Cardiovascular effects elicited by milonine, a new 8,14-dihydromorphinandienone alkaloid

Abstract: The mechanisms underlying the cardiovascular responses evoked by milonine (i.v.), an alkaloid, were investigated in rats. In normotensive rats, milonine injections produced hypotension and tachycardia, which were attenuated after N^w‐nitro‐l ‐arginine methyl esther (l ‐NAME; 20 mg/kg, i.v.). In phenylephrine (10 μM), pre‐contracted mesenteric artery rings, milonine (10^?10 M to 3 × 10^?4 M) caused a concentration‐dependent relaxation (EC₅₀ = 1.1 × 10^?6 M, E_max = 100 ± 0.0%) and this effect was rightward shifted after either removal of the vascular endothelium (EC₅₀ = 1.6 × 10^?5, p < 0.001), or after l ‐NAME 100 μM (EC₅₀ = 6.2 × 10^?5, p < 0.001), hydroxocobalamin 30 μM (EC₅₀ = 1.1 × 10^?4, p < 0.001) or ODQ 10 μM (EC₅₀ = 1.9 × 10^?4p < 0.001). In addition, in rabbit aortic endothelial cells, milonine increased NO₃^? levels. The relaxant effect induced by milonine was attenuated in the presence of KCl (20 mM), a modulator efflux K⁺ (EC₅₀ = 1.2 × 10^?5, p < 0.001), or different potassium channel blockers such as glibenclamide (10 μM) (EC₅₀ = 6.3 × 10^?5, p < 0.001), TEA (1 mM) (EC₅₀ = 2.3 × 10^?5 M, n = 6) or Charybdotoxin (0.2 μM) plus apamin (0.2 μM) (EC₅₀ = 3.9 × 10^?4 M, n = 7). In addition, pre‐contraction with high extracellular potassium concentration prevented milonine‐induced vasorelaxation (EC₅₀ = 1.0 × 10^?4, p < 0.001). Milonine also reduced CaCl₂‐induced contraction in Ca²⁺‐free solution containing KCl (60 mM). In conclusion, using combined functional and biochemical approaches, we demonstrated that the hypotensive and vasorelaxant effects produced by milonine are, at least in part, mediated by the endothelium, likely via nitric oxide release, activation of nitric oxide‐cGMP pathway and opening of K⁺ channels.     

Earthworm biomarker responses on exposure to commercial cypermethrin

Cypermethrin is a synthetic pyrethroid insecticide used worldwide in agriculture, home pest control, disease vector control, and food safety. It accumulates in soil. Therefore, traces of cypermethrin may frequently appear in vegetables grown in contaminated soil. There is a push now to develop biomarkers as early warning indicators of environmental pollution. In this study, DNA damage (tail DNA%, tail length, and olive tail moment), the micronucleus, neutral red retention (NRR) time, and pinocytic adherence ability of coelomocytes were investigated in Pheretima peguana earthworms exposed to cypermethrin in filter paper tests. The NRR time of earthworm coelomocytes decreased significantly at a concentration of 3.5 × 10^?3 µg · cm^?2 (1/100 LC₅₀) after 48 h exposure, with a highly negative correlation with cypermethrin concentration. Pinocytic adherence ability of coelomocytes also declined significantly at a cypermethrin concentration of 3.5 × 10^?2 µg · cm^?2 (1/10 LC₅₀). The DNA damage to earthworm coelomocytes (tail DNA%, tail length, and olive tail moment) increased considerably at the highest concentration (3.5 × 10^?1 µg · cm^?2) although the correlation between tail DNA% and cypermethrin concentration was low. Thus, physiological biomarkers were more sensitive than the genotoxic effects in earthworms exposed to commercial cypermethrin. Although a suite of earthworm biomarkers could be used to evaluate cypermethrin terrestrial pollution, the NRR test is easier to conduct and a more sensitive indicator. © 2013 Wiley Periodicals, Inc. Environ Toxicol 30: 597–606, 2015.     

Mutagenicity and genotoxicity of PM2.5 issued from an urbano‐industrialized area of Dunkerque (France)

Epidemiological studies have demonstrated the link between chronic exposure to particulate matter (PM), especially particles with an aerodynamic diameter lesser than 2.5 µm (PM_2.5), and lung cancer. Mechanistic investigations focus on the contribution of the various genotoxicants adsorbed onto the particles, and more particularly on polycyclic aromatic hydrocarbons or nitroaromatics. Most of the previous studies dealing with genotoxic and/or mutagenic measurements were performed on organic extracts obtained from PM_2.5 collected in polluted areas. In contrast, we have evaluated genotoxic and mutagenic properties of urbano‐industrial PM_2.5 (PM) collected in Dunkerque (France). Thermally desorbed PM_2.5 (dPM) was also comparatively studied. Suspensions of PM and dPM (5–50 µg per plate) were tested in Salmonella tester strains TA98, TA102 and YG1041 ± S9mix. Significant mutagenicity was observed for PM in YG1041 ± S9 mix. In strain TA102 – S9mix, a slight, but not significant dose–response increase was observed, for both PM and dPM. Genotoxic properties of PM and dPM were evaluated by the measurement of (1) 8‐OHdG in A549 cells and (2) bulky DNA adducts on A549 cells and on human alveolar macrophages (AMs) in primary culture. A dose‐dependant formation of 8‐OHdG adducts was observed on A549 cells for PM and dPM, probably mainly attributed to the core of the particles. Bulky DNA adducts were observed only in AMs after exposure to PM and dPM. In conclusion, using relevant exposure models, suspension of PM_2.5 induces a combination of DNA‐interaction mechanisms, which could contribute to the induction of lung cancer in exposed populations. Copyright © 2010 John Wiley & Sons, Ltd.     

Spectrophotometric determination of penicillins in pure and pharmaceutical formulations using Folin‐Ciocalteu reagent

A new combination of time, temperature, and alkali is described for the spectrophotometric determination of amoxicillin and ampicillin using Folin‐Ciocalteu reagent. The method is based on the development of blue‐coloured product due to the reduction of tungstate and/or molybdate in Folin‐Ciocalteu reagent by amoxicillin and ampicillin in alkaline medium. The chromogenic reaction has λ_max at 720 and 740 nm with molar absorptivity 1.6295 × 10⁴ and 0.1085 × 10⁴ l mol^?1 cm^?1 in the Beer's Law range 2–10 µg mL^?1 and 10–70 µg mL^?1 for amoxicillin and ampicillin, respectively. The method is reproducible, quick, inexpensive, and particularly helpful in determining the drug content in commercial dosage forms. Copyright © 2010 John Wiley & Sons, Ltd.     

Cardiovascular effects induced by N-(4'-dihydro)-piperoylthiomorpholine in normotensive rats

Objectives We have tested the cardiovascular effects of N‐(4′‐dihydro)‐piperoylthiomorpholine (LASSBio 365) on rats using an in‐vivo and in‐vitro approach. Methods LASSBio 365 (0.025, 0.05, 0.1, 0.25, 0.5 or 1 mg/kg, randomly injected) was administered to conscious unrestrained rats and the mean arterial pressure and heart rate were measured. The effects of LASSBio 365 (3 × 10^?6–3 × 10^?4m ) on rat isolated aortic rings with and without endothelium were investigated. Key findings LASSBio 365 induced a dose‐dependent decrease in mean arterial pressure and heart rate (ED50 = 158 ± 53 µg/kg). The effects evoked by LASSBio 365 (0.5 mg/kg) were inhibited by pretreatment with atropine. In anaesthetized rats, electrocardiogram recordings revealed second/third degree sinoatrial and atrioventricular blockade induced by the compound, which were completely inhibited after cardiac muscarinic blockade or cervical bilateral vagotomy. In rat isolated aortic rings, LASSBio 365 (3 × 10^?6–3 × 10^?4m ) was capable of antagonizing the contractile effects induced by phenylephrine (1 µm ) or KCl (80 mm ) (IC50 = 107 ± 6; 92 ± 6 µm , respectively). This effect was not inhibited after removal of the vascular endothelium (IC50 = 84 ± 4; 92 ± 10 µm , respectively). LASSBio 365 (10^?6–10^?4m ) antagonized CaCl₂‐induced contractions in a concentration‐dependent manner. Furthermore, LASSBio 365 (98 µm ) inhibited contractions produced by noradrenaline (1 µm ), but not those induced by caffeine (20 mm ). Conclusions These results suggested that LASSBio 365 produced negative chronotropism and reduced peripheral resistance that were probably due to the stimulation of cardiac muscarinic pathways. Peripheral vasodilation was probably linked to voltage‐dependent Ca²⁺‐channel blockade and/or specific inhibition of Ca²⁺ release from noradrenaline‐sensitive intracellular stores.     

Synthesis and cytotoxic evaluation of some new phthalazinylpiperazine derivatives

A new series of 1,4‐disubstituted phthalazinylpiperazine derivatives 7a–f , 12a–f and 20a–f were designed and synthesized in order to develop potent and selective antitumor agents. The target compounds were screened for their cytotoxic activities against A549, HT‐29 and MDA‐MB‐231 cancer cell lines in vitro. Among them, compounds 7a–f exhibited excellent selectivity for MDA‐MB‐231 with IC₅₀ values ranging from 0.013 µM to 0.079 µM. The most promising compound, 7e (IC₅₀ = 2.19 µM, 2.19 µM, 0.013 µM), was 9.3, 10, and 4.9 × 10³ times more active than vatalanib (IC₅₀ = 20.27 µM, 21.96 µM, 63.90 µM), respectively.     

Anti-tumor activity of new artemisinin-chalcone hybrids

In an attempt to develop potent and selective anti‐tumor agents, three new series of artemisinin–chalcone hybrids 10a – 10g , 11a – 11g and 12a–12h were designed, synthesized and screened for their anti‐tumor activity against five cell lines (HT‐29, A549, MDA‐MB‐231, HeLa and H460) in vitro. Among compounds 10a–g and 11a–11g , most of them displayed enhanced activity and good selectivity toward HT‐29 and HeLa cell lines with IC₅₀ values ranging from 0.12 to 0.85 µM as compared with DHA (dihydroartemisinin). Compounds 10a and 11a are most active toward HeLa cells with IC₅₀ values of 0.12 and 0.19 µM. The results revealed that the presence of chalcone moiety is beneficial to their activity and selectivity. In addition, compounds 12a ‐ 12h containing a ‘reversed chalcone’ moiety showed only slight improvement in activity than those of DHA.     

Endoluminal Norepinephrine Inhibits Smooth Muscle Activity of the Pig Pyeloureter by Stimulation of β‐Adrenoceptors without Side Effects

Abstract: It has been demonstrated in pigs that endoluminal administration of norepinephrine reduces the increase in renal pelvic pressure during perfusion. The purposes were to describe concentration–response relationship and receptor mechanism of the effect of norepinephrine on muscle function of pyeloureter and to reveal possible side effects on cardiovascular and renal functions. Renal pelvis was perfused, while pelvic pressure, cardiovascular and renal functional parameters were recorded. In group A, a pelvic pressure increase was examined during pressure flow studies with norepinephrine solutions (0, 1, 5, 50 and 100 µg/ml). In group B, pelvis was perfused with 6 ml/min. norepinephrine solutions (0, 0.001, 0.01, 0.1 and 1 µg/ml). In group C, pelvis was perfused with 6 ml/min. norepinephrine, norepinephrine + sotalol 10^?6 mol/l and norepinephrine + phentolamine 10^?6 mol/l. Norepinephrine solutions of 0, 10^?8, 10^?7, 10^?6, 10^?5 and 10^?4 mol/l were used. In group A, all norepinephrine solutions lowered the pelvic pressure increase significantly. Large increases in plasma and urine norepinephrine occurred with 50 and 100 µg/ml, but cardiovascular and renal functions remained unchanged. In group B, a significant diminishing pelvic pressure increase with all solutions was seen with a significant difference between all solutions. In group C, norepinephrine demonstrated a concentration–response curve with EC₅₀ between 10^?8 and 10^?7 mol/l (10^?7.27±0.40). Sotalol had a smooth muscle inhibitory effect on the pyeloureter and inhibited the effect of norepinephrine increasing EC₅₀ by about a factor 10 (10^?6.40±1.17). No convincing effect of phentolamine was observed. Endoluminal norepinephrine probably stimulates β‐adrenoceptors and inhibits a renal pelvis pressure increase to perfusion in a dose‐related way without side effects. Endoluminal norepinephrine is safe in pigs and may be useful under endoscopy of the pyeloureter.     

Comparative assessment of herbicide phytotoxicity to Selenastrum capricornutum using microplate and flask bioassay procedures

Four reference toxicants (Cr⁶⁺, Cu²⁺, Zn²⁺, phenol) and 9 herbicides (imazamethabenz, 2,4-dichlorophenoxyacetic acid, picloram, glyphosate, bromoxynil, metolachlor, diquat dibromide, hexazinone, cyanazine) were appraised using both the microplate and flask assay Ninety-six hour EC₅₀s determined with Selenastrum capricornutum as the indicator species essentially demonstrated good intermethodological data concordance for all chemicals, with the exception of diquat dibromide, whose phytotoxicity in the microplate assay (EC₅₀ = 4.9 μg · L^?1) was nearly 7 times that of the flask assay (EC₅₀ = 34.2 μg · L^?1). Comparisons with other data in the scientific literature relating to similar herbicides with the same or different green algal indicator species appeared to corroborate the overall data obtained in our study. More than 4 orders of magnitude separated the most toxic (cyanazine, flask, and microplate EC₅₀s of 17.6 and 16.9 μg · L^?1, respectively) and the least toxic (imazamethabenz, flask, and microplate EC₅₀s of 89.1 and 91.1 mg · L^?1, respectively) herbicides. The biprocedural phytotoxicity comparison described in this work suggests that the simpler algal microplate assay can be an appropriate alternative to the flask technique to evaluate the algal growth inhibition effects of herbicides.     

Antimicrobial agent triclosan is a proton ionophore uncoupler of mitochondria in living rat and human mast cells and in primary human keratinocytes

Triclosan (TCS) is an antimicrobial used widely in hospitals and personal care products, at ~10 mm . Human skin efficiently absorbs TCS. Mast cells are ubiquitous key players both in physiological processes and in disease, including asthma, cancer and autism. We previously showed that non‐cytotoxic levels of TCS inhibit degranulation, the release of histamine and other mediators, from rat basophilic leukemia mast cells (RBL‐2H3), and in this study, we replicate this finding in human mast cells (HMC‐1.2). Our investigation into the molecular mechanisms underlying this effect led to the discovery that TCS disrupts adenosine triphosphate (ATP) production in RBL‐2H3 cells in glucose‐free, galactose‐containing media (95% confidence interval EC₅₀ = 7.5–9.7 µm ), without causing cytotoxicity. Using these same glucose‐free conditions, 15 µm TCS dampens RBL‐2H3 degranulation by 40%. The same ATP disruption was found with human HMC‐1.2 cells (EC₅₀ 4.2–13.7 µm ), NIH‐3 T3 mouse fibroblasts (EC₅₀ 4.8–7.4 µm ) and primary human keratinocytes (EC₅₀ 3.0–4.1 µm ) all with no cytotoxicity. TCS increases oxygen consumption rate in RBL‐2H3 cells. Known mitochondrial uncouplers (e.g., carbonyl cyanide 3‐chlorophenylhydrazone) previously were found to inhibit mast cell function. TCS‐methyl, which has a methyl group in place of the TCS ionizable proton, affects neither degranulation nor ATP production at non‐cytotoxic doses. Thus, the effects of TCS on mast cell function are due to its proton ionophore structure. In addition, 5 µm TCS inhibits thapsigargin‐stimulated degranulation of RBL‐2H3 cells: further evidence that TCS disrupts mast cell signaling. Our data indicate that TCS is a mitochondrial uncoupler, and TCS may affect numerous cell types and functions via this mechanism. Copyright © 2015 John Wiley & Sons, Ltd.     

Nano titanium exposure induces dose- and size-dependent cytotoxicity on human epithelial lung and colon cells

The productions as well as use of Titanium dioxide nanoparticles (TNPs) were rapidly increasing in the present nano-world. The TNP becomes an inevitable part our daily life in the form of cosmeceutical, bio-medical, and nano-pharmaceutical applications. The TNPs are either inhaled or ingested into the human body through common routes of exposure like the lungs and the oral-gastrointestinal tract (GIT). Human lung and colon were exposed to test particles, TNP 18?nm (TNP 18), TNP 30?nm (TNP 30), and TNP 87?nm (TNP 87) with a dose range 0.1–100?µg/ml. The effect of exposure was determined using MTT, LDH, and DCFH-DA methods. The TNP 18, TNP 30, and TNP 87 significantly (p?<?0.001) reduced cell viability in a dose- and a size-dependent manner in 60 and 100?µg/ml. The lowest IC₅₀ values 21.80 and 24.83?µg/ml were observed in A549 and Caco-2 for the smallest size, TNP 18. Further, for TNP 30, IC₅₀ values were 23.30 and 28.59?µg/ml compared to Nano QTZ 43.82 and 45.86?µg/ml. The EC₂₅ values of LDH leakage were 5.83 and 9.50?µg/ml for TNP 18 in lung and colon cells. Besides, ROS levels increased significantly at doses 60 (p?<?0.01) and 100 (p?<?0.001) µg/ml in two cells. The smaller size particle, TNP 18 has produced a significant (p?<?0.05) toxic effect at the lowest dose i.e., 10?µg/ml. Therefore, we conclude that TNP 18, TNP 30, and TNP 87 induced a dose- and size-dependent cytotoxicity via decreased cell viability, increased LDH and ROS levels by in vitro methods.     

Investigation on cobalt‐oxide nanoparticles cyto‐genotoxicity and inflammatory response in two types of respiratory cells

The increasing use of cobalt oxide (Co₃O₄) nanoparticles (NPs) in several applications and the suggested genotoxic potential of Co‐oxide highlight the importance of evaluating Co₃O₄ NPs toxicity. Cyto‐genotoxic and inflammatory effects induced by Co₃O₄ NPs were investigated in human alveolar (A549), and bronchial (BEAS‐2B) cells exposed to 1–40 µg ml^–1. The physicochemical properties of tested NPs were analysed by transmission electron microscopy (TEM) and dynamic light scattering (DLS). Cytotoxicity was studied to analyze cell viability (WST1 test) and membrane damage (LDH assay), direct/oxidative DNA damage was assessed by the Formamido‐pyrimidine glycosylase (Fpg)‐modified comet assay and inflammation by interleukin (IL)‐6, IL‐8 and tumor necrosis factor‐alpha (TNF‐α) release (ELISA). In A549 cells, no cytotoxicity was found, whereas BEAS‐2B cells showed a viability reduction at 40 µg ml^–1 and early membrane damage at 1, 5 and 40 µg ml–1. In A549 cells, direct and oxidative DNA damage at 20 and 40 µg ml^–1 were detected without any effects on cytokine release. In BEAS‐2B cells, significant direct DNA damage at 40 µg ml^–1 and significant oxidative DNA damage with a peak at 5 µg ml^–1, that was associated with increased TNF‐α release at 1 µg ml^–1 after 2 h and increased IL‐8 release at 20 µg ml^–1 after 24 h, were detected. The findings show in the transformed alveolar cells no cytotoxicity and genotoxic/oxidative effects at 20 and 40 µg ml^–1. In normal bronchial cells, moderate cytotoxicity, direct DNA damage only at the highest concentration and significant oxidative‐inflammatory effects at lower concentrations were detected. The findings confirm the genotoxic‐oxidative potential of Co₃O₄ NPs and show greater sensitivity of BEAS‐2B cells to cytotoxic and oxidative‐inflammatory effects suggesting the use of different cell lines and multiple end‐points to elucidate Co₃O₄ NPs toxicity. Copyright © 2015 John Wiley & Sons, Ltd.     

Potent enhancement of transdermal absorption and stability of human tyrosinase plasmid (pAH7/Tyr) by Tat peptide and an entrapment in elastic cationic niosomes

Abstract

Enhancement of transdermal absorption through rat skin and stability of the human tyrosinase plasmid (P) using Tat (T) and an entrapment in elastic cationic niosomes (E) were described. E (Tween61:cholesterol:DDAB at 1:1:0.5 molar ratio) were prepared by the freeze-dried empty liposomes (FDELs) method using 25% ethanol. TP was prepared by a simple mixing method. TPE was prepared by loading T and P in E at the T:P:E ratio of 0.5:1:160 w/w/w. For gel formulations, P, TP, PE and TPE were incorporated into Carbopol 980 gel (30?µg of plasmid per 1?g of gel). For the transdermal absorption studies, the highest cumulative amounts and fluxes of the plasmid in viable epidermis and dermis (VED) were observed from the TPE of 0.31?±?0.04?µg/cm² and 1.86?±?0.24?µg/cm²/h (TPE solution); and 4.29?±?0.40?µg/cm² and 25.73?±?2.40?µg/cm²/h (TPE gel), respectively. Only plasmid from the PE and TPE could be found in the receiving solution with the cumulative amounts and fluxes at 6?h of 0.07?±?0.01?µg/cm² and 0.40?±?0.08?µg/cm²/h (PE solution); 0.10?±?0.01?µg/cm² and 0.60?±?0.06?µg/cm²/h (TPE solution); 0.88?±?0.03?µg/cm² and 5.30?±?0.15?µg/cm²/h (PE gel); and 1.02?±?0.05?µg/cm² and 6.13?±?0.28?µg/cm²/h (TPE gel), respectively. In stability studies, the plasmid still remained at 4?±?2?°C and 25?±?2?°C of about 48.00–65.20% and 27.40–51.10% (solution); and 12.34–38.31% and 8.63–36.10% (gel), respectively, whereas at 45?±?2?°C, almost all the plasmid was degraded. These studies indicated the high potential application of Tat and an entrapment in elastic cationic niosomes for the development of transdermal gene delivery system.     

Particle uptake efficiency is significantly affected by type of capping agent and cell line

Surface‐functionalized silver nanoparticles (AgNPs) are the most deployed engineered nanomaterials in consumer products because of their optical, antibacterial and electrical properties. Almost all engineered nanoparticles are coated with application‐specific capping agents (i.e. organic/inorganic ligands on particle surface) to enhance their stability in suspension or increase their biocompatibility for biomedicine. The aim of this study was to investigate the contribution of the selected capping agents to their observed health impacts using realistic dose ranges. AgNPs capped with citrate, polyvinylpyrrolidone (PVP) and tannic acid were studied with human bronchoalveolar carcinoma (A549) and human colon adenocarcinoma (Caco‐2) cell lines and compared against exposures to Ag ions. Cellular uptake and cytotoxicity were evaluated up to 24 h. Tannic acid capped AgNPs induced higher cellular uptake and rate in both cell lines. Citrate‐capped and PVP‐capped AgNPs behaved similarly over 24 h. All three of the capped AgNPs penetrated more into the A549 cells than Caco‐2 cells. In contrast, the uptake rate of Ag ions in Caco‐2 cells (0.11 ± 0.0001 µg h^–1) was higher than A549 cells (0.025 ± 0.00004 µg h^–1). The exposure concentration of 3 mg l^–1 is below the EC₅₀ value for all of the AgNPs; therefore, little cytotoxicity was observed in any experiment conducted herein. Exposure of Ag ions, however, interrupted cell membrane integrity and cell proliferation (up to 70% lysed after 24 h). These findings indicate cellular uptake is dependent on capping agent, and when controlled to realistic exposure concentrations, cellular function is not significantly affected by AgNP exposure. Copyright © 2015 John Wiley & Sons, Ltd.     

Synthesis,Cytotoxicity, and Pro‐Apoptosis Activity of Etodolac Hydrazide Derivatives as Anticancer Agents

Etodolac hydrazide and a novel series of etodolac hydrazide‐hydrazones 3 – 15 and etodolac 4‐thiazolidinones 16 – 26 were synthesized in this study. The structures of the new compounds were determined by spectral (FT‐IR, ¹H NMR, ¹³C NMR, HREI‐MS) methods. Some selected compounds were determined at one dose toward the full panel of 60 human cancer cell lines by the National Cancer Institute (NCI, Bethesda, USA). 2‐(1,8‐Diethyl‐1,3,4,9‐tetrahydropyrano[3,4‐b]indole‐1‐yl)acetic acid[(4‐chlorophenyl)methylene]hydrazide 9 demonstrated the most marked effect on the prostate cancer cell line PC‐3, with 58.24% growth inhibition at 10^?5 M (10 µM). Using the MTT colorimetric method, compound 9 was evaluated in vitro against the prostate cell line PC‐3 and the rat fibroblast cell line L‐929, for cell viability and growth inhibition at different doses. Compound 9 exhibited anticancer activity with an IC₅₀ value of 54 µM (22.842 µg/mL) against the PC‐3 cells and did not display any cytotoxicity toward the L‐929 rat fibroblasts, compared to etodolac. In addition, this compound was evaluated for caspase‐3 and Bcl‐2 activation in the apoptosis pathway, which plays a key role in the treatment of cancer.     

Long‐term exposures to di‐n‐butyl phthalate inhibit body growth and impair gonad development in juvenile Murray rainbowfish (Melanotaenia fluviatilis)

The aim of the present study was to evaluate whether long‐term exposures to environmentally relevant concentrations of di‐n‐butyl phthalate (DnBP) disrupt the reproduction‐based endpoints in juvenile Murray rainbowfish (Melanotaenia fluviatilis). Fish were exposed to 5, 15 or 50 µg l^?1 DnBP for 30, 60 and 90 days each, and the effects on survival, body growth, whole‐body concentrations of sex steroid hormones and gonadal development were investigated. The lowest observed effective concentration to affect the condition factor after 90 days was 5 µg l^?1. Complete feminization of the gonad was noted in fish exposed to 5 µg l^?1 for 90 days and to 15 and 50 µg l^?1 of DnBP for 30 or 60 days. After 90 days of exposure to DnBP, the ovaries were regressed and immature as opposed to the control fish which were in early‐vitellogenic stage. Testes, present only in fish exposed to 5 µg l^?1 of DnBP for 30 or 60 days, were immature in comparison to the control fish that contained testes in the mid‐spermatogenic phase. The E2/11‐KT ratio was significantly higher only after exposures to 5 µg l^?1 DnBP for 90 days and 50 µg l^?1 DnBP for 30 days. Our data suggest that exposures to 5 µg l^?1 DnBP for 30 days did not have profound effects on body growth and gonadal differentiation of fish. However, 30 days of exposure to 15 µg l^?1 could interfere with the gonad development and to 50 µg l^?1 could compromise the hormonal profile of juvenile fish. Copyright © 2014 John Wiley & Sons, Ltd.     

Free radical scavenging actions of three Trifolium species in the protection of blood plasma antioxidant capacity in vitro

Abstract

Context: Three clover [Trifolium L. (Leguminosae)] species were selected on the basis of data from traditional medicine, phytochemical profiles, and agricultural significance.

Objective: The in vitro evaluations of free radical scavenging properties, ferric reducing abilities, and antioxidant effects of extracts from T. pratense L. (crude extract and phenolic fraction), T. pallidum L., and T. scabrum L. (phenolic fractions) were performed.

Materials and methods: Activities of the Trifolium extracts were determined at their final concentrations of 1.5–50?µg/ml. Free radical scavenging properties of methanol extract solutions were estimated by the reduction of DPPH^? and ABTS^? radicals. Measurements of the total antioxidant capacity (TAC) were carried out to assess the antioxidant activities of the extracts in human blood plasma under conditions of oxidative stress, induced by 200?μM peroxynitrite.

Results: The phenolic fraction of T. pratense displayed the strongest ABTS^? and DPPH^? radical scavenging effects (EC₅₀ value of 21.69 and 12.27?µg/ml, respectively). The EC₅₀ value for T. pallidum extract attained 29.77 and 30.06?µg/ml. The two remaining extracts were less potent scavengers (EC₅₀ value higher than 50?µg/ml). Similar differences were obtained during evaluation of the ferric reducing abilities. Analysis of antioxidant properties of the extracts in blood plasma did not provide such evident differences in their actions, however, it indicated that the T. pratense phenolic fraction displayed the strongest effect.

Conclusions: The examined Trifolium extracts partly protected blood plasma and enhanced its non-enzymatic antioxidant defense against harmful action of peroxynitrite in vitro.     

Estimation of absorption of aromatic hydrocarbons diffusing from interior materials in automobile cabins by inhalation toxicokinetic analysis in rats

Aromatic hydrocarbons, as well as aliphatic hydrocarbons, diffusing from interior materials in automotive cabins are the most common compounds contributing to interior air pollution. In this study, the amounts of seven selected aromatic hydrocarbons absorbed by a car driver were estimated by evaluating their inhalation toxicokinetics in rats. Measured amounts of these substances were injected into a closed chamber system containing a rat, and the concentration changes in the chamber were examined. The toxicokinetics of the substances were evaluated on the basis of the concentration–time course using a nonlinear compartment model. The amounts absorbed in humans at actual concentrations in automobile cabins without ventilation were extrapolated from the results obtained from rats. The absorbed amounts estimated for a driver during a 2 h drive were as follows (per 60 kg of human body weight): 30 µg for toluene (interior median concentration, 40 µg m^?3 in our previous study), 10 µg for ethylbenzene (12 µg m^?3), 6 µg for o‐xylene (10 µg m^?3), 8 µg for m‐xylene (11 µg m^?3), 9 µg for p‐xylene (11 µg m^?3), 11 µg for styrene (11 µg m^?3) and 27 µg for 1,2,4‐trimethylbenzene (24 µg m^?3). Similarly, in a cabin where air pollution was marked, the absorbed amount of styrene (654 µg for 2 h in a cabin with an interior maximum concentration of 675 µg m^?3) was estimated to be much higher than those of other substances. This amount (654 µg) was approximately 1.5 times the tolerable daily intake of styrene (7.7 µg kg^?1 per day) recommended by the World Health Organization. Copyright © 2010 John Wiley & Sons, Ltd.     

Anticancer effect of Cenchrus ciliaris L

Cenchrus ciliaris L total alcohol and successive extracts of both aerial and root parts were tested for their anticancer activities against lung (A-549), intestinal (CACO), colon (HCT-116), cervical (Hela), hepatocellular (HepG-2), and breast (MCF-7) (PC3) cell lines and compared with the standard drug vinblastine sulphate. The obtained results exhibited direct cytotoxic effect with variable inhibiting effect on the growth of the listed cell lines comparing to vinblastine sulphate as reference standard drug, these effects showed different IC₅₀ ranged from 11.1?±?0.3 to 267?±?µg/ml.All root extracts showed the best activities against most of the tested cell lines specially HepG-2 (Hepatocellular carcinoma) (9?±?2.1?µg/ml) which was somewhat closely related to the effect of vinblastine sulphate (2.93?±?0.3?µg/ml).The highest anticancer effect of Cenchrus ciliaris L aerial parts and root extracts were recorded on HepG-2 (Hepatocellular carcinoma) their IC₅₀ were 12?±?0.8 & 9?±?2.1 respectively, CACO (colorectal carcinoma) their IC₅₀ were 27.2?±?1.6 & 20.5?±?0.6 respectively, A-549 (Lung carcinoma) their IC₅₀ were 14.5?±?0.7& 11.1?±?0.3 respectively which were better than the standard drug especially in case the anticancer effect on CACO (colorectal carcinoma) and A-549 (Lung carcinoma). Chloroform extracts of both aerial and roots achieved the best anticancer activities on all of the cell lines especially with colorectal (CACO) and Lung carcinoma (A-549). Cenchrus ciliaris could be a promising source of new chemical moieties used to target cancer cells.