The food contaminant acetamide is not an in vivo clastogen, aneugen, or mutagen in rodent hematopoietic tissue.

Acetamide (CAS 60-35-5) is classified by IARC as a Group 2B, possible human carcinogen, based on the induction of hepatocellular carcinomas in rats following chronic exposure to high doses. Recently, acetamide was found to be present in a variety of human foods, warranting further investigation. The regulatory body JECFA has previously noted conflicting reports on acetamide's ability to induce micronuclei (MN) in mice in vivo. To better understand the potential in vivo genotoxicity of acetamide, we performed acute MN studies in rats and mice, and a subchronic study in rats, the target species for liver cancer. In the acute exposure, animals were gavaged with water vehicle control, 250, 1000, or 2000 mg/kg acetamide, or the positive control (1 mg/kg mitomycin C). In the subchronic assay, bone marrow of rats gavaged at 1000 mg/kg/day (limit dose) for 28 days was evaluated. Both acute and subchronic exposures showed no change in the ratio of polychromatic to total erythrocytes (P/E) at any dose, nor was there any increase in the incidence of micronucleated polychromatic erythrocytes (MN-PCE). Potential mutagenicity of acetamide was evaluated in male rats gavaged with vehicle control or 1500 mg/kg/day acetamide using the in vivoPig-a gene mutation assay. There was no increase in mutant red blood cells or reticulocytes in acetamide-treated animals. In both acute and sub-chronic studies, elevated blood plasma acetamide in treated animals provided evidence of systemic exposure. We conclude based on this study that acetamide is not clastogenic, aneugenic, or mutagenic in vivo in rodent hematopoietic tissue warranting a formal regulatory re-evaluation.

Transformable Sulfoximine Assisted One-Pot Double Annulation of Vinylic C-H Bonds with Unactivated Alkynes.

The methylphenyl sulfoximine (MPS) directing group (DG) successfully promotes the one-pot double annulation of acrylic acids with alkynes under Ru catalysis, which is unprecedented. Diverse arrays of pyrido-fused-isoquinolinone skeletons are fabricated from acrylamides, creating two C-C and two C-N bonds in a single operation. The unsymmetrical annulation with two distinct alkynes is presented. The recovery of methylphenyl sulfoxide, a precursor of MPS, validates the synthetic adaptability of transformable-DG (TfDG) in C-H activation.

Ruthenium-Catalyzed Hydroarylation and One-Pot Twofold Unsymmetrical C-H Functionalization of Arenes.

A methyl phenyl sulfoximine (MPS) is used as a directing group in the ruthenium-catalyzed intramolecular hydroarylation of alkene-tethered benzoic acid derivatives to afford dihydrobenzofurans and indolines in good to excellent yields. A one-pot, unsymmetrical, twofold C-H functionalization involving intramolecular C-C and intermolecular C-C/C-N bond formations is successfully demonstrated by using a single set of catalytic reaction conditions, which is unprecedented thus far. A novel isoquinolone-bearing dihydrobenzofuran is constructed through an unsymmetrical twofold C-H functionalization.

Ruthenium-catalyzed ortho-C-H mono- and di-imidation of arenes with N-tosyloxyphthalimide.

The Ru(II)-catalyzed imidation of the o-C-H bond in arenes with N-tosyloxyphthalimide is realized with the assistance of a methyl phenylsulfoximine (MPS) directing group. This method is applicable to access the hitherto difficult o-C-H di-imidation products. The sequential C-N and C-C bond formation of o-C-H arenes creates peripherally decorated benzoic acid derivatives. The readily removable MPS-DG and easily modifiable phthaloyl moiety make this strategy synthetically viable for constructing highly functionalized C-N bearing arenes and heteroarenes.

The apoptotic effect of hesperetin on human cervical cancer cells is mediated through cell cycle arrest, death receptor, and mitochondrial pathways.

Hesperetin, a flavonoid from citrus fruits, has several bioactivities such as anti-inflammatory, antihypertensive, antiatherogenic effects. However, studies elucidating the role and the mechanism(s) of action of hesperetin in cervical cancer are sparse. In this study, we investigated the mechanism of the antiproliferative and apoptotic actions exerted by hesperetin on human cervical cancer SiHa cells. The viability of SiHa cells was evaluated using the MTT assay, apoptosis by acridine orange/ethidium bromide, propidium iodide, TUNEL assay, and Annexin V-Cy3, cell cycle distribution and mitochondrial transmembrane potential using flow cytometry, and apoptotic marker genes using quantitative real-time PCR. The treatment of SiHa cells with hesperetin (IC50, 650 µm) showed a marked concentration- and time-dependent inhibition of proliferation and induced the G2/M phase in a dose-dependent manner after 24 h. There was an attenuation of mitochondrial membrane potential with increased expression of caspase-3, caspase-8, caspase-9, p53, Bax, and Fas death receptor and its adaptor protein Fas-associated death domain-containing protein (FADD), indicating the participation of both death receptor- and mitochondria-related mechanisms. Furthermore, hesperetin-induced apoptosis was confirmed by TUNEL and Annexin V-Cy3. This study shows that hesperetin exhibits a potential anticancer activity against human cervical cancer cell lines in vitro through the reduction in cell viability and the induction of apoptosis. Altogether, these data sustain our contention that hesperetin has anticancer properties and merits further investigation as a potential therapeutic agent.

Aluminium oxide nanoparticles induce mitochondrial-mediated oxidative stress and alter the expression of antioxidant enzymes in human mesenchymal stem cells.

An urgent need for toxicological studies on aluminium oxide nanoparticles (Al(2) [Formula: see text]NPs) has arisen from their rapidly emerging range of applications in the food and agricultural sectors. Despite the widespread use of nanoscale aluminium and its composites in the food industry, there is a serious lack of information concerning the biological activities of Al(2) [Formula: see text]NPs (ANPs) and their impact on human health. In this preliminary study, the effects of ANPs on metabolic stress in human mesenchymal stem cells (hMSCs) were analysed. The results showed dose-dependent effects, including cellular toxicity. The mitochondrial membrane potential in the hMSCs decreased with increasing ANP concentrations after 24 h of exposure. The expression levels of oxidative stress-responsive enzymes were monitored by RT-PCR. The expression levels of CYP1A and POR were up-regulated in response to ANPs, and a significant down-regulation in the expression of the antioxidant enzyme SOD was observed. Further, dose-dependent changes in the mRNA levels of GSTM3, GPX and GSR were noted. These findings suggest that the toxicity of ANPs in hMSCs may be mediated through an increase in oxidative stress. The results of this study clearly demonstrate the nanotoxicological effects of ANPs on hMSCs, which will be useful for nanotoxicological indexing.

Naringin induces death receptor and mitochondria-mediated apoptosis in human cervical cancer (SiHa) cells.

Cervical cancer is the second most common female cancer worldwide, and it remains a challenge to manage preinvasive and invasive lesions. Fruit-based cancer prevention entities, such as flavonoid and their derivatives, have demonstrated a marked ability to inhibit preclinical models of epithelial cancer cell growth and tumor formation. Here, we extend the role of naringin-mediated chemoprevention to that of cervical carcinogenesis. The present study sought to investigate the therapeutic potential effect of naringin on apoptosis in human cervical SiHa cancer cells. Viability of SiHa cells was evaluated by the MTT assay, apoptosis and mitochondrial transmembrane potential by flow cytometry, and pro-apoptotic related genes by Real-time quantitative PCR. Naringin showed a 50% inhibition of SiHa human cervical cancer cells at a concentration of 750µM. SiHa cells exhibited apoptotic cell death, intranucleosomal DNA fragmentation, morphological changes and decline in the mitochondrial transmembrane potential. In addition, administration of naringin increased the expression of caspases, p53 and Bax, Fas death receptor and its adaptor protein FADD. These results suggest that the induction of apoptosis by naringin is through both death-receptor and mitochondrial pathways. Taken together, our results suggest that naringin might be an effective agent to treat human cervical cancer.

Al2O3 nanoparticles induce mitochondria-mediated cell death and upregulate the expression of signaling genes in human mesenchymal stem cells.

An increase in the broad usage of Al2O3 nanoparticles (ANPs) in the food and agricultural sectors may produce rare hazards for human health. The objective of this study was to assess the acute toxicity of ANPs in human mesenchymal stem cells (hMSCs) in vitro. Cell viability, cellular uptake, morphology, and gene expression using quantitative real-time polymerase chain reaction (qRT-PCR) were analyzed. The results indicate that ANPs have a significant and dose-dependent effect on cytotoxicity. Control cells showed a characteristic, homogeneous nuclear staining pattern, whereas ANP-exposed cells showed abnormal nuclear morphological changes such as condensation or fragmentation. An early characteristic of apoptosis was observed in ANP-treated cells. Further confirmation of cell death in hMSCs was observed through increased expression of chosen signaling genes and also decreased expression of Bcl-2 during mitochondria-mediated cell death. Although they provide great advantages in food and agricultural products, the chronic and acute toxicity of ANPs still needs to be assessed carefully.

Regulatory effect of epigallocatechin gallate on the expression of C-reactive protein and other inflammatory markers in an experimental model of atherosclerosis.

The present study sought to evaluate the regulatory effect of epigallocatechin gallate (EGCG) on C-reactive protein (CRP) and other inflammatory markers in rats fed an atherogenic diet. Male albino Wistar rats were divided into three groups of nine each. Group I (normal) rats on normal diet received an intraperitoneal (i.p.) injection of physiological saline; group II (atherosclerotic-untreated) rats received an atherogenic diet for 45 days and daily i.p. administration of saline from days 31 to 45; group III (atherosclerotic-treated) rats received an atherogenic diet for 45 days and daily i.p. administration of EGCG (100mg/kg BW) from days 31 to 45 and were then euthanized. The biochemical parameter, CRP, and haematological parameters of inflammation (erythrocyte sedimentation rate [ESR], total leucocyte count [WBC], differential leucocyte count, platelet count) were determined. Immunoblotting and RT-PCR were employed to elucidate the CRP protein and its mRNA expression. Group II rats showed a significant increase in the mean serum CRP level, ESR and total WBC, platelet and differential leucocyte counts, when compared with corresponding values in group I rats. Significantly lower mean levels/counts of inflammatory markers were noted in group III rats than corresponding values in group II rats. Group II rats exhibited significantly higher mean values of CRP protein expression and relative transcript levels of CRP than group I or group III rats. These results suggest that EGCG, a major component of green tea catechins, may decrease the risk of cardiovascular disease by reducing inflammatory markers in rats fed an atherogenic diet.

Prevention of selenite-induced cataractogenesis by rutin in Wistar rats.

PURPOSE: To investigate whether rutin retards selenite-induced cataractogenesis in Wistar rat pups. METHODS: On postpartum day ten, Group I rat pups received an intraperitoneal injection of saline. Group II and III rat pups received a subcutaneous injection of sodium selenite. Group III also received an intraperitoneal injection of rutin once daily on postpartum days 9-14. Both eyes of each pup were examined from day 16 up to postpartum day 30. After sacrifice, extricated pup lenses were analyzed for mean activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase, and glutathione reductase. In addition, the mean concentrations of reduced glutathione (GSH) and of malondialdehyde were analyzed in samples of lenses and hemolysate. RESULTS: There was dense lenticular opacification in all of Group II, minimal opacification in 33.3% of Group III, no opacification in 66.7% of Group III, and no opacification in Group I. Significantly lower mean activities of lenticular antioxidant enzymes were noted in Group II, compared to Group I and III. Significantly lower mean concentrations of GSH and higher mean concentrations of malondialdehyde were noted in samples of hemolysate and lens from Group II, compared to the values in Group I and III. CONCLUSION: Rutin prevents experimental selenite-induced cataractogenesis in rat pups, possibly by preventing depletion of antioxidant enzymes and of GSH, and by inhibiting lipid peroxidation.

Green tea catechins, alleviate hepatic lipidemic-oxidative injury in Wistar rats fed an atherogenic diet.

In the present study, the efficacy of green tea catechins (GTC from the plant Camellia sinensis), with epigallocatechin gallate (EGCG), as the major component, was studied in relation to hepatic oxidative abnormalities in atherosclerotic rats. When male albino Wistar rats were fed an atherogenic diet for 30 days and then treated with saline for 7 or 15 days, there was a significant decline in hepatic mean activities of antioxidant enzymes (catalase, superoxide dismutase, glutathione peroxidase and glutathione-S-transferase), and non-enzymatic antioxidants (reduced glutathione, vitamins C and E) while there was a significant elevation in the mean level of hepatic malondialdehyde (MDA), in comparison to the values noted in control rats fed a normal diet. In addition, a concomitant increase in the activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) was noted, when compared to the values in control rats. Following intraperitoneal administration of GTC (100mg/kg) for 7 or 15 days to rats fed the atherogenic diet, significantly higher mean activities of enzymatic and non-enzymatic antioxidants and lower mean levels of MDA in hepatic tissue and lower mean activities of AST, ALT, ALP and LDH in serum were observed, compared to the values in the rats fed the atherogenic diet and treated with saline. Histopathological studies were performed to provide direct evidence of the atherogenic diet-induced hepatic changes and of the hepatoprotective effect of GTC. These results suggest that EGCG as a major component of green tea catechins may protect against the hepatic abnormalities occurring in Wistar rats fed an atherogenic diet.

Epigallocatechin gallate improves serum lipid profile and erythrocyte and cardiac tissue antioxidant parameters in Wistar rats fed an atherogenic diet.

Oxidative stress is believed to contribute to the pathogenesis of hypercholesterolaemic atherosclerosis; hence, various antioxidant compounds are being evaluated for potential anti-atherogenic effects. The present study assessed the efficacy of epigallocatechin gallate (EGCG), an antioxidant component of the plant Camellia sinensis, in improving serum lipid profile and antioxidant parameters in erythrocytes and cardiac tissue in rats fed an atherogenic diet. In male albino Wistar rats fed an atherogenic diet for 30 days, significantly increased serum levels of total cholesterol, triglycerides and lipoprotein cholesterol fractions and cardiac risk ratio were noted, compared with levels in rats fed a normal diet. Intraperitoneal administration of EGCG (100 mg/kg) for 7 or 15 days to the atherogenic diet-fed rats resulted in significantly lower serum levels of total cholesterol, triglycerides, low-density and very low density lipoprotein cholesterol fractions and a significantly higher serum level of high-density lipoprotein cholesterol compared with levels in atherogenic diet-fed, saline-treated rats. Significantly higher mean malondialdehyde levels and significantly lower mean activities of antioxidant enzymes and mean levels of non-enzymatic antioxidants occurred in atherogenic diet-fed rats compared with those fed a normal diet. When atherogenic diet-fed rats received EGCG treatment for 7 or 15 days, significantly lower mean levels of MDA, higher mean levels of non-enzymatic antioxidants and higher mean activities of enzymatic antioxidants occurred, compared with those in saline-treated rats. Thus, EGCG appears to ameliorate disruptions of serum lipid profile and of antioxidant parameters in erythrocyte and cardiac tissue of Wistar rats fed an atherogenic diet; these results may be relevant to treating human atherosclerosis.

Prevention of selenite-induced cataractogenesis in Wistar rats by the polyphenol, ellagic acid.

The present study sought to evaluate the efficacy of the naturally-occurring polyphenol, ellagic acid, in preventing selenite-induced cataractogenesis. In the present study, Wistar rat pups were divided into 3 groups of 15 each. Group I (normal) rats received an intraperitoneal (i.p.) injection of normal saline on postpartum day 10; group II (cataract-untreated) rats received a single subcutaneous (s.c.) injection of sodium selenite (19 micromol/kg body weight) on postpartum day 10; group III (cataract-treated) pups received a single s.c. injection of sodium selenite on postpartum day 10 and intraperitoneal injections of ellagic acid (200mg/kg body weight) on postpartum days 9-14. At the end of the study period (30th postpartum day), slit-lamp examination of both eyes of each rat pup revealed no lenticular opacification (cataract stage 0) in all eyes of group I pups, definite nuclear cataracts (stages 4-6) in the eyes of all (100%) group II rat pups and no lenticular opacification in eight (53%) and mild lenticular opacification (cataract stages 1-3) in seven (47%) of group III rats (changes in group II vs group III, P<0.01). The mean activities of the antioxidant enzymes catalase, glutathione peroxidase, superoxide dismutase and glutathione-S-transferase were significantly lower in lenses of Group II rats than in Group I or Group III rat lenses. In addition, the mean levels of GSH in lenses and erythrocytes were also significantly lower in Group II rats than in Group I or Group III rats. Conversely, the mean concentration of MDA (an indicator of lipid peroxidation) in lenses and erythrocytes was found to be significantly higher in Group II rats than that in Group I or Group III rats. Also, the mean concentration of calcium was found to be significantly higher in lenses of Group II rats than in those of Group I and Group III rats. The results suggest that ellagic acid can prevent or retard experimental selenite-induced cataractogenesis in Wistar rats. This protective effect in rat lenses appears to occur by maintaining the antioxidant defense system and inhibition of lipid peroxidation.

(3aS,9bR)-Methyl 1-methyl-3-phenyl-1,2,3,3a,4,9b-hexa-hydro-chromeno[4,3-b]pyrrole-3a-carboxyl-ate.

In the title compound, C(20)H(21)NO(3), the heterocyclic six-membered ring adopts a half-chair conformation and the pyrrolidine ring adopts an envelope conformation. The mol-ecular conformation is stabilized by C-H⋯O and C-H⋯N inter-actions.

Acetyl-L-carnitine prevents selenite-induced cataractogenesis in an experimental animal model.

PURPOSE: To investigate whether acetyl-L-carnitine (ALCAR) retards selenite-induced cataractogenesis in vivo. METHODS: On postpartum day 10, group I pups received intraperitoneal saline and group II and group III pups received subcutaneous sodium selenite; Group III pups also received intraperitoneal ALCAR once daily on postpartum days 9-14. Both eyes of each pup were examined up to postpartum day 30. After sacrifice, extricated pup lenses were analyzed for antioxidant and redox system components. RESULTS: There was dense lenticular opacification in all group II pups, minimal opacification in 33% of group III pups, and no opacification in 67% of group III and in all group I pups. Group II lenses exhibited significantly lower values of antioxidant and redox system components and higher malondialdehyde concentrations than group I or group III lenses. CONCLUSION: ALCAR prevents selenite-induced cataractogenesis in Wistar rat pups, possibly by inhibiting depletion of antioxidant enzyme and redox system components and inhibiting lipid peroxidation.

cis-3-Methyl-1-phenyl-8a,9,10,11,12,12a,12b-hexa-hydro-1H,3bH-pyrazolo[3,4:2',3']pyrano[4',5',6'-kl]xanthene.

The asymmetric unit of the title compound, C(23)H(22)N(2)O(2), contains two independent mol-ecules, A and B. The cyclo-hexane ring of mol-ecule B is disordered, with occupancies for the major and minor conformers of 0.570 (9) and 0.430 (9), respectively. The cyclo-hexane ring adopts a boat conformation in mol-ecule A and in the major conformer of mol-ecule B, and a chair conformation in the minor conformer of mol-ecule B. In both independent mol-ecules, one of the dihydro-pyran rings adopts a boat conformation while the other is in a half-chair conformation. The dihedral angle between the pyrazole and phenyl rings is 16.0 (1)° in mol-ecule A and 12.9 (1)° in mol-ecule B. The crystal packing is stabilized by C-H⋯O and C-H⋯N inter-molecular hydrogen bonds.

(3aRS,9bSR)-3-(4-Chloro-phen-yl)-1-methyl-1,2,3,3a,4,9b-hexa-hydro-chromeno[4,3-b]pyrrole-3a-carbonitrile.

In the mol-ecule of the title compound, C(19)H(17)ClN(2)O, the heterocyclic six-membered ring adopts a half-chair conformation, while the pyrrolidine ring has an envelope conformation. In the crystal structure, C-Cl⋯π [Cl⋯centroid = 3.680 (2) Å] inter-actions and van der Waals forces are present.

Prevention of selenite-induced cataractogenesis by acetyl-L-carnitine: an experimental study.

Several studies have suggested that antioxidants retard the process of cataractogenesis by scavenging free oxygen radicals. The present study sought to assess the efficacy of the antioxidant acetyl-L-carnitine (ALCAR) in preventing selenite-induced cataractogenesis in an experimental setting. The first, in vitro phase of the study was performed on lenses from Wistar rats incubated for 24 h at 37 degrees C in Dulbecco's Modified Eagle Medium (DMEM) alone (control, Group I), or in DMEM containing 100 microM of selenite (Group II) or in DMEM containing 100 microM of selenite and 200 microM/ml ALCAR added at the same time as selenite (Group IIIa) or 30 min, 1 h or 2 h later (Groups IIIb, IIIc and IIId, respectively). Gross morphological examination of these lenses revealed dense opacification (cataract formation) in Group II, minimal opacification in some Group IIIa lenses and no opacification in Group I. The mean activities of the antioxidant enzymes catalase and glutathione peroxidase were significantly lower in Group II than in Group I or Group IIIa lenses, while malondialdehyde concentration (an indicator of lipid peroxidation) was significantly higher in Group II lenses than that in Group I or Group IIIa lenses. The second, in vivo phase of the study revealed dense opacification (cataract formation) in 100% of Wistar rat pups receiving subcutaneous sodium selenite alone (19 microM/kg body weight) but in only 37.5% of those receiving subcutaneous selenite and intraperitoneal ALCAR. These data suggest that ALCAR is able to significantly retard experimental selenite-induced cataractogenesis.

Antioxidant activity of the oyster mushroom, Pleurotus ostreatus, on CCl(4)-induced liver injury in rats.

This study was undertaken to investigate the putative antioxidant activity of the oyster mushroom Pleurotus ostreatus on CCl(4)-induced liver damage in male Wistar rats. Intraperitoneal administration of CCl(4) (2ml/kg) to rats for 4 days resulted in significantly elevated (p<0.05) serum levels of glutamic oxaloacetic transaminase (SGOT), glutamic pyruvate transaminase (SGPT) and alkaline phosphatase (SALP) compared to controls. In the liver, significantly elevated levels (p<0.05) of malondialdehyde (MDA) and lowered levels (p<0.05) of reduced glutathione (GSH) were observed following CCl(4) administration. Quantitative and qualitative analysis of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (Gpx) revealed lower activities of these antioxidant enzymes in the liver of CCl(4)-administered rats. An analysis of the isozyme pattern of these enzymes revealed variations in relative concentration presumably due to hepatotoxicity. When rats with CCl(4)-induced hepatotoxicity were treated with the extract of P. ostreatus, the serum SGOT, SGPT and SALP levels reverted to near normal, while the hepatic concentration of GSH, CAT, SOD and Gpx were significantly increased (p<0.05) and that of MDA significantly (p<0.05) lowered, when compared to CCl(4)-exposed untreated rats. Histopathological studies confirmed the hepatoprotective effect conferred by the extract of P. ostreatus. These results suggest that an extract of P. ostreatus is able to significantly alleviate the hepatotoxicity induced by CCl(4) in the rat.