The Modulation of Water, Nitrogen, and Phosphorous Supply for Growth Optimization of the Evergreen Shrubs Ammopiptanthus mongolicus for Revegetation Purpose.

Surface mining is a critical anthropogenic activity that significantly alters the ecosystem. Revegetation practices are largely utilized to compensate for these detrimental impacts of surface mining. In this study, we investigated the effects of five water (W) regimes [W40: 40%, W48: 48%, W60: 60%, W72: 72%, and W80: 80% of field capacity (FC)], five nitrogen (N) (N0: 0, N24: 24, N60: 60, N96: 96, and N120: 120 mg kg-1 soil), and five phosphorus (P) fertilizer doses (P0: 0, P36: 36, P90: 90, P144: 144, and P180: 180 mg kg-1 soil) on morpho-physiological and biochemical parameters of Ammopiptanthus mongolicus plants to assess the capability of this species to be used for restoration purposes. The results showed that under low W-N resources, A. mongolicus exhibited poor growth performance (i.e., reduced plant height, stem diameter, and dry biomass) in coal-degraded spoils, indicating that A. mongolicus exhibited successful adaptive mechanisms by reducing its biomass production to survive long in environmental stress conditions. Compared with control, moderate to high W and N-P application rates greatly enhanced the net photosynthesis rates, transpiration rates, water-use efficiency, chlorophyll (Chl) a, Chl b, total Chl, and carotenoid contents. Under low-W content, the N-P fertilization enhanced the contents of proline and soluble sugar, as well as the activities of superoxide dismutase, catalase, and peroxidase in leaf tissues, reducing the oxidative stress. Changes in plant growth and metabolism in W-shortage conditions supplied with N-P fertilization may be an adaptive strategy that is essential for its conservation and restoration in the desert ecosystem. The best growth performance was observed in plants under W supplements corresponding to 70% of FC and N and P doses of 33 and 36 mg kg-1 soil, respectively. Our results provide useful information for revegetation and ecological restoration in coal-degraded and arid-degraded lands in the world using endangered species A. mongolicus.

New n-nonadecanoyl-ß-sitosterol and other constituents from the stem-bark of Anacardium occidentale.

A new steroidal ester bearing n-nonadecanoyl moiety (1) and a mixture of isomeric cerebrosides (2) along with two known compounds were isolated from the methanol extract of the stem-bark of Anacardium occidentale. The structure of the new steroidal ester was determined as 3-n-nonadecanoyl-ß-sitosterol on the basis of modern spectroscopic techniques (IR, ESI-MS, HR-ESIMS, 1D and 2D NMR) and chemical degradation studies. The structures of the known compounds were identified as gallic acid and tanacetene by comparison of the spectroscopic data with those of reported data. The mixture of cerebrosides was confirmed based on the analysis of 1D and 2D NMR. These compounds were evaluated for cytotoxicity against human cancer cell lines A549, SCOV3 and rat normal cell line NRK49f.

Biomarkers of oxidative stress for in vivo assessment of toxicological effects of iron oxide nanoparticles.

Iron oxide nanoparticles (Fe2O3-IONPs) have revolutionized the industry by significant economic and scientific impacts. Enormous increase in the usage of IONPs has raised concerns about their unseen adverse effects. In the current study, we investigated the effects of IONPs and its bulk on oxidative stress biomarkers, histopathology and biodistribution in rats after 28 days repeated oral treatment at 30, 300 and 1000 mg/kg body weight (b.w.). IONPs size in dry, wet forms and crystallinity was determined using TEM, DLS and XRD. The investigation of oxidative stress biomarkers demonstrated significant increase in lipid peroxidation and decrease in reduced glutathione content in the liver, kidney and the brain of the treated groups in a dose dependant manner. Further, antioxidant enzymes catalase, glutathione S transferase, glutathione peroxidase and glutathione reductase activities were significantly elevated along with significant decrease in superoxide dismutase activity in treated rat organs. ICP-OES analysis revealed dose and size dependant accumulation of IONPs in the liver followed by kidney and the brain than bulk. Moreover, accumulation of IONPs at high dose brought pathological changes only in liver. A large fraction of IONPs was eliminated in urine. Bulk material was substantially excreted in faeces than IONPs suggesting increased absorption of IONPs. In conclusion accumulated IONPs and bulk in organs trigger free radical generation, leading to the induction of oxidative stress condition in rats. The results obtained highlight the importance of toxicity assessments in evaluating the efficiency of IONPs for the safe implementation for diversified applications.

In vitro mutagenicity assessment of aluminium oxide nanomaterials using the Salmonella/microsome assay.

The aim of the current study was to evaluate the potential mutagenicity of aluminium oxide nanomaterials (NMs) (Al(2)O(3)-30 nm and Al(2)O(3)-40 nm). Characterization of the NMs was done before the initiation of the study. The mutagenicity of the NMs was studied by the Ames test with Salmonella typhimurium TA100, TA1535, TA98, TA97a and TA102 strains, in the presence and absence of the S9 mixture. Based on a preliminary cytotoxicity study conducted on the strains, different concentrations of Al(2)O(3)-30 nm, Al(2)O(3)-40 nm and Al(2)O(3)-bulk were selected. At all the concentrations tested, Al(2)O(3)-30 nm and Al(2)O(3)-40 nm did not significantly increase the number of revertant colonies compared to the Al(2)O(3)-bulk and control with or without S9 mixture. Our findings suggest that Al(2)O(3) NMs were devoid of any size and concentration dependent mutagenicity compared to the Al(2)O(3)-bulk and control.

Evaluation of genotoxic effects of oral exposure to aluminum oxide nanomaterials in rat bone marrow.

Nanomaterials have novel properties and functions because of their small size. The unique nature of nanomaterials may be associated with potentially toxic effects. The aim of this study was to evaluate the in vivo genotoxicity of rats exposed with Aluminum oxide nanomaterials. Hence in the present study, the genotoxicity of Aluminum oxide nanomaterials (30 and 40 nm) and its bulk material was studied in bone marrow of female Wistar rats using chromosomal aberration and micronucleus assays. The rats were administered orally with the doses of 500, 1000 and 2000 mg/kg bw. Statistically significant genotoxicity was observed with Aluminum oxide 30 and 40 nm with micronucleus as well as chromosomal aberration assays. Significantly (p < 0.05 or p < 0.001) increased frequency of MN was observed with 1000 and 2000 mg/kg bw dose levels of Aluminum oxide 30 nm (9.4 +/- 1.87 and 15.2 +/- 2.3, respectively) and Aluminum oxide 40 nm (8.1 +/- 1.8 and 13.9 +/- 2.21, respectively) over control (2.5 +/- 0.7) at 30 h. Likewise, at 48 h sampling time a significant (p < 0.05 or p < 0.001) increase in frequency of MN was evident at 1000 and 2000 mg/kg bw dose levels of Aluminum oxide 30 nm (10.6 +/- 1.68 and 16.6 +/- 2.66, respectively) and Aluminum oxide 40 nm (9.0 +/- 1.38 and 14.7 +/- 1.68, respectively) compared to control (1.8 +/- 0.75). Significantly increased frequencies (p < 0.05 or p < 0.001) of chromosomal aberrations were observed with Aluminum oxide 30 nm (1000 and 2000 mg/kg bw) and Aluminum oxide 40 nm (2000 mg/kg bw) in comparison to control at 18 and 24 h. Further, since there is need for information on the toxicokinetics of nanomaterials, determination of these properties of the nanomaterials was carried out in different tissues, urine and feces using inductively coupled plasma mass spectrometry (ICP-MS). A significant size dependent accumulation of Aluminum oxide nanomaterials occurred in different tissues, urine and feces of rats as shown by ICP-MS data. The results of our study suggest that exposure to Aluminum oxide nanomaterials has the potential to cause genetic damage.

In vivo assessment of genotoxic effects of Annona squamosa seed extract in rats.

Widespread use of pesticides represents a potential risk to human and environmental health. Hence, biopesticides from plants are some of the future strategies for plant protection. In this regard, a seed extract of Annona squamosa was prepared and found to be a promising pesticide. In order to establish the inherent toxicity and non-target safety required for registration and marketing of pesticides, toxicological studies are conducted. The genotoxicity potential was evaluated in rats with 75, 150 and 300 mg/kg Annona squamosa by the comet assay in leucocytes, micronucleus and chromosomal aberration tests in bone marrow. We also studied the effects of 300 mg/kg of extract on lipid peroxidation, reduced glutathione level and glutathione S transferase activity in liver, lungs, brain, kidneys, heart and spleen of treated rats. The comet assay showed a statistically significant dose related increase in DNA migration. The micronucleus and chromosomal aberration tests revealed a significant induction in frequency of micronuclei and chromosomal aberrations at 150 and 300 mg/kg. Annona squamosa treatment significantly enhanced lipid peroxidation, decreased glutathione and glutathione S transferase levels revealing the oxidative stress condition. Our results warrant careful use of Annona squamosa seed extract as a biopesticide till more tests are carried out.

In vivo genotoxicity assessment of aluminium oxide nanomaterials in rat peripheral blood cells using the comet assay and micronucleus test.

Advances in nanotechnology and its usage in various fields have led to the exposure of humans to engineered nanomaterials (NMs) and there is a need to tackle the potential human health effects before these materials are fully exploited. The main purpose of the current study was to assess whether aluminium oxide NMs (Al(2)O(3)-30 nm and Al(2)O(3)-40 nm) could cause potential genotoxic effects in vivo. Characterization of Al(2)O(3)-30 nm and Al(2)O(3)-40 nm was done with transmission electron microscopy, dynamic light scattering and laser Doppler velocimetry prior to their use in this study. The genotoxicity end points considered in this study were the frequency of micronuclei (MN) and the percentage of tail DNA (% Tail DNA) migration in rat peripheral blood cells using the micronucleus test (MNT) and the comet assay, respectively. Genotoxic effects were evaluated in groups of female Wistar rats (five per group) after single doses of 500, 1000 and 2000 mg/kg body weight (bw) of Al(2)O(3)-30 nm, Al(2)O(3)-40 nm and Al(2)O(3)-bulk. Al(2)O(3)-30 nm and Al(2)O(3)-40 nm showed a statistically significant dose-related increase in % Tail DNA for Al(2)O(3)-30 nm and Al(2)O(3)-40 nm (P < 0.05). However, Al(2)O(3)-bulk did not induce statistically significant changes over control values. The MNT also revealed a statistically significant (P < 0.05) dose-dependent increase in the frequency of MN, whereas Al(2)O(3)-bulk did not show any significant increase in frequency of MN compared to control. Cyclophosphamide (40 mg/kg bw) used as a positive control showed statistically significant (P < 0.001) increase in % Tail DNA and frequency of MN. The biodistribution of Al(2)O(3)-30 nm and Al(2)O(3)-40 nm and Al(2)O(3)-bulk in different rat tissues, urine and feces was also studied 14 days after treatment using inductively coupled plasma mass spectrometry. The data indicated that tissue distribution of Al(2)O(3) was size dependent. Our findings suggest that Al(2)O(3) NMs were able to cause size- and dose-dependent genotoxicity in vivo compared to Al(2)O(3)-bulk and control groups.

Effects of dietary vitamin E, C and soybean oil supplementation on antioxidant enzyme activities in liver and muscles of rats.

The effect of elevated levels of dietary vitamin E, C and a combination of vitamin E and C (E&C) with soybean oil on activities of antioxidant (AOE) enzymes important in the protection against lipid peroxidation was studied in male rats fed with vitamin C (12 mg/g), vitamin E (3.68 mg/g) or E&C (3.68 mg/kg+12 mg/g) supplemented diets for 28 days. Catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) activity in liver, pectoralis major (PM) and sartorius (S) muscles was increased significantly in rats fed with dietary vitamin C, E separately, and vitamin C&E combination, except, superoxide dismutase (SOD), which showed no alterations. These results clearly indicated that vitamin E&C separately and E&C together increased AOE activity in liver, PM and S muscle of rats. However, vitamin E and C combination enhanced AOE activity more significantly and our findings suggest the possible role of vitamin C&E and their combination in reducing the risk of chronic diseases related to oxidative stress.

Lipid peroxidation and antioxidant enzyme activity in different organs of mice exposed to low level of mercury.

The effects of mercuric chloride (Hg) on lipid peroxidation (LPO), glutathione reductase (GR), glutathione peroxidase (GPx), superoxide dismutase (SOD) and glutathione (GSH) levels in different organs of mice (CD-1) were evaluated. Mice were exposed (2 days/week) to 0.0 (control), 0.8 (low) and 8.0 (mid) and 80.0 (high) gHg/kg/day for 2 weeks. The high dose group was excluded from the study due to high mortality. LPO levels in kidney, testis and epididymus at low and mid doses; GR and GPx levels in testis at mid dose; SOD levels in brain and testis at both doses, liver and epididymus at mid dose; GSH levels in testis at both doses were significantly increased compared to their controls. However, the GR levels in kidney at both doses and in epididymus at mid dose; GPx levels in kidney and epididymus and SOD levels in kidney at both the doses; GSH levels in epididymus at mid dose were significantly decreased compared to their control. Body weight gain and food efficiency were significantly reduced (p<0.05) in mid dose. These results indicated that Hg treatment enhanced LPO in all tissues, but showed significant enhancement only in kidney, testis and epididymus suggesting that these organs were more susceptible to Hg toxicity. The increase in antioxidant enzyme levels in testis could be a mechanism protecting the cells against reactive oxygen species.

The effect of subacute administration of a neem pesticide on rat metabolic enzymes.

Acute toxicity of a neem pesticide (Vepacide-Tech) was studied in male Wistar rats by oral (single) intubation for 7 days. Vepacide was found to be moderately toxic to rat based on LD50 value. Subacute toxicity of Vepacide-Tech was also studied in male rats by oral (multiple) intubation of low (80 mg Kg-1 day-1), medium (160 mg Kg-1 day-1) and high dose (320 mg Kg-1 day-1) for 90 days. High dose caused a significant decrease in Cytochrome P-450 (Cyt. P-450) concentration at 45 and 90 days and the medium dose caused same effect at 90th day in liver and lung. Kidney showed similar effect at 90 days by the three doses. Cytochrome b5 (Cyt. b5) concentration was significantly decreased in liver, lung and kidney at 45 and 90 days at medium and high doses. Brain Cyt.b5 concentration was decreased on 90th day at high dose. Cytochrome P-450 reductase (Cyt.P-450 reductase) concentration was decreased significantly in liver and brain at 45 and 90 days, respectively at medium and high doses. The withdrawal study (28 days) has shown significant recovery. These results demonstrate that low levels exposure of Vepacide may have significant effect on the xenobiotic detoxification mechanism of different tissues of rat.