Involvement of mitochondrial-mediated caspase-3 activation and lysosomal labilization in acrylamide-induced liver toxicity

Acrylamide (ACR) is a chemical frequently used in both industrial and synthetic processes and may be produced during food processing. ACR at very high concentrations is postulated to exert its toxicity through the stimulation of an oxidative stress. ACR in excessive doses induces the central nervous system, reproduction, and genetic toxicity. However, ACR effects on the liver, a major organ of drug metabolism, have not been adequately explored. In addition, the role of mitochondria in an ACR-mediated hepatotoxicity is still unclear. The aim of this study was to investigate the cytotoxic mechanisms attributed to ACR using isolated rat hepatocytes. Hepatocytes were isolated by the collagenase perfusion method and incubated with an EC50_2hr concentration of ACR for 3 hr. The EC50_{2 hr} of ACR on isolated rat hepatocytes was determined to be 1 mM. Based on our results, hepatocytes cytotoxicity of ACR (1 mM) was mediated by a reactive oxygen species formation and lipid peroxidation. Incubation of hepatocytes with ACR produced rapid hepatocyte glutathione depletion which is another marker of the cellular oxidative stress. ACR cytotoxicity was also associated with mitochondrial injury as evidenced by the decline of mitochondrial membrane potential and lysosomal membrane leakiness. Our results also showed that ACR induced caspase-3 activation, the final mediator of apoptosis signaling. These findings contribute to a better understanding underlying mechanisms involved in ACR hepatotoxicity originating from the oxidative stress and ending in mitochondrial/lysosomal damage and cell death signaling.     

Aerobic treatment of palm oil mill effluent

In this study treatment of palm oil mill effluent (POME) was investigated using aerobic oxidation based on an activated sludge process. The effects of sludge volume index, scum index and mixed liquor suspended solids during the acclimatizing phase and biomass build-up phase were investigated in order to ascertain the reactor stability. The efficiency of the activated sludge process was evaluated by treating anaerobically digested and diluted raw POME obtained from Golden Hope Plantations, Malaysia. The treatment of POME was carried out at a fixed biomass concentration of 3900+/-200mg/L, whereas the corresponding sludge volume index was found to be around 105+/-5mL/g. The initial studies on the efficiency of the activated sludge reactor were carried out using diluted raw POME for varying the hydraulic retention time, viz: 18, 24, 30 and 36h and influent COD concentration, viz: 1000, 2000, 3000, 4000 and 5000mg/L, respectively. The results showed that at the end of 36h of hydraulic retention time for the above said influent COD, the COD removal efficiencies were found to be 83%, 72%, 64%, 54% and 42% whereas at 24h hydraulic retention time they were 57%, 45%, 38%, 30% and 27%, respectively. The effectiveness of aerobic oxidation was also compared between anaerobically digested and diluted raw POME having corresponding CODs of 3908 and 3925mg/L, for varying hydraulic retention time, viz: 18, 24, 30, 36, 42, 48, 54 and 60h. The dissolved oxygen concentration and pH in the activated sludge reactor were found to be 1.8-2.2mg/L and 7-8.5, respectively. The scum index was found to rise from 0.5% to 1.9% during the acclimatizing phase and biomass build-up phase.     

Sorption of ametryn and imazethapyr in twenty-five soils from Pakistan and Australia.

Sorption of ametryn and imazethapyr in 25 soils from Pakistan and Australia was investigated using the batch method. The soils varied widely in their intrinsic capacities to sorb these herbicides as shown by the sorption coefficients, Kd, which ranged from 0.59 to 47.6 for ametryn and 0.02 to 6.94 for imazethapyr. Generally the alkaline soils of Pakistan had much lower Kd values of both herbicides than the soils of Australia. Both soil pH and soil organic carbon (SOC) were correlated significantly with the sorption of ametryn, whereas only soil pH was strongly correlated with imazethapyr sorption. No correlation was found between Kd values of the herbicides and the clay contents of the soils. Multiple regression analysis showed that Kd values were better correlated (r2=0.94 and 0.89 for ametryn and imazethapyr, respectively) if SOC and pH were simultaneously taken into account. The study indicated that sorption of these herbicides in the alkaline soils of Pakistan was low and consequently there is considerable risk of groundwater contamination.     

Determination of organochlorine and organophosphate pesticide residues in fruits, vegetables and sediments.

A method is described for the determination of organochlorine and organophosphate pesticide residues in fruits, vegetables and sediments. The concentrated solvent extract was sealed in a polymeric membrane tube, dialysed in cyclohexane and the solvent replaced with hexane. The organophosphates were analysed on a specific thermionic detector without further clean-up. For the organochlorine pesticides the extract was eluted through 3 g of alumina and analysed on GC/ECD. The clean-up for sediment extract was carried out on a 10 g alumina column with 100 mL hexane containing 5% acetone and the eluate was concentrated to 5 mL. The detection limit for organophosphates on a 40 g sample and a final volume of 10 mL was on the average 0.01 mg/kg. The detection limit for organochlorine pesticides, with the final volume of 25 mL, was 0.005 mg/kg for all pesticides except for p,p'-DDT and endosulfan sulphate, which was 0.01 mg/kg. The detection limit for organochlorine pesticides in sediment, with the final volume of 2 mL, was less than 1 microgram/kg and for organophosphate pesticides less than 10 micrograms/kg when the final volume was made to 0.5 mL. At the detection limits the method produced a very high coefficient of variation for both organochlorine and organophosphate pesticides.     

Cleanup of biological extracts by a Pasteur pipette column and a comparison of in vivo-accumulated chlorinated pesticide residues with other clean up systems

Biological samples are extracted with n-hexane/acetone (60:40) and 1 mL of the concentrated extract is eluted through a pasteur pipette column prepacked with alumina (0.3 g) and silicic acid (0.25 g) with 10 mL n-hexane (containing 4% acetone). The fat and other co-extractives are retained by the column and clear eluate is directly injected on a GLC column for determination on electron capture detector. A comparison of the Pasteur pipette cleanup with the modified method of Cole et al. (1967) on 41 samples of fish, One Step Method, (Ahmad and Marolt (1986] on 86 samples of fish and Maunder et al. (1964) on 10 type of wildlife (100 samples) was made. The Pasteur pipette method gives results which are significantly higher (p greater than 0.5) than the other methods except the One step method. The Pasteur pipette method has a detection limit of 0.01 microgram/g for DDT and its metabolites.     

Acute toxicity of two insecticides to tubificid worms, Tubifex tubifex and Limnodrilus hoffmeisteri

Acute toxicity and presumable harmless concentration of two commercial insecticides, Furadan 3G and Malataf 50E were reported for tubificid worms, Tubifex tubifex and Limnodrilus hoffmeisteri. L. hoffmeisteri were found more susceptible to both the insecticides; the Furadan was found to be more toxic than Malataf to tubificid worms. Analysis of variants showed a significant difference between safe concentrations of the same insecticide to different species.     

Used lubricating oil recycling using hydrocarbon solvents

A solvent extraction process using new hydrocarbon solvents was employed to treat used lubricant oil. The solvents used were liquefied petroleum gas (LPG) condensate and stabilized condensate. A demulsifier was used to enhance the treatment process. The extraction process using stabilized condensate demonstrated characteristics that make it competitive with existing used oil treatment technologies. The process is able to reduce the asphaltene content of the treated lubricating oil to 0.106% (w/w), the ash content to 0.108%, and the carbon residue to 0.315% with very low levels of contaminant metals. The overall yield of oil is 79%. The treated used oil can be recycled as base lubricating oil. The major disadvantage of this work is the high temperature of solvent recovery. Experimental work and results are presented in detail.