Search results for: smoke toxicity

Authors: S. Madhusudhan, G. V. Vaniprabha

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Worldwide statistics have shown that smoking contributes significantly to mortality, with nicotine, being more addictive. Smoking causes more than 7,00,000 deaths/year in India. Compared to the general population, the prevalence of smoking is found to be much higher among people with psychotic disorders and, more so in schizophrenia. Schizophrenic patients who smoke tend to have higher frequency of heavy smoking, with rates ranging from 60% to as high as 80%. Hence, smokers with psychiatric disorders suffer higher rates of morbidity and mortality secondary to smoking related illnesses.

Keywords: brief intervention, nicotine dependence, schizophrenia

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Authors: Mohammad Y. Alfaifi

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Methidathion (MTD) (Trade name Supracide®) is a non-systemic organophosphorus insecticide used intensively worldwide including Saudi Arabia. However, there is a lack in published studies about it's genotoxicity. In this study we evaluated MTD toxicity in rat bone marrow cells (in vivo) and in lymphocytes (in vitro) using different doses based on LD50. MNNCE (Micronucleated normocromatic erythrocytes) and MNPCE (Micronucleated polychromatic erythrocytes), NDI (Nuclear division index) and NDCI (nuclear division cytotoxicity index), necrotic and apoptotic cells were recorded in rat's bone marrow samples. CA, MI (number of cells undergoing mitosis) necrotic, and apoptotic cells recorded in lymphocytes. Results showed that there was a slight increase in the frequency of micronucleated bone marrow cells. However, no structural chromosomal aberrations were detected in vivo or in vitro. On the other hand, the results showed significant increase in necrotic and apoptotic cells following MTD administration in a dose-dependent manner comparing to positive and negative control groups. In light of these results, MTD can be considered highly cytotoxic and moderate genotoxic, and precaution should be taken when using MTD.

Keywords: methidathion, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations

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Authors: Xing Ping Hu, Yuexun Tian, Jerome Hogsette

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Housefly, Musca domestica L., is a serious urban nuisance and public health/food safety concern. This study evaluated the topical toxicity of 17 essential oil components and 3 plant essential oils against permethrin-resistant adult females and insecticide-susceptible house fly strains. Results show that thymol had the lowest LD₅₀ values against permethrin-resistant strain (43.77 and 41.10 ug per fly) and permethrin-susceptible strain (35.19 and 29.16 ug per fly) at both 24- and 48-hours post treatments; (+)-Pulegone had the lowest LD₉₅ values against the permethrin-resistant strain (0.15 and 0.10 mg per fly) at 24- and 48-hours post treatments, whereas plant thyme oil had the lowest LD₉₅ value of 0.17 mg per fly at post-24h and post-48h against the permethrin-susceptible strain. Additionally, the LD₅₀s was slightly but not significantly negatively correlated with the boiling points of the compounds tested; but showed no correlation with the density and LogP. These results indicate that specific essential oils and compounds have topical insecticidal properties against house flies with low dose. They may have the potential for development as botanical insecticides.

Keywords: urban pest, public health, pest management, botanical chemical

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Authors: Afaf Bouchoucha, Karima Si Larbi, Mohamed Amine Bourouaia, Salah.Boulanouar, Safia.Djabbar

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The synthesis, characterization and comparative biological study of manganese (II) and copper (II) complexes with an heterocyclic ligand used in pharmaceutical field (Scheme 1), were reported. Two kinds of complexes were obtained with derivative sulfonamide, [M (L)₂ (H₂O)₂].H₂O and [M (L)₂ (Cl)₂]3H₂O. These complexes have been prepared and characterized by elemental analysis, FAB mass, ESR magnetic measurements, FTIR, UV-Visible spectra and conductivity. Their stability constants have been determined by potentiometric methods in a water-ethanol (90:10 v/v) mixture at a 0.2 mol l-1 ionic strength (NaCl) and at 25.0 ± 0.1 ºC using Sirko program. DFT calculations were done using B3LYP/6-31G(d) and B3LYP/LanL2DZ. The antimicrobial activity of ligand and complexes against the species Escherichia coli, P. aeruginosa, Klebsiella pneumoniae, S. aureus, Bacillus subtilisan, Candida albicans, Candida tropicalis, Saccharomyces, Aspergillus fumigatus and Aspergillus terreus has been carried out and compared using agar-diffusion method. Also, the toxicity study was evaluated on synchesis complexes using Mice of NMRI strain.

Keywords: hetterocyclic ligand, complex, stability constant, antimicrobial activity, DFT, acute and genotoxicity study

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Authors: Naser Kazemi Eilaki, Carolyn Ahmer, Ilona Heldal, Bjarne Christian Hagen

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Older people and people with disabilities are recognized as at-risk groups when it comes to egress and travel from hazard zone to safe places. One's disability can negatively influence her or his escape time, and this becomes even more important when people from this target group live alone. This research deals with the fire safety of mentioned people's buildings by means of probabilistic methods. For this purpose, fire safety is addressed by modeling the egress of our target group from a hazardous zone to a safe zone. A common type of detached house with a prevalent plan has been chosen for safety analysis, and a limit state function has been developed according to the time-line evacuation model, which is based on a two-zone and smoke development model. An analytical computer model (B-Risk) is used to consider smoke development. Since most of the involved parameters in the fire development model pose uncertainty, an appropriate probability distribution function has been considered for each one of the variables with indeterministic nature. To achieve safety and reliability for the at-risk groups, the fire safety index method has been chosen to define the probability of failure (causalities) and safety index (beta index). An improved harmony search meta-heuristic optimization algorithm has been used to define the beta index. Sensitivity analysis has been done to define the most important and effective parameters for the fire safety of the at-risk group. Results showed an area of openings and intervals to egress exits are more important in buildings, and the safety of people would improve with increasing dimensions of occupant space (building). Fire growth is more critical compared to other parameters in the home without a detector and fire distinguishing system, but in a home equipped with these facilities, it is less important. Type of disabilities has a great effect on the safety level of people who live in the same home layout, and people with visual impairment encounter more risk of capturing compared to visual and movement disabilities.

Keywords: fire safety, at-risk groups, zone model, egress time, uncertainty

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Authors: Sameena Malik, Ghosh Prakash, Sandeep Mudliar, Vishal Waindeskar, Atul Vaidya

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In this study, the biodegradability enhancement along with COD color and toxicity removal of pharmaceutical effluent by O₃, O₃/Fe²⁺, O₃/nZVI processes has been evaluated. The nZVI particles were synthesized and characterized by XRD and SEM analysis. Kinetic model was reasonably developed to select the ozone doses to be applied based on the ozonation kinetic and mass transfer coefficient values. Nano catalytic ozonation process (O₃/nZVI) effectively enhanced the biodegradability (BI=BOD₅/COD) of pharmaceutical effluent up to 0.63 from 0.18 of control with a COD, color and toxicity removal of 62.3%, 93%, and 75% respectively compared to O₃, O₃/Fe²⁺ pretreatment processes. From the GC-MS analysis, 8 foremost organic compounds were predominantly detected in the pharmaceutical effluent. The disappearance of the corresponding GC-MS spectral peaks during catalyzed ozonation process indicated the degradation of the effluent. The changes in the FTIR spectra confirms the transformation/destruction of the organic compounds present in the effluent to new compounds. Subsequent aerobic biodegradation of pretreated effluent resulted in biodegradation rate enhancement by 5.31, 2.97, and 1.22 times for O₃, O₃/Fe²⁺ and O₃/nZVI processes respectively.

Keywords: iron nanoparticles, pharmaceutical effluent, ozonation, kinetics, mass transfer

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Authors: Muneesh Kumar, Rajesh Kumar, Sangeeta Devi

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Heavy metals are a major problem because they are toxic and tend to accumulate in living organisms. This study was carried out with the aims of studying on histopathology of Zn(SO4) toxicity on gill and liver tissues of catfish (Clarias batrachus) within the period of 96 h. Totally, 140 fishes with mean weight 50±10 g were stocked in 12 aquariums with capacity of 200 L water and divided in to 3 trails including control, 4 ppm and 8 ppm of Zn with 3 replicates. Tissue samples were fixed by bouin’s solution and sectioned in 7 μm based on histological regular method and stained with Hematoxylin and Eosin (H&E) method for microscopic study within the period of 96 h. Results showed some damaged such as hyperplasia, telangiectasis and edema, necrosis of second filaments, jerky movement, aneurism, hyperemia and fusion of second filaments in gills; and cell atrophy, necrosis, fatty degeneration, hyperemia and bile stagnation at different treatments in comparison with control. Gill and liver tissue damages were severed with the increase of Zn concentration and days. Therefore, Zn had acute toxicity effects on gill and liver tissues in Catfish at 5 and 10 ppm concentrations.

Keywords: gill, liver, histopathology, zinc, Clarias batrachus

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Authors: Justina I. R. Udotong, Ofonime U. M. John

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Toxicity of copper (Cu), lead (Pb) and iron (Fe) to Tilapia guinensis was carried out for 4 days with a view to determining their effects on the liver and muscle tissues. Tilapia guinensis samples of about 10 - 14cm length and 0.2 – 0.4kg weight each were obtained from University of Calabar fish ponds and acclimated for three (3) days before the experimental set up. Survivors after the 96-hr LC50 test period were selected from test solutions of the heavy metals for the histopathological studies. Histological preparations of liver and muscle tissues were randomly examined for histopathological lesions. Results of the histological examinations showed gross abnormalities in the liver tissues due to pathological and degenerative changes compared to liver and muscle tissues from control samples (tilapia fishes from aquaria without heavy metals). Extensive hepatocyte necrosis with chronic inflammatory changes was observed in the liver of fishes exposed to Cu solution. Similar but less damaging effects were observed in the liver of fishes exposed to Pb and Fe. The extent of lesion observed was therefore heavy metal-related. However, no pathologic changes occurred in the muscle tissues.

Keywords: degenerative changes, heavy metal, hepatocyte necrosis, histopathology, toxicity

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Authors: Kuladip Jana, Bhaswati Banerjee, Parimal C. Sen

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Benzo(a)pyrene [B(a)P] is an environmental toxicant present mostly in cigarette smoke and car exhaust, is an aryl hydrocarbon receptor (AhR) ligand that exerts its toxic effects on both male and female reproductive systems along with carcinogenesis in skin, prostate, ovary, lung and mammary glands. Our study was focused on elucidating the molecular mechanism of B(a)P induced male reproductive toxicity and its prevention with phytochemical like resveratrol. In this study, the effect of B(a)P at different doses (0.1, 0.25, 0.5, 1 and 5 mg /kg body weight) was studied on male reproductive system of Wistar rat. A significant decrease in cauda epididymal sperm count and motility along with the presence of sperm head abnormalities and altered epididymal and testicular histology were documented following B(a)P treatment. B(a)P treatment resulted apoptotic sperm cells as observed by TUNEL and Annexin V-PI assay with increased Reactive Oxygen Species (ROS), altered sperm mitochondrial membrane potential (ΔΨm) with a simultaneous decrease in the activity of antioxidant enzymes and GSH status. TUNEL positive apoptotic cells also observed in testis as well as isolated germ and Leydig cells following B(a)P exposure. Western Blot analysis revealed the activation of p38 mitogen activated protein kinase (p38MAPK), cytosolic translocation of cytochrome-c, upregulation of Bax and inducible nitric oxide synthase (iNOS) with cleavage of poly ADP ribose polymerase (PARP) and down regulation of BCl2 in testis upon B(a)P treatment. The protein and mRNA levels of testicular key steroidogenesis regulatory proteins like steroidogenic acute regulatory protein (StAR), cytochrome P450 IIA1 (CYPIIA1), 3β hydroxy steroid dehydrogenase (3β HSD), 17β hydroxy steroid dehydrogenase (17β HSD) showed a significant decrease in a dose dependent manner while an increase in the expression of cytochrome P450 1A1 (CYP1A1), Aryl hydrocarbon Receptor (AhR), active caspase- 9 and caspase- 3 following B(a)P exposure. We conclude that exposure of benzo(a)pyrene caused testicular gamatogenic and steroidogenic disorders by induction of oxidative stress, inhibition of StAR and other steroidogenic enzymes along with activation of p38MAPK and initiated caspase-3 mediated germ and Leydig cell apoptosis. Next we investigated the role of resveratrol on B(a)P induced male reproductive toxicity. Our study highlighted that resveratrol co-treatment with B(a)P maintained testicular redox potential, increased serum testosterone level and prevented steroidogenic dysfunction with enhanced expression of major testicular steroidogenic proteins (CYPIIA1, StAR, 3β HSD,17β HSD) relative to treatment with B(a)P only. Resveratrol suppressed B(a)P-induced testicular activation of p38 MAPK, ATF2, iNOS and ROS production; cytosolic translocation of Cytochome c and Caspase 3 activation thereby prevented oxidative stress of testis and inhibited apoptosis. Resveratrol co-treatment also decreased B(a)P-induced AhR protein level, its nuclear translocation and subsequent CYP1A1 promoter activation, thereby decreased protein and mRNA levels of testicular cytochrome P4501A1 (CYP1A1) and prevented BPDE-DNA adduct formation. Our findings cumulatively suggest that resveratrol prevents activation of B(a)P by modulating the transcriptional regulation of CYP1A1 and acting as an antioxidant thus prevents B(a)P-induced oxidative stress and testicular apoptosis.

Keywords: benzo(a)pyrene, resveratrol, testis, apoptosis, cytochrome P450 1A1 (CYP1A1), aryl hydrocarbon receptor (AhR), p38 MAPK/ATF2/iNOS

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Authors: Monthana Weerawatanakorn, Hajime Tamaki, Yonathan Asikin, Koji Wada, Makoto Takahashi, Chi-Tang Ho, Min-Hsiung Pan

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Rice bran and sugarcane are significant sources of wax containing policosanol (PC), the cholesterol-lowering nutraceutical available in the market. The processing of rice bran oil causes the loss of PC content into various waste products. Therefore, we hypothesise that defatted rice bran (DRB) as agricultural waste product and rice bran oil (RBO) retain a varying but significant amount of PC wax. Non-centrifugal cane sugar (NCS) or cane brown sugar has been consumed worldwide and possesses various health benefits. Since PC wax is mainly in the outer layer rinds of cane, PC contents of the granulated sugar are reduced due to the peeling step. The study aimed to increase PC contents of the granular brown sugar by adding wax extracted from DRB and RBO and to investigate the toxicity of the developed products. The results showed that the total PC contents including long chain aldehyde of products were increased to the maximum level of 147.97 mg/100 g and 40.14 mg/100 g for extracted wax and rice bran oil addition, respectively. PC content of RBO was found to be 96.93 mg/100 g. DRB is promising source of policosanol (6,044.7 mg/100 g). The 28-day toxicity evaluations of the developed sugar revealed no adverse effects on the liver, spleen or kidney.

Keywords: enrichment, sugarcane, policosanol, defatted rice bran, wax

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Authors: Sonam Agarwal, Renu Bist

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Current study emphasizes the toxic effects of dichlorvos on serum in terms of oxidative stress. Meanwhile, a protective action of bacoside A and bromelain was investigated against the biochemical alterations in serum. The experimental design included six groups of mice: saline was given as a vehicle to the control mice (group I). Mice belonging to groups II, III and IV, were administered with dichlorvos (40 mg/kg b.w.), bromelain and bacoside A, respectively. The fifth group received a combination of bromelain and bacoside A. In group VI, Bacoside A, and bromelain were administered 20 minutes prior to exposure of dichlorvos. Thiobarbituric acid reactive substances (TBARS), protein carbonyl content (PCC), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and reduced glutathione (GSH) level were used as biochemical test of toxic action for dichlorvos intoxication. Significantly increased TBARS and PCC level in second group suggests that dichlorvos enhances the production of free radicals in serum of mice (p< 0.05). Antioxidants treatment significantly decreased the levels of TBARS and PCC (p< 0.05). Dichlorvos administration causes a significant reduction in the level of CAT, SOD, GPx and GSH (p< 0.05) which was restored significantly by co-administration of bromelain and Bacoside A in dichlorvos exposed mice (p< 0.05). Treatment of bromelain and Bacoside A in combination served as better scavengers of toxicity induced by dichlorvos.

Keywords: bacoside A, bromelain, dichlorvos, serum

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Authors: C. E. Ihejirika, M. I. Nwachukwu, R. F. Njoku-Tony, O. C. Ihejirika, U. O. Enwereuzoh, E. O. Imo, D. C. Ashiegbu

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Disposal of industrial solid wastes in the environment is a major environmental challenge. This study investigated the effects of calcium carbide waste dumpsites on soil quality. Soil samples were collected with hand auger from three different dumpsites at varying depths and made into composite samples. Samples were subjected to standard analytical procedures. pH varied from 10.38 to 8.28, nitrate from 5.6mg/kg to 9.3mg/kg, phosphate from 8.8mg/kg to 12.3mg/kg, calcium carbide reduced from 10% to to 3%. Calcium carbide was absent in control soil samples. Bacterial counts from dumpsites ranged from 1.8 x 105cfu/g - 2.5 x 105cfu/g while fungal ranged from 0.8 x 103cfu/g - 1.4 x 103cfu/g. Bacterial isolates included Pseudomonas spp, Flavobacterium spp, and Achromobacter spp, while fungal isolates include Penicillium notatum, Aspergillus niger, and Rhizopus stolonifer. No organism was isolated from the dumpsites at soil depth of 0-15 cm, while there were isolates from other soil depths. Toxicity might be due to alkaline condition of the dumpsite. Calcium carbide might be bactericidal and fungicidal leading to cellular physiology, growth retardation, death, general loss of biodiversity and reduction of ecosystem processes. Detoxification of calcium carbide waste before disposal on soil might be the best option in management.

Keywords: biodiversity, calcium-carbide, denitrification, toxicity

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Authors: A. B. Amna, M. A. E. Samia, A. K. Hassan

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The clinical, pathological, hematological and biological changes in Newzealand rabbits groups given daily oral doses of 0.1,0.25 and 0.5 ml/kg body weight/day of Cpmbopogon proximus oil extract were investigated in an experiment durated for 21 days. Other than the dose co-related mortality rates, the clinical signs were observed daily after dosing to be low appetite and nervous signs including restlessness and increased consciousness. Pulmonary excretion of the oil extract led to bloody spots on the lungs, lymphocyte infiltration, congestion and edema. Renal glumeruli manifested lymphocyte infiltration in addition to shrinkages and easinophilic material in the medulla, if considered with the corticomedullary generalized necrosis and the significant changes in urea, they can explain the renal dysfunction. Hepatic malfunction was manifested by significant changes in serum alkaline phosphatase and aspartate transferases accompanied by the congested, fatty changed livers. The direct physical effect of the extracted oil was detected by the catarrhal inflammation of the intestines.There was no significant haematological change except for the slight changes in RBCs and MCVs in rabbits given the highest dose. Future work for Cpmbopogon proximus oil extract was forwarded and practical implications of the result were highlighted.

Keywords: toxicity, cymbopogon proximus (maharaib), oil extract, Newzealand rabbits

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Authors: R. Stevens, W.L. Bryden

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The objective of this study was to determine the efficacy of zeolite in preventing the adverse effects of aflatoxin B1 (AFB1) in broilers. A total of 540 one-day-old Ross chicks were randomly divided into nine treatments, with four replicate pens per treatment and 15 chicks per pen. The treatments included 3 Levels of AFB1 (0,1and 2 mg/kg diet) and 3 levels of zeolite (0, 1.5 and 3 %) in a 3 ×3 factorial arrangement. The experimental treatments commenced on d 7 post-hatch. A starter diet was provided from d 1 to 14, a grower diet from d 15 to 28 and a finisher diet from d 29 to d 49. Diets were based on corn and soybeans and formulated to meet the bird's requirements. The evaluated parameters were as follows: Bodyweight, daily gain, feed intake (FI), feed conversion (FC), relative weights of organs (carcass, liver, heart and abdominal fat) and clinical biochemistry parameters: alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Bodyweight, daily gain and FC were significantly (P<0.05) impaired by aflatoxin. Relative weights of the liver and heart were also affected. The addition of zeolite (1.5 and 3 %) to the contaminated diets ameliorated the effects of aflatoxin, especially at the higher level of inclusion. These data demonstrate that this specific sorbent (zeolite) can protect against the toxicity of AFB1in young broiler chicks.

Keywords: aflatoxin, broiler, toxicity, zeolite

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Authors: Azza A. Ali, Toqa M. Elnahhas, Abeer I. Abd El-Fattah, Mona M. Kamal, Karema Abu-Elfotuh

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Background: Environmental pollution with the different aluminium (Al) containing compounds especially those in industrial waste water exposes people to higher than normal levels of Al that represents an environmental risk factor. Cosmetics, Al ware, and containers are also sources of Al besides some foods and food additives. In addition to its known neurotoxicity, Al affects other body structures like skeletal system, blood cells, liver and kidney. Accumulation of Al in kidney and liver induces nephrotoxicity and hepatotoxicity. Coenzyme Q10 (CoQ10) is a pseudo-vitamin substance primarily present in the mitochondria. It is a powerful antioxidant and acts as radical scavenger. Wheat grass is a natural product that contains carbohydrates, proteins, vitamins, minerals, enzymes and has antioxidant, anti-inflammatory, anticancer and cardiovascular protection activities. Cocoa is an excellent source of iron, potent antioxidants and can protect against many diseases. Vinpocetine is an antioxidant and anti inflammatory while zinc is an essential trace element involved in cell division and its deficiency is observed in many types of liver disease. Objective: To evaluate and compare the potency of different drugs (CoQ10, wheatgrass, cocoa, vinpocetine and zinc) against nephro- and hepato-toxicity induced by Al in rats. Methods: Rats were divided to seven groups and received daily for three weeks either saline for control group or AlCl3 (70 mg/kg, IP) for Al-toxicity model groups. Five groups of Al-toxicity model (treated groups) were orally received together with Al each of the following; CoQ10 (200mg/kg), wheat grass (100mg/kg), cocoa powder (24mg/kg), vinpocetine (20mg/kg) or zinc (32mg/kg). Biochemical changes in the serum level of Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate deshydrogenase (LDH) as well as total bilirubin, lipids, cholesterol, triglycerides, glucose, proteins, creatinine and urea were measured. Liver and kidney specimens from all groups were also collected for the assessment of hepatic and nephrotic level of inflammatory mediators (TNF-α, IL-6β, nuclear factor kappa B (NF-κB), Caspase-3, oxidative parameters (MDA, SOD, TAC, NO) and DNA fragmentation. Histopathological changes in liver and kidney were also evaluated. Results: Three weeks of AlCl3 (70 mg/kg, IP) exposure induced nephro- and hepato-toxicity in rats. Treatment by the all used drugs showed protection against hazards of AlCl3. The protective effects were indicated by the significant decrease in ALT, AST, ALP, LDH as well as total bilirubin, lipids, cholesterol, triglycerides, glucose, creatinine and urea levels which were increased by Al. Liver and kidney of the treated groups showed decrease in MDA, NO, TNF-α, IL-6β, NF-κB, caspase-3 and DNA fragmentation which were increased by Al, together with significant increase in total proteins, SOD and TAC which were decreased by Al. The protection against both nephro- and hepato-toxicity was more pronounced especially with CoQ10 and wheat grass than the other used drugs. Histopathological examinations confirmed the biochemical results of toxicity and of protection. Conclusion: Protection from nephrotoxicity, hepatotoxicity and the consequent degenerations induced by Al can be achieved by using different drugs as CoQ10, wheatgrass, cocoa, vinpocetine and zinc, but CoQ10 as well as wheat grass possesses the most superior protection.

Keywords: aluminum, nephrotoxicity, hepatotoxicity, coenzyme Q10, wheatgrass, cocoa, vinpocetine, zinc

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Authors: Bendjazia Radia, Samira Kilani-Morakchi, Nadia Aribi

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Azadirachtin is a biorational insecticide commonly reported as selective to a range of beneficial insects. It is one of the most biologically active natural inhibitors of insect growth and development and it is known to be an antagonist of the juvenile hormone and 20-hydroxyecdysone (20E). However, its mechanism of action remains still unknown. In the present study, the toxicity of a commercial formulation of Azadirachtin (Neem Azal, 1% azadirachtine) was evaluated by topical application at various doses (0.1, 0.25, 0.5, 1 and 2 µg/insect) on the third instars larvae of D. melanogaster. Lethal doses (LD25: 0.28µg and LD50: 0.67µg), were evaluated by cumulated mortality at the immature stages. The effects of azadirachtin (LD25 and LD50) were then evaluated on the development (duration of the larval and pupal instars, the weight of larvae, pupa and adults) of Drosophila melanogaster. Results showed that the insecticide increased significantly the larval and pupal instar duration. A reduction of larval and pupal weight is noted under azadirachtin treatment as compared to controls. In addition, the weight of surviving adults at the two tested dose was also reduced. In conclusion, azadirachtin seemed to interfere with the functions of the endocrine system resulting in development defects.

Keywords: azadirachtin, d.melanogaster, toxicity, development

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Authors: Mala Tankam Carine, Kekeunou Sévilor, Nukenine Elias

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Storage and preservation of agricultural products remain the only conditions ensuring the almost permanent availability of foodstuffs. However, infestations due to insects and microorganisms often occur. Callosobruchus maculatus is a pest that causes a lot of damage to cowpea stocks in the tropics. Several methods are adopted to limit their damage, but the use of synthetic chemical insecticides is the most widespread. Biopesticides in sustainable agriculture respond to several environmental, economic and social concerns while offering innovative opportunities that are ecologically and economically viable for producers, workers, consumers and ecosystems. Our main objective is to evaluate the insecticidal efficacy of binary combinations of Fossilshield with root-bark powder of Securidaca longepedunculata against Callosobruchus maculatus in stored cowpea Vigna unguiculata. Laboratory bioassays were conducted in stored grains to evaluate the toxicity of root-bark powder of Securidaca longepedunculata alone or combined with diatomaceous earth Fossil-Shield ® against C. maculatus. Twenty-hour-old adults of C. maculatus were exposed to 50g of cowpea seeds treated with four doses (10, 20, 30, and 40g/kg) of root-bark powder of S. longepedunculata, on the one hand, and (0.5, 1, 1.5, and 2 g/kg) on DE and binary combinations on the other hand. 0g/kg corresponded to untreated control. Adult mortality was recorded up to 7 days (d) post-treatment, whereas the number of F1 progeny was assessed after 30 d. Weight loss and germinative ability were conducted after 120 d. All treatments were arranged according to a completely randomized block with four replicates. The combined mixture of S. longepedunculata and DE controlled the beetle faster compared to the root-bark powder of S. longepedunculata alone. According to the Co-toxicity coefficient, additive effect of binary combinations was recorded at 3-day post-exposure time with the mixture 25% FossilShield + 75% S. longepedunculata. A synergistic action was observed after 3-d post-exposure at mixture 50% FossilShield + 50% S. longepedunculata and at 1-d and 3-d post-exposure periods at mixture 75% FossilShield + 25% S. longepedunculata. The mixture 25% FossilShield + 75% S. longepedunculata induced a decreased progeny of 6 times fewer individuals for 4.5 times less weight loss and 2, 9 times more sprouted grains than with root-bark powder of S. longepedunculata. The combination of FossilShield + S. longepedunculata was more potent than root-bark powder of S. longepedunculata alone, although the root-bark powder of S. longepedunculata caused significant reduction of F1 adults compared to the control. Combined action of botanical insecticides with FossilShield as a grain protectant in an integrated pest management approach is discussed.

Keywords: diatomaceous earth, cowpea, callosobruchus maculatus, securidaca longepedunculata, combined action, co-toxicity coefficient

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Authors: Yogesh Bacchaw, Ashish Dabade

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Nowadays oral health is raising concern in society. Acid producing microorganisms changes the oral pH and creates a favorable environment for microbial growth. This growth not only promotes dental decay but also bad breath. Generally Recognized As Safe (GRAS) listed component was incorporated in chewing gum as an antimicrobial agent. The chewing gum produced exhibited up to 9 hours of antimicrobial activity against oral microflora. The toxicity of GRAS component per RACC value of chewing gum was negligible as compared to actual toxicity level of GRAS component. The antibacterial efficiency of chewing gum was tested by using total plate count (TPC) and colony forming unit (CFU). Nine hours were required to microflora to reach TPC/CFU of before chewing gum consumption. This chewing gum not only provides mouth freshening activity but also helps in lowering dental decay, bad breath, and enamel whitening.

Keywords: colony forming unit (CFU), chewing gum, generally recognized as safe (GRAS), microbial growth, microorganisms, oral health, RACC, total plate count (TPC), antimicrobial agent, enamel whitening, oral pH

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Authors: Maryam Zahedifard, Fadhil Lafta Faraj, Nazia Abdul Majid, Hapipah Mohd Ali, Mahmood Ameen Abdulla

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The new quinazoline Schiff bases have been synthesized through condensation reaction of 2-aminobenzhydrazide with 5-bromosalicylaldehyde and 3-methoxy-5-bromosalicylaldehyde. The compound was investigated for anticancer activity against MCF-7 human breast cancer cell line. It demonstrated a remarkable antiproliferative effect, with an IC50 value of 3.41±0.34, after 72 hours of treatment. Most apoptosis morphological features in treated MCF-7 cells were observed by AO/PI staining. The results of cell cycle analysis indicate that compounds did not induce S and M phase arrest in cell after 24 hours of treatment. Furthermore, MCF-7 cells treated with compound subjected to apoptosis death, as exhibited by perturbation of mitochondrial membrane potential and cytochrome C release as well as increase in ROS generation. We also found activation of caspases 3/7 and -9. Moreover, acute toxicity results demonstrated the nontoxic nature of the compounds in mice. Our results showed the selected compound significantly induce apoptosis in MCF-7 cells via intrinsic pathway, which might be considered as a potential candidate for further in vivo and clinical breast cancer studies.

Keywords: quinazoline Schiff base, apoptosis, MCF-7, caspase, cell cycle, acute toxicity

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Authors: Alemu Tadesse Feroche

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In this study, some 2, 3 distributed quinazoline -4 (3H) - one derivative were synthesized using a three-step synthetic route. They were obtained in a good yield (59.5-85%) by applying different chemical reactions like cyclization and condensation reactions. The chemical structure of the final compounds was also verified by spectroscopic methods (IR, ¹HNMR) and elemental microanalysis. The in vivo antimalarial activity of these compounds on P. berghei infected mice was found to be moderate to high at an oral dose of 0.04846 mmol/kg /day. This is equal to 25 mg/kg of chloroquine phosphate, which causes 100% inhibition of the parasite. It is worth mentioning that most active compounds (E) -3 Phenyl -2- [2- (pyridine -4- yl) vinyl] -4 (3H) -quinazolinone IVa (64.02%, (E)-2-[2-(4 - Hydroxy-3 - methoxystyryl) - vinyl) -3 - phenyl -4 (3H ) - quinazolinone IVc (77.25%) and (E)-2 –[2 –(Pyridin -4-yl) –vinyl] -3 phenenylamine -4(3H) quinazolinone IVe (73.54%) showed a dose-dependent increase in present suppression in antimalarial activities. Furthermore, the synthesized compounds were screened for their in vitro antileishmanial activity against L. aethiopica isolate (CL/039/09). All tested compounds (IVa (0.03766 ug/ml), IVb (0.00538 ug/ml, IVc (0.00412 ug/ml, IVd (0.00110 ug/ml), IVe (0.03017 ug/ml) and IVf (0.03894 ug/ml)) showed excellent potency that is much better than amphotericin B (IC50 = 0,04359 ug/ml). The results of acute toxicity indicated that all test compounds (IVa –IVf) proved to be nontoxic and well tolerated by the experimental animals up to 300 mg/kg in oral and 140 mg/kg in parental studies.

Keywords: 4(3H)-quinazolinone, in vivo antimalarial activity, in vitro antileishmanial activity, acute toxicity

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Authors: Komal Saini, S. K. Sahoo, B. S. Bajwa

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The Laser Fluorimetery Technique has been used for the microanalysis of uranium content in water samples collected from different sources like the hand pumps, tube wells in the drinking water samples of SW & NE Punjab, India. The geographic location of the study region in NE Punjab is between latitude 31.21º- 32.05º N and longitude 75.60º-76.14º E and for SW Punjab is between latitude 29.66º-30.48º N and longitude 74.69º-75.54º E. The purpose of this study was mainly to investigate the uranium concentration levels of ground water being used for drinking purposes and to determine its health effects, if any, to the local population of these regions. In the present study 131 samples of drinking water collected from different villages of SW and 95 samples from NE, Punjab state, India have been analyzed for chemical and radiological toxicity. In the present investigation, uranium content in water samples of SW Punjab ranges from 0.13 to 908 μgL−1 with an average of 82.1 μgL−1 whereas in samples collected from NE- Punjab, it ranges from 0 to 28.2 μgL−1 with an average of 4.84 μgL−1. Thus, revealing that in the SW- Punjab 54 % of drinking water samples have uranium concentration higher than international recommended limit of 30 µgl-1 (WHO, 2011) while 35 % of samples exceeds the threshold of 60 µgl-1 recommended by our national regulatory authority of Atomic Energy Regulatory Board (AERB), Department of Atomic Energy, India, 2004. On the other hand in the NE-Punjab region, none of the observed water sample has uranium content above the national/international recommendations. The observed radiological risk in terms of excess cancer risk ranges from 3.64x10-7 to 2.54x10-3 for SW-Punjab, whereas for NE region it ranges from 0 to 7.89x10-5. The chemical toxic effect in terms of Life-time average Daily Dose (LDD) and Hazard Quotient (HQ) have also been calculated. The LDD for SW-Punjab varies from 0.0098 to 68.46 with an average of 6.18 µg/ kg/day whereas for NE region it varies from 0 to 2.13 with average 0.365 µg/ kg/day, thus indicating presence of chemical toxicity in SW Punjab as 35% of the observed samples in the SW Punjab are above the recommendation limit of 4.53 µg/ kg/day given by AERB for 60 µgl-1 of uranium. Maximum & Minimum values for hazard quotient for SW Punjab is 0.002 & 15.11 with average 1.36 which is considerably high as compared to safe limit i.e. 1. But for NE Punjab HQ varies from 0 to 0.47. The possible sources of high uranium observed in the SW- Punjab will also be discussed.

Keywords: uranium, groundwater, radiological and chemical toxicity, Punjab, India

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Authors: P. H. Mfengwana, S. S. Mashele, L. Verschaeve, R. Anthonissen, I. T. Manduna

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Gunnera perpensa is traditionally used mostly by women for the treatment of different gynaecological related conditions due to its proven uterine contractility effects. The uses of this plant include menstrual pain relief, treatment of infertility and promotion of easy labour. However, even though this plant species has been reported to possess numerous medicinal properties, to author’s best knowledge, its safety has not been investigated. Thus, this study was aimed at investigating the genotoxicity and antigenotoxicity of Gunnera perpensa aqueous, methanol and dichloromethane extracts. The in vitro toxicity of the plant extracts was assessed with the neutral red uptake (NRU) test. Genotoxic and antigenotoxic properties of Gunnera perpensa were investigated using high-throughput assays: bacterial Vitotox test and the alkaline comet assay with and without S9 activation on human C3A cells. Ethyl Methanesulfonate (EMS) and 4-nitroquinoline-oxide (4-NQO) were used as positive controls, respectively. All extracts showed toxicity in a dose-dependent manner; however, that does not mean they were all genotoxic. Methanol extract did show genotoxicity with S9 (metabolism) only at the highest concentration of 500 µg/ml due to increased DNA damage observed, however, no genotoxicity was observed from other concentrations. Therefore, the results show that Gunnera perpensa extracts are genotoxic and not safe for human use.

Keywords: antigenotoxicity, comet test, genotoxicity, Gunnera perpensa, vitotox assay

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Authors: Michela Terlizzi, Chiara Colarusso, Aldo Pinto, Rosalinda Sorrentino

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Sphingosine-1-phosphate (S1P) is a membrane-derived bioactive phospholipid exerting a multitude of effects on respiratory cell physiology and pathology through five S1P receptors (S1PR1-5). Higher levels of S1P have been registered in a broad range of respiratory diseases, including inflammatory disorders and cancer, although its exact role is still elusive. Based on our previous study in which we found that S1P/S1PR3 is involved in an inflammatory pattern via the activation of Toll-like Receptor 9 (TLR9), highly expressed on lung cancer cells, the main goal of the current study was to better understand the involvement of S1P/S1PR3 pathway/signaling during lung carcinogenesis, taking advantage of a mouse model of first-hand smoke exposure and of carcinogen-induced lung cancer. We used human samples of Non-Small Cell Lung Cancer (NSCLC), a mouse model of first-hand smoking, and of Benzo(a)pyrene (BaP)-induced tumor-bearing mice and A549 lung adenocarcinoma cells. We found that the intranuclear, but not the membrane, localization of S1PR3 was associated to the proliferation of lung adenocarcinoma cells, the mechanism that was correlated to human and mouse samples of smoke-exposure and carcinogen-induced lung cancer, which were characterized by higher utilization of S1P. Indeed, the inhibition of the membrane S1PR3 did not alter tumor cell proliferation after TLR9 activation. Instead, according to the nuclear localization of sphingosine kinase (SPHK) II, the enzyme responsible for the catalysis of the S1P last step synthesis, the inhibition of the kinase completely blocked the endogenous S1P-induced tumor cell proliferation. These results prove that the endogenous TLR9-induced S1P can on one side favor pro-inflammatory mechanisms in the tumor microenvironment via the activation of cell surface receptors, but on the other tumor progression via the nuclear S1PR3/SPHK II axis, highlighting a novel molecular mechanism that identifies S1P as one of the crucial mediators for lung carcinogenesis-associated inflammatory processes and that could provide differential therapeutic approaches especially in non-responsive lung cancer patients.

Keywords: sphingosine-1-phosphate (S1P), S1P Receptor 3 (S1PR3), smoking-mice, lung inflammation, lung cancer

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Authors: Antonina A. Shumakova, Sergey A. Khotimchenko

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The relevance of research is associated, on the one hand, with an ever-increasing volume of production and the expansion of the scope of application of engineered nanomaterials (ENMs), and on the other hand, with the lack of sufficient scientific information on the nature of the interactions of nanoparticles (NPs) with components of biogenic and abiogenic origin. In particular, studying the effect of ENMs (TiO2 NPs, SiO2 NPs, Al2O3 NPs, fullerenol) on the toxicometric characteristics of common contaminants such as lead and cadmium is an important hygienic task, given the high probability of their joint presence in food products. Data were obtained characterizing a multidirectional change in the toxicity of model toxicants when they are co-administered with various types of ENMs. One explanation for this fact is the difference in the adsorption capacity of ENMs, which was further studied in in vitro studies. For this, a method was proposed based on in vitro modeling of conditions simulating the environment of the small intestine. It should be noted that the obtained data are in good agreement with the results of in vivo experiments: - with the combined administration of lead and TiO2 NPs, there were no significant changes in the accumulation of lead in rat liver; in other organs (kidneys, spleen, testes and brain), the lead content was lower than in animals of the control group; - studying the combined effect of lead and Al2O3 NPs, a multiple and significant increase in the accumulation of lead in rat liver was observed with an increase in the dose of Al2O3 NPs. For other organs, the introduction of various doses of Al2O3 NPs did not significantly affect the bioaccumulation of lead; - with the combined administration of lead and SiO2 NPs in different doses, there was no increase in lead accumulation in all studied organs. Based on the data obtained, it can be assumed that at least three scenarios of the combined effects of ENMs and chemical contaminants on the body: - ENMs quite firmly bind contaminants in the gastrointestinal tract and such a complex becomes inaccessible (or inaccessible) for absorption; in this case, it can be expected that the toxicity of both ENMs and contaminants will decrease; - the complex formed in the gastrointestinal tract has partial solubility and can penetrate biological membranes and / or physiological barriers of the body; in this case, ENMs can play the role of a kind of conductor for contaminants and, thus, their penetration into the internal environment of the body increases, thereby increasing the toxicity of contaminants; - ENMs and contaminants do not interact with each other in any way, therefore the toxicity of each of them is determined only by its quantity and does not depend on the quantity of another component. Authors hypothesized that the degree of adsorption of various elements on the surface of ENMs may be a unique characteristic of their action, allowing a more accurate understanding of the processes occurring in a living organism.

Keywords: absorption, cadmium, engineered nanomaterials, lead

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Authors: Kayla Fanning

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Theocritus’ use of near-pastoral motifs in creating the lamenting narrative in “The Sorceress” idyll suggests a link between Simaetha and the quintessential shepherd that ultimately transcends the bucolic serenading structure, evident in “The Serenade”, as it pertains to depictions of women. In Theocritus’ “The Serenade”, an anonymous goatherd serenades his beloved, Amaryllis, in hopes of persuading her to reciprocate his love. This serenade soon turns into a vicious lament where all hope for reciprocation dissolves, leaving the goatherd severely melancholic and malignant. In “The Cyclops’ Serenade”, the cyclops, Polyphemus, sings of Galatea in solitude; in so doing, he negotiates between feelings of heartache and anger that eventually subside. His depiction of Galatea, in being less vindictive than the goatherd’s, manifests less toxicity. In adopting, and essentially creating, this serenading structure, Theocritus illustrates his ability to alter portrayals of women while maintaining the premise of the pastoral serenade; that is the shepherd's lament of his indifferent beloved. A thematic intertextual analysis of the idylls reveals a variety of ways in which the toxicity of the goatherd’s relation to Amaryllis is mutated or even inverted. In “The Sorceress”, a powerful witch named Simaetha spellbinds her unfaithful lover and angrily laments his betrayal in a way that is reminiscent of the goatherd's harmful behavior towards Amaryllis.

Keywords: femininity, pastoral, serenade, Theocritus

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Authors: Syeda Hira, Muhammad Gulfraz

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Background: Liver diseases cause serious health issues. Plants contain active compounds that significantly help in the treatment of various diseases. Ficus carica is traditionally used for the treatment of liver diseases. The purpose of the present study was the isolation and identification of active components from F.carica leaves which are responsible for hepatoprotective activity. Methods: The study was designed to identify the most active hepatoprotective sub-fraction from ethyl acetate fraction of Ficus carica by in vitro study and evaluation of its in vivo hepatoprotective effect in animal models. Ethyl acetate fraction was subjected to column, and a total of eight sub-fractions were obtained. In vitro, the hepatoprotective effect of all sub-fractions was determined on HepG2 cell lines. Toxicity was induced by CCl₄ (Carbon tetrachloride), and silymarin was used as a positive control. On the basis of the results, the most active sub-fraction was subjected to LC-MS and FT-IR analysis for the identification of bioactive compounds. In vivo, the hepatoprotective effect was determined in mice. Toxicity was induced by CCl₄; at the end of the experiment, biochemical parameters such as ALT, AST, ALP, bilirubin, and total protein were estimated in serum. Histopathology of liver tissues was also done. Results: Sub-fraction FVI exhibited significant (P<0.05) hepatoprotective activity as compared to other sub-fractions, which was almost similar to the standard drug silymarin. Six known bioactive compounds were identified from this sub-fraction after LC-MS analysis. In vivo, the hepatoprotective activity of sub-fraction FVI was evaluated in CCl₄-induced toxicated mice. Administration of CCl₄ significantly increased level of ALT (Alanine transaminase), AST (Aspartate aminotransferase), ALP (Alkaline phosphatase), and bilirubin and decreased the total protein. Treatment with sub-fraction FVI significantly (p<0.05) reversed the level of these biomarkers toward normal at both doses of 25 mg/kg and 50 mg/kg. Conclusion: Our findings confirmed the hepatoprotective effect of ethyl acetate fraction of F.carica. It could be a good candidate for the development of a natural hepatoprotective drug; pre-clinical investigation on ethyl acetate fraction is recommended.

Keywords: Ficus carica, hepatoprotective, CCl₄, bioactive compounds, liver markers

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Authors: Tomas Makaras

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Sticklebacks (Gasterosteiformes) are increasingly used in ecological and evolutionary research and become well-established role as model species for biologists. However, ecotoxicology studies concerning behavioural effects in sticklebacks regarding stress responses, mainly induced by chemical mixtures, have hardly been addressed. Moreover, although many authors in their studies emphasised the similarity between three-spined and nine-spined stickleback in morphological, neuroanatomical and behavioural adaptations to environmental changes, several comparative studies have revealed considerable differences between these species in and their susceptibility and resistance to variousstressors in laboratory experiments. The hypothesis of this study was that three-spined and nine-spined stickleback species will demonstrate apparent differences in response patterns and sensitivity to metal-based chemicals stimuli. For this purpose, we investigated the swimming behaviour (including mortality rate based on 96-h LC50 values) of two ecologically similar three-spined (Gasterosteusaculeatus) and nine-spined sticklebacks (Pungitiuspungitius) to short-term (up to 24 h) metal mixture (MIX) exposure. We evaluated the relevance and efficacy of behavioural responses of test species in the early toxicity assessment of chemical mixtures. Fish exposed to six (Zn, Pb, Cd, Cu, Ni and Cr) metals in the mixture were either singled out by the Water Framework Directive as priority or as relevant substances in surface water, which was prepared according to the environmental quality standards (EQSs) of these metals set for inland waters in the European Union (EU) (Directive 2013/39/EU). Based on acute toxicity results, G. aculeatus found to be slightly (1.4-fold) more tolerant of MIX impact than those of P. pungitius specimens. The performed behavioural analysis showed the main effect on the interaction between time, species and treatment variables. Although both species exposed to MIX revealed a decreasing tendency in swimming activity, these species’ responsiveness to MIX was somewhat different. Substantial changes in the activity of G. aculeatus were established after 3-h exposure to MIX solutions, which was 1.43-fold lower, while in the case of P. pungitius, 1.96-fold higher than established 96-h LC50 values for each species. This study demonstrated species-specific differences in response sensitivity to metal-based water pollution, indicating behavioural insensitivity of P. pungitiuscompared to G. aculeatus. While many studies highlight the usefulness and suitability of nine-spined sticklebacks for evolutionary and ecological research, attested by their increasing popularity in these fields, great caution must be exercised when using them as model species in ecotoxicological research to probe metal contamination. Meanwhile, G. aculeatus showed to be a promising bioindicator species in the environmental ecotoxicology field.

Keywords: acute toxicity, comparative behaviour, metal mixture, swimming activity

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Authors: Sudhir Kumar, V. R. Sinha

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Background: The topical and systemic toxicity associated with present nonmelanoma skin cancer (NMSC) treatment therapy using 5-Fluorouracil (5-FU) make it necessary to develop a novel delivery system having lesser toxicity and better control over drug release. Solid lipid nanoparticles offer many advantages like: controlled and localized release of entrapped actives, nontoxicity, and better tolerance. Aim:-To investigate safety and efficacy of 5-FU loaded solid lipid nanoparticles as a topical delivery system for the treatment of nonmelanoma skin cancer. Method: Topical solid lipid nanoparticles of 5-FU were prepared using Compritol 888 ATO (Glyceryl behenate) as lipid component and pluronic F68 (Poloxamer 188), Tween 80 (Polysorbate 80), Tyloxapol (4-(1,1,3,3-Tetramethylbutyl) phenol polymer with formaldehyde and oxirane) as surfactants. The SLNs were prepared with emulsification method. Different formulation parameters viz. type and ratio of surfactant, ratio of lipid and ratio of surfactant:lipid were investigated on particle size and drug entrapment efficiency. Results: Characterization of SLNs like–Transmission Electron Microscopy (TEM), Differential Scannig calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), Particle size determination, Polydispersity index, Entrapment efficiency, Drug loading, ex vivo skin permeation and skin retention studies, skin irritation and histopathology studies were performed. TEM results showed that shape of SLNs was spherical with size range 200-500nm. Higher encapsulation efficiency was obtained for batches having higher concentration of surfactant and lipid. It was found maximum 64.3% for SLN-6 batch with size of 400.1±9.22 nm and PDI 0.221±0.031. Optimized SLN batches and marketed 5-FU cream were compared for flux across rat skin and skin drug retention. The lesser flux and higher skin retention was obtained for SLN formulation in comparison to topical 5-FU cream, which ensures less systemic toxicity and better control of drug release across skin. Chronic skin irritation studies lacks serious erythema or inflammation and histopathology studies showed no significant change in physiology of epidermal layers of rat skin. So, these studies suggest that the optimized SLN formulation is efficient then marketed cream and safer for long term NMSC treatment regimens. Conclusion: Topical and systemic toxicity associated with long-term use of 5-FU, in the treatment of NMSC, can be minimized with its controlled release with significant drug retention with minimal flux across skin. The study may provide a better alternate for effective NMSC treatment.

Keywords: 5-FU, topical formulation, solid lipid nanoparticles, non melanoma skin cancer

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Authors: Roberta Pecoraro, Elena Maria Scalisi

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Engineered Nanoparticles (ENPs) and Nanomaterials (ENMs) concern an active research area and a sector in full expansion. They have physical-chemical characteristics and small size that improve their performance compared to common materials. Due to the increase in their production and their subsequent release into the environment, new strategies are emerging to assess risk of nanomaterials. NPs can be released into the environment through aquatic systems by human activities and exert toxicity on living organisms. We evaluated the potential toxic effect of cerium oxide (CeO2) nanoparticles because it’s used in different fields due to its peculiar properties. In order to assess nanoparticles toxicity, Fish Embryo Toxicity (FET) test was performed. Powders of CeO2 NPs supplied by the CNR-IMM of Catania are indicated as CeO2 type 1 (as-prepared) and CeO2 type 2 (modified), while CeO2 type 3 (commercial) is supplied by Sigma-Aldrich. Starting from a stock solution (0.001g/10 ml dilution water) of each type of CeO2 NPs, the other concentration solutions were obtained adding 1 ml of the stock solution to 9 ml of dilution water, leading to three different solutions of concentration (10-4, 10-5, 10-6 g/ml). All the solutions have been sonicated to avoid natural tendency of NPs to aggregate and sediment. FET test was performed according to the OECD guidelines for testing chemicals using our internal protocol procedure. A number of eight selected fertilized eggs were placed in each becher filled with 5 ml of each concentration of the three types of CeO2 NPs; control samples were incubated only with dilution water. Replication was performed for each concentration. During the exposure period, we observed four endpoints (embryo coagulation, lack of formation of somites, failure to lift the yolk bag, no heartbeat) by a stereomicroscope every 24 hours. Immunohistochemical analysis on treated larvae was performed to evaluate the expression of metallothioneins (MTs), Heat Shock Proteins 70 (HSP70) and 7-ethoxyresorufin-O-diethylase (EROD). Our results have not shown evident alterations on embryonic development because all embryos completed the development and the hatching of the eggs, started around the 48th hour after exposure, took place within the last observation at 72 hours. A good reactivity, both in the embryos and in the newly hatched larvae, was found. The presence of heartbeat has also been observed in embryos with reduced mobility confirming their viability. A higher expression of EROD biomarker was observed in the larvae exposed to the three types of CeO2, showing a clear difference with the control. A weak positivity was found for MTs biomarker in treated larvae as well as in the control. HSP70 are expressed homogeneously in all the type of nanoparticles tested but not too much greater than control. Our results are in agreement with other studies in the literature, in which the exposure of Danio rerio larvae to other metal oxide nanoparticles does not show adverse effects on survival and hatching time. Further studies are necessary to clarify the role of these NPs and also to solve conflicting opinions.

Keywords: Danio rerio, endpoints, fish embryo toxicity test, metallic nanoparticles

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Authors: F. Haddadj, F. Acheuk, S. Hamdi, S. Zenia, A. Smai, H. Saadi, B. Doumandji-Mitiche

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Locust is causing significant losses in agricultural production in the countries concerned by the invasion. Up to the present control strategy has consisted only of the spreaders chemicals; they have proven harmful to the environment and. For this, a new control method appeared it comes to the biological control based mostly by using microorganism. It is in that sense is we've made our contribution by the use of a biopesticide which is entomopathogenic fungus Metarhizium anisopliae var acridium "Green Muscle" on part of the cuticule the larval of fifth instar locust Schistocerca gregaria (Forskål, 1775). Preliminary test on the study of the pathogenicity of M. anisopliae var acridium biocontrol agent, was conducted in the laboratory on L5 S. gregaria, on which we inoculated treatment in the digestive tract and it administrant 20μl of entomopathogenic solution orally at a dose DL50 = 3.25 x107 sp./ ml (median lethal dose estimated at earlier), 5 days after treatment individuals are sacrificed. After dissection cuticles are recovered and then subjected to histological sections. The histological technique followed is that of Martoja Martoja-Pierson (1967). Microscopic observation revealed alterations in the architecture of the cuticule which leads to disorganization of cell layers.

Keywords: biopesticide, cuticle, desert locust, toxicity

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