Monitoring of total type ii pyrethroid pesticides in citrus oils and water by converting to a common product 3-phenoxybenzoic acid

Pyrethroids are a class of insecticides that are becoming increasingly popular in agricultural and home use applications. Sensitive assays for pyrethroid insecticides in complex matrices are difficult with both instrumental and immunochemical methods. Environmental analysis of the pyrethroids by immunoassay requires either knowing which pyrethroids contaminate the source or the use of nonspecific antibodies with cross-reactivities to a class of compounds. We describe an alternative method that converts the type II pyrethroids to a common chemical product, 3-phenoxybenzoic acid (3-PBA), prior to analysis. This method is much more sensitive than detecting the parent compound, and it is much easier to detect a single compound rather than an entire class of compounds. This is useful in screening for pyrethroids as a class or in situations where a single type of pyrethroid is used. We demonstrated this technique in both citrus oils and environmental water samples with conversion rates of the pyrethroid to 3-PBA that range from 45 to 75% and methods that require no extraction steps for either the immunoassay or the liquid chromatography-tandem mass spectrometry (LC-MS/MS) techniques. Limits of detection for this technique applied to orange oil are 5 nM, 2 μM, and 0.8 μM when detected by LC-MS/MS, gas chromatography-mass spectrometry, and immunoassay, respectively. The limit of detection for pyrethroids in water when detected by immunoassay was 2 nM.     

Development of an immunoassay for the residues of the herbicide bensulfuron-methyl

To develop a competitive indirect enzyme-linked immunosorbent assay based on polyclonal antibodies for the detection of the sulfonylurea herbicide bensulfuron-methyl, seven structurally related haptens were synthesized. Four of them mimicking the target analyte were conjugated to keyhole limpet hemocyanin by the N-hydroxysuccinimide activated ester method to use as immunogens, and all of them were conjugated to bovine serum albumin to use as plate-coating antigens. Polyclonal antibodies raised in rabbits and the coating antigens were screened and selected for the assay in simple homologous and heterologous ELISA formats. Three sensitive heterologous ELISAs were selected and optimized, showing the average IC(50) values of bensulfuron-methyl as low as 0.17, 0.09, and 0.09 ng/mL, the detection ranges of 0.04-0.60, 0.01-0.60, and 0.04-0.25 ng/mL, and the lowest detection limits of 0.03, 0.002, and 0.03 ng/mL, respectively. The cross-reactivities of other sulfonylurea herbicides and metabolites of bensulfuron-methyl to the antibodies were less than 15% in the two assays. Recoveries from the analyte-fortified water samples in assay I were in the range of 81-125% by simple dilution. The correlation between the ELISA and HPLC was 0.999 (n = 15) with a slope of 1.37 in the analysis of groundwater samples fortified with bensulfuron-methyl. The results obtained strongly indicate that the ELISA can be a highly sensitive and convenient tool for detecting bensulfuron-methyl residues in agricultural and environmental samples.     

Changes in neighbourhood food store environment, food behaviour and body mass index, 1981--1990

OBJECTIVE: This paper examines trends in the neighbourhood food store environment (defined by the number and geographic density of food stores of each type in a neighbourhood), and in food consumption behaviour and overweight risk of 5779 men and women. DESIGN: The study used data gathered by the Stanford Heart Disease Prevention Program in four cross-sectional surveys conducted from 1981 to 1990. SETTING: Four mid-sized cities in agricultural regions of California. SUBJECTS: In total, 3154 women and 2625 men, aged 25-74 years. RESULTS: From 1981 to 1990, there were large increases in the number and density of neighbourhood stores selling sweets, pizza stores, small grocery stores and fast-food restaurants. During this period, the percentage of women and men who adopted healthy food behaviours increased but so did the percentage who adopted less healthy food behaviours. The percentage who were obese increased by 28% in women and 24% in men.ConclusionFindings point to increases in neighbourhood food stores that generally offer mostly unhealthy foods, and also to the importance of examining other food pattern changes that may have a substantial impact on obesity, such as large increases in portion sizes during the 1980s.     

Headspace oxygen in sample vials affects volatiles production of meat during the automated purge-and-Trap/GC analyses.

Headspace oxygen in sample vial for the purge-and-trap dynamic headspace/gas chromatography method oxidizes meat if held hours before purging, influences volatile profiles, and misrepresents the true composition of volatiles. Helium flush and helium flush plus oxygen absorber were used to eliminate residual oxygen and minimize oxidative changes in meat during sample holding time. Both helium flush and helium flush plus oxygen absorber treatments were effective in preventing an increase in 2-thiobarbituric acid reactive substances (TBARSs) and volatiles production in raw meat for up to 640 min of sample holding. With helium flush plus oxygen absorber, only 1-octen-3-ol increased during the 1280-min sample holding time. However, the hexanal peak in raw meat was interfered by 2,6-dimethyl heptane when oxygen absorber was added. Therefore, use of oxygen absorber was not appropriate for raw meat. Helium flush reduced oxidative changes in cooked meat during sample holding time but was not able to stop oxidative changes in meat after 160 min sample holding. A combination of helium flush and oxygen absorber was effective in preventing volatiles production in cooked meat for over 20 h of sample holding at 4 degrees C.     

Hapten and antibody production for a sensitive immunoassay determining a human urinary metabolite of the pyrethroid insecticide permethrin

Permethrin is the most popular synthetic pyrethroid insecticide in agriculture and public health. For the development of the enzyme-linked immunosorbent assay (ELISA) to evaluate human exposure to permethrin, the glycine conjugate (DCCA-glycine) of a major metabolite, cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), of permethrin was established as the target analyte. Four different types of the cis- and trans-isomers of immunizing haptens were synthesized as follows: N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine (hapten 3), N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)-4-amino-l-phenylalanine (hapten 5), N-(N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine)amino-6-(2,4-dinitrophenyl)aminohexanoic acid (hapten 9), and N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine-4-oxobutanoic acid (hapten 24). Sixteen polyclonal antibodies produced against each cis- or trans-hapten-thyroglobulin conjugate as immunogens were screened against numerous hapten-bovine serum albumin conjugates as coating antigens. Six ELISAs with both a heterologous hapten structure and a heterologous hapten configuration (cis/trans or trans/cis) between antibody and coating antigen showed a high sensitivity for the target analyte. The IC50 was 1.3, 2.1, and 2.2 microg/L for the trans-target analyte and 0.4, 2.3, and 2.8 microg/L for the cis-target analyte. The immunizing haptens, except for hapten 5, provided the target specific antibodies. Molecular modeling of the haptens supported the selection of reasonable immunizing haptens that best mimicked the target analyte. Hapten 5 was suitable as a coating antigen rather than as an immunogen since it had a different geometry. Very low cross-reactivities were measured to permethrin, its free metabolite (DCCA), PBA-glycine conjugate, and glycine. The ELISA will be optimized for the detection of total cis/trans-DCCA-glycine in human urine samples.     

Astaxanthin Reduces Stemness Markers in BT20 and T47D Breast Cancer Stem Cells by Inhibiting Expression of Pontin and Mutant p53

Astaxanthin (AST) is a product made from marine organisms that has been used as an anti-cancer supplement. It reduces pontin expression and induces apoptosis in SKBR3, a breast cancer cell line. Using Western blotting and qRT-PCR analyses, this study revealed that in the T47D and BT20 breast cancer cell lines, AST inhibits expression of pontin and mutp53, as well as the Oct4 and Nanog cancer stem cell (CSC) stemness genes. In addition, we explored the mechanism by which AST eradicates breast cancer cells using pontin siRNAs. Pontin knockdown by pontin siRNA reduced proliferation, Oct4 and Nanog expression, colony and spheroid formation, and migration and invasion abilities in breast cancer cells. In addition, reductions in Oct4, Nanog, and mutp53 expression following rottlerin treatment confirmed the role of pontin in these cells. Therefore, pontin may play a central role in the regulation of CSC properties and in cell proliferation following AST treatment. Taken together, these findings demonstrate that AST can repress CSC stemness genes in breast cancer cells, which implies that AST therapy could be used to improve the efficacy of other anti-cancer therapies against breast cancer cells.     

Effect of enzyme treatment and wood pulp variation on physical characteristics and fabric hand of lyocell fabrics

The purpose of the research was to examine the effect of three different levels of enzyme treatment on the physical characteristics and the end-use suitability of the lyocell fabrics made with four different types of commercial wood pulp. The appropriate enzyme concentration for obtaining an optimum as well as consistent physical characteristics and fabric hand trait was 3 g/l for the concentration levels tested in the present investigation. Weight loss was more affected by higher enzyme concentration than other physical properties. H2 was least affected by enzyme treatment for all three physical properties and fabric hand. 5 g/l concentration exhibited little difference from 3 g/l in the physical characteristics, whereas the KES-FB values indicated a significant loss of fabric hand in most PHVs with the 5 g/l concentration level. Among different garment parameters, all four fabric types were relatively inappropriate for the men’s slacks (MS) fabric due to the lower hand value of koshi required by the MS parameter. However, despite the relatively low koshi values, high fukurami values required for men’s dress shirt (MWDS) resulted in the highest THV among the four garment parmeters. The four fabric types, which represent the usage of four different wood pulps, in general seem to exhibit a higher applicability to women’s winter thin dress (WWTD) than women’s winter suit (WWS) garment parameter.     

Astaxanthin Confers a Significant Attenuation of Hippocampal Neuronal Loss Induced by Severe Ischemia-Reperfusion Injury in Gerbils by Reducing Oxidative Stress

Astaxanthin is a powerful biological antioxidant and is naturally generated in a great variety of living organisms. Some studies have demonstrated the neuroprotective effects of ATX against ischemic brain injury in experimental animals. However, it is still unknown whether astaxanthin displays neuroprotective effects against severe ischemic brain injury induced by longer (severe) transient ischemia in the forebrain. The purpose of this study was to evaluate the neuroprotective effects of astaxanthin and its antioxidant activity in the hippocampus of gerbils subjected to 15-min transient forebrain ischemia, which led to the massive loss (death) of pyramidal cells located in hippocampal cornu Ammonis 1-3 (CA1-3) subfields. Astaxanthin (100 mg/kg) was administered once daily for three days before the induction of transient ischemia. Treatment with astaxanthin significantly attenuated the ischemia-induced loss of pyramidal cells in CA1-3. In addition, treatment with astaxanthin significantly reduced ischemia-induced oxidative DNA damage and lipid peroxidation in CA1-3 pyramidal cells. Moreover, the expression of the antioxidant enzymes superoxide dismutase (SOD1 and SOD2) in CA1-3 pyramidal cells were gradually and significantly reduced after ischemia. However, in astaxanthin-treated gerbils, the expression of SOD1 and SOD2 was significantly high compared to in-vehicle-treated gerbils before and after ischemia induction. Collectively, these findings indicate that pretreatment with astaxanthin could attenuate severe ischemic brain injury induced by 15-min transient forebrain ischemia, which may be closely associated with the decrease in oxidative stress due to astaxanthin pretreatment.