reduced over expression of TGF-β1, IL-1β in Sertoli cells and Leydig cells. These results suggest that naringenin is a food supplement potentially beneficial in reducing testicular damage in diabetic rats by decreasing the oxidative stress related to programmed cell death.

The content of nitrite admixture in preparations of dinitrosyl iron complexes (DNIC) with glutathione synthesized by treatment of aqueous solutions of Fe²⁺ + glutathione with gaseous NO (complex 1) or by mixing solutions of S-nitrosoglutathione (GS-NO) with solutions of Fe²⁺ + glutathione (complex 2) was determined using the Griess method and HPLC as well as from the level of HNO₂ formed upon interaction of gaseous NO with acidified distilled water. In both preparations, DNIC were predominantly represented by the binuclear form (B-DNIC). In complex 1, the appearance of nitrite in DNIC solutions was induced by nitrogen dioxide present in gaseous NO; its interaction with NO gives an adduct, which is further hydrolyzed to nitrite in aqueous solutions. In complex 2, the presence of nitrite admixture could appear in the presence of nitrite non-incorporated into GS-NO synthesized by mixing glutathione and nitrite in acid media. The per cent content of nitrite (with respect to the total content of complex 1) was 6%, whereas in complex 2 it was as low as 0.4%. Such a low level of nitrite contamination in the course of conventional synthesis of DNIC with glutathione does not make any significant contribution to their biomedical (e.g., hypotensive or vasodilator) activity.

To further understanding of the multiple temporal relationships of the physiological process, we monitored simultaneously 25 different variables in individual cows. We used 6 Bruna Italiana non - pregnant and non - lactating cows from the same farm. The animals were housed individually in a 12 m2 box under natural 14/10 light/dark cycle. They were fed twice daily and water was available ad libitum. Locomotor activity and heart rate were recorded continuously. The rectal temperature, respiratory rate and blood samples were recorded every 4 hours for 24 consecutive hours. To describe the periodic phenomenon analytically we applied a trigonometric statistical model according to the single cosinor procedure. Twelve of the 25 variables studied showed a daily rhythm: locomotor activity, rectal temperature, respiratory rate, haemoglobin, glucose, creatinine, urea, total cholesterol, total lipids, non-esterified fatty acid (NEFA), phosphorus and magnesium. Our results contribute to the understanding of the capacity for reaction and adaptation of animals to the environment, and to the improvement in their output by intervention in their environmental circumstances and in the breeding process.

Recently, oxidative stress has been identified as a pivotal pathological factor inducing bone osteoporosis. This phenomenon is responsible for low bone density. It alters bone quality and generates bone fractures. Strontium is found to induce osteoblast activity by stimulating bone formation and reducing bone resorption by restraining osteoclasts. Bioglass (BG) has been used to repair bone defects, and, in combination with strontium (BG-Sr), offers an opportunity to treat this disease. This study investigated the potential role of BG-Sr in improving antioxidant activity and regenerative bone capacity, The effects of both BG-Sr and BG were tested on osteoblast SaOS2 and endothelial EAhy926 cell proliferation in vitro. In vivo, BG-Sr and BG were implanted in the femoral condyles of Wistar rats and compared to that of control groups. Cell proliferation increased significantly by 120% at SaOS2 and 127% at EAhy926. Superoxide Dismutase (SOD), Catalase (CAT) and Glutathione Peroxidase (GPx) were significantly enhanced in BG-Sr treated rats compared to other groups. Moreover, a significant decrease of thiobarbituric acid-reactive substances (TBARs) was observed. The Ca/P ratio increase improved progressive bone mineralization. According to these results, BG-Sr ameliorated cell proliferation and developed an antioxidative defense against ROS. The histological findings highlight the BG-Sr implications in the osteoporosis treatment confirmed by bone construction. The development of BG-Sr as a therapeutic biomaterial protecting against oxidative stress might make an effective choice for application in tissue engineering.

Oxime K203 is a new compound designed to be used as an acetylcholinesterase reactivator for the treatment of intoxication following exposure to tabun and certain pesticides. After intramuscular administration of a therapeutic (23 mg/kg) dose, the time-course of plasma concentrations of K203 in rats was determined by HPLC. Maximum concentrations were reached between 40 and 60 min (16.5±2.1 μg/ml in 40 min and 16.6±2.0 in 60 min, respectively) with the concentration being relatively constant during this period. There was no significant effect on the plasma concentration of thiobarbituric acid reactive substances (TBARS) during the administration of K203, indicating an absence of oxidative stress. Indeed, administration of K203 led to a significant increase in low molecular weight antioxidants which could tentatively be interpreted as representing a beneficial effect.

In the food industry, in the process of creating new agricultural plant products, and in the testing of anti-cancer drugs there is often a need to assay multiple samples of low molecular weight antioxidants, plant samples and foods rich in antioxidants, with minimal additional costs and low degrees of uncertainty. With these demands in mind, we decided to study the fully automated assay of antioxidants using not only automated sample measurements but also automated processing of samples and application of reagents. The automated pipetting system epMotion 5075 and the automated spectrophotometer BS 400 were chosen for the assay purposes. Five methods were introduced for the automation: 2-diphenyl-1-picrylhydrazyl (DPPH) test, ferric reducing antioxidant power (FRAP) method, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) based test, N,N-dimethyl-1,4-diaminobenzene (DMPD) based test and the free radicals method. Samples containing one of the four antioxidants (standard rutin, quercitrin, ferulic and gallic acid) in a range 1-1000 μg/ml were used throughout. All of the tested methods were found suitable for implementation in an automated assay. However, some of them, such as the ABTS test failed to assay all tested antioxidants. The coefficients of determination were also unequal. From the analytical point of view, FRAP methods provided the most reliable results in the automated assay; because of the capacity of the method, approximately 240 samples per hour (one sample per 15 seconds) can be assayed using the automated protocol. We were encouraged by the data received and we expect further interest in the practical performance of such automation. As a mean of testing the robustness of our method, in the next step of our study, oxidative status was assessed in model cell lines derived from prostate cancer (PC-3, PNT1A and 22RV1) that were cultured on ellipticine (0, 0.5, 1, 1.5, 2, 2.5, 5, 7.5, 10, 15 μmol/l) supplemented agar. Antioxidant activity was assessed (DPPH, ABTS, FRAP, DMPD, FR) and calculated on the phenolic antioxidant level (rutin, quercitrin, ferulic and gallic acid), and thus an estimation was formulated of the oxidative stress as a result of the impact of anti-cancer drugs. It can be demonstrated that the new method has wide applicability.

Sarin is a potent inhibitor of acetylcholinesterase (AChE). It is known as an agent of chemical warfare and is one of a number of nerve agents misused for chemical terrorism, e.g. on the Tokyo subway attacks. Though effect of sarin on the cholinergic system is well-known, long-term adverse effects and the role of oxidative stress in sarin toxicity remain unknown. The experiment reported here was carried out on laboratory Wistar rats intramuscularly exposed to 0.5-50% of sarin LD₅₀ for one hour. A complex biochemical examination of plasma samples and an assessment of oxidative stress in the liver, kidney, spleen, cerebellum and frontal lobe were performed after euthanasia of the animals. By means of these biochemical markers, we were able to observe the induction of hyperglycaemia in a dose-dependent manner. Other biochemical markers such as transaminases were influenced in a non-standard manner as sarin probably acted as an inhibitor of these markers. Oxidative stress markers and an assessment of AChE activity showed an unequal impact of sarin on different tissues. Significant inhibition of AChE was found in the cerebellum and frontal lobe. Besides this, alterations in reduced glutathione, ferric reducing antioxidant power (FRAP) and thiobarbituric acid reactive substances (TBARS) were proven. In particular, an accumulation occurred of reduced glutathione in the frontal lobe, whereas depletion of FRAP was found in the kidney and spleen, and a strong increase in TBARS occurred in the spleen in a dose-dependent manner. We infer that sarin extensively influences oxidative homeostasis. Surprisingly, the central nervous system seems to be more resistant than the other organs.

Low molecular weight antioxidants (LMWAs) were assayed by square wave voltammetry (SWV) using screen printed electrodes. Standard antioxidants, i.e. uric acid, ascorbic acid, trolox and glutathione, were assayed in order to estimate the sensitivity and standard redox potentials of individual LMWAs. In another experiment, plasma from Grey Partridges was used as model real samples. Ferric reducing antioxidant power (FRAP) was used as a reference method. Two peaks in plasma samples were found by SWV and correlated to FRAP. The SWV peaks were successfully correlated to FRAP. The practical importance of SWV carried out on screen printed electrodes is discussed.

The present study aimed to investigate the roles of fibrin deposition and protease activated receptor-1 (PAR-1) in renal cytokine/chemokine production and inflammatory cell infiltration in rats of different ages. Acute inflammation was induced by lipopolysaccharide (LPS) in rats which were then treated with tranexamic acid (TA), TA+urokinase (UK) or TA+low-molecular-weight heparin (HP). Fibrin deposition, inflammatory cells and expressions of PAR-1, monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule 1 (ICAM-1) were detected. A reduction in fibrin deposition and PAR-1 expression in the LPS+TA+HP group was associated with decreased infiltration of inflammatory cells and down-regulated expressions of MCP-1 and ICAM-1. In the LPS+TA+UK group, the fibrin deposition, but not the PAR-1 expression, was reduced, However, the infiltration of inflammatory cells decreased and the expressions of MCP-1 and ICAM-1 down-regulated. There were significant differences in the fibrin deposition, infiltration of inflammatory cells and expression of PAR-1, MCP-1 and ICAM-1 between young and old rats undergoing the same treatment. These findings demonstrated that fibrin deposition plays more important roles than PAR-1 dose in cytokine/chemokine production and inflammatory cell infiltration in vivo, and ageing may deteriorate the fibrin deposition-induced production of cytokines/chemokines and infiltration of inflammatory cells.

The series of 4-(2',4'-difluorobiphenylyl)-2-methylbutyric acid (deoxoflobufen, 1) and its four amides and two salts were prepared and tested for anti-inflammatory activity in rats and mice, using as models carrageenan-induced paw oedema, pleuritis, and arachidonic acid-induced ear inflammation, and on leucotriene B₄ production in cells.

Sorption isotherms were estimated for two model organophosphorus pesticides - methamidophos and paraoxon-ethyl - and four typical Central European soils endangered by these pesticides: haplic Chernozems, Cambisols, Luvisols, and Fluvisols. A photometric microplate assay based on the recognition capability of the enzyme acetylcholinesterase toward organophosphorous pesticides was used for the construction of sorption isotherms. The sorption capacity of each soil was then determined and it was found that the soils with the highest content of humic acid (haplic Chernozems and haplic Luvisols) sorbed pesticides most. Pesticides in concentrations over the sorption capacity were easily removed from soil by water.

Natural products with antidiabetic activities provide important sources for the development of new drugs in the treatment of diabetes mellitus. This present work focuses on the antidiabetic activity of a hydro-methanolic (2:3) extract of the sepals of Salmalia malabarica on the blood glucose, the carbohydrade metabolic enzyme, oxidative stress, glycated haemoglobin and transaminase activity in streptozotocin (STZ) induced diabetic rats. Diabetic rats show a significant diminution in the activities of hexokinase, glucose-6-phosphate dehydrogenase and an elevation in the activity of glucose-6-phosphatase in the liver and skeletal muscle. Administration of hydro-methanolic extract of the sepals of Salmalia malabarica to diabetic rats resulted in a significant recovery in the parameters concerned. In the liver and kidney, the activities of catalase (CAT) and peroxidase (Px) were decreased significantly and levels of conjugated diene (CD) and thio-barbituric acid reactive substance (TBARS) were increased significantly in diabetic rats which recovered significantly after administration of hydro-methanolic extract of S. malabarica. Serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) activities which are increased in diabetes were restored by the extract. Glycated haemoglobin (HbA_1C) levels were resettled significantly in the extract treated group compared to the diabetic group. The antidiabetic activity of the extract was supported after a comparison with glibenclamide, a standard antidiabetic drug.

The present study was undertaken to investigate the antihyperammonemic efficacy of the leaf extract of Pongamia pinnata, an indigenous plant used in Ayurvedic Medicine in India (PPEt), on blood ammonia, plasma urea, uric acid, non-protein nitrogen and serum creatinine in control and ammonium chloride induced hyperammonemic rats. The levels of blood ammonia, circulatory urea, uric acid, non-protein nitrogen and creatinine increased significantly in rats treated with ammonium chloride and decreased significantly in rats treated with PPEt and ammonium chloride. There were no significant changes in the body weights of the experimental animals when compared to controls. The antihyperammonemic effect of PPEt could be attributed to (1) its nephroprotective effect by means of detoxifying excess urea and creatinine, (2) its free radical scavenging property, and (3) its antioxidant property. The exact mechanism of antihyperammonemic effect PPEt has still to be investigated and isolation of the active constituents is required.