In operant conditioning experiments, two methods are commonly used to motivate laboratory rats to perform designated tasks. The first is restricting food so that rats are forced to lose 20% of body weight within one week, followed by maintenance at 80% of the baseline weight for the remainder of the experiment. The second is restricting access to water to 15 min in each 24 h period. These methods are effective in motivating the animals. There is, however, little information available on the effects on performance in tests of behaviour that are not related to operant conditioning. In addition, it is not clear if these commonly used methods of food and water restriction will lead to physiological stress as indicated by an elevation of serum corticosterone. Male rats were either food-restricted to reduce and maintain their weight at 80% of baseline weight, or were restricted to 15 min access to water every 24 h. Activity in the open field was significantly greater in food-restricted rats than in water-restricted or control rats, but freezing behaviour was similar in all experimental groups. Food-restricted rats had a higher mean serum corticosterone level than water-restricted and control rats 37 days after the start of the experimental period. These data suggested that chronically restricting food and maintenance of body weight at 80% of baseline body weight led to significant behavioural changes and physiological stress. In contrast, water restriction did not lead to changes in behaviour or corticosterone levels. A second experiment was conducted to compare the effects of food restriction to 80% of baseline body weight, as described above, with a less stringent protocol in which test rats were initially reduced to 80% of baseline weight, but were then maintained at 80% of an ad libitum fed control rat's weight. Serum corticosterone levels and adrenal gland weights were measured after the initial week of forced weight loss and after maintenance for 21 days. Forced loss of 20% of body weight in the first week led to significantly increased serum corticosterone levels and adrenal gland weights compared to ad libitum fed controls. Serum corticosterone levels and adrenal gland weights in rats maintained at 80% of their initial body weight for 21 days remained higher than ad libitum fed control rats. However, rats maintained at 80% of an ad libitum fed control rat's weight did not differ from control rats in serum corticosterone levels or adrenal gland weights at the end of the 21-day study period. Adjustment of the feeding regimen in this manner eliminated physiological evidence of chronic stress.

Copper is an essential element and also produces adverse health consequences when overloaded. Food and water are the main sources of copper intake, however few studies have been conducted to investigate the difference between the ways of its intake in water and food in animals. In this study, copper was fed to mice with food as well as water (two groups: water and diet) for three months at concentrations of 6, 15 and 30 ppm. The copper concentration in water was adjusted for keeping the same amount during its intake in food. The experimental studies show a slow growth rate, lower hepatic reduced glutathione (GSH)/superoxide dismutase (SOD) activity and higher serum ‘free’ copper in the water group. The brain's soluble amyloid-beta 1-42 (Aβ₄₂) of the water group was significantly higher than that of the diet group at the levels of 6 and 15 ppm. In conclusion, copper in the water group significantly increased the soluble Aβ₄₂ in the brain and the ‘free’ copper in the serum, decreased the growth rate and hepatic GSH/SOD activity. The research studies carried out suggest that the copper in water is more ‘toxic’ than copper in diet and may increase the risk of Alzheimer's disease (AD).

Evaluation of acetylcholinesterase (AChE) activity and determination of organophosphorus pesticides (OPs) are of great importance for the clinical diagnosis of several serious diseases correlated with their variations in human blood serum. In this study, a highly selective and sensitive ratiometric fluorescent probe was innovatively fabricated for the evaluation of AChE activity and the determination of OPs in tap water and food on the basis of the inner filter effect (IFE) between nitrogen-doped carbon dots (N-CDs) and 2,3-diaminophenazine (DAP). N-CDs were synthesized via a one-pot hydrothermal method by using pancreatin and 1,2-ethanediamine as precursors. N-CDs showed excellent fluorescence properties and negligible cytotoxicity on human cervical carcinoma HeLa cells and human embryonic kidney 293T cells, suggesting their further biological applications. Upon the addition of AChE and choline oxidase, acetylcholine was catalyzed to produce choline that was further oxidized to produce H₂O₂. In the presence of horseradish peroxidase, o-phenylenediamine reacted with H₂O₂ to produce fluorescent DAP. Therefore, a ratiometric fluorescent probing platform existed via IFE between N-CDs with a fluorescence signal at 450 nm and DAP with a fluorescence signal at 574 nm. OPs irreversibly impeded the catalytic activity of AChE, finally leading to the decrease of DAP amount and the variation of ratiometric fluorescent signal. Under optimal conditions, such a fluorescent probe showed relatively low detection limits of 0.38 U/L for AChE, 3.2 ppb for dichlorvos, and 13 ppb for methyl-parathion. Practical application of this ratiometric fluorescent probe to detect OPs was further verified in tap water and food samples with satisfying results that were highly consisted with the results obtained by GC–MS.

Herein, the synthesis of bovine serum albumin-Cu(II) hybrid nanoflowers (BSA-NFs) through the building blocks of bovine serum albumin (BSA) and copper(II) ions in phosphate buffered saline (PBS) and their use as adsorbent for cadmium and lead ions are reported. The BSA-NFs, for the first time, were efficiently utilized as novel adsorbent for solid phase extraction (SPE) of cadmium and lead ions in water, food, cigarette and hair samples. The method is based on the separation and pre-concentration of Cd(II) and Pb(II) by BSA-NFs prior to determination by slurry analysis via flame atomic absorption spectrometry (FAAS). The analytes were adsorbed on BSA-NFs under the vortex mixing and then the ion-loaded slurry was separated and directly introduced into the flame AAS nebulizer by using a hand-made micro sample introduction system to eliminate a number of drawbacks. The effects of analytical key parameters, such as pH, amount of BSA-NFs, vortexing time, sample volume, and matrix effect of foreign ions on adsorbing of Cd(II) and Pb(II) were systematically investigated and optimized. The limits of detection (LODs) for Cd(II) and Pb(II) were calculated as 0.37 μg L−1 and 8.8 μg L−1, respectively. The relative standard deviation percentages (RSDs) (N = 5) for Cd(II) and Pb(II) were 7.2%, and 5.0%, respectively. The accuracy of the developed procedure was validated by the analysis of certified reference materials (TMDA-53.3 Fortified Water, TMDA-70 Fortified Water, SPS-WW2 Waste Water, NCSDC-73349 Bush Branches and Leaves) and by addition/recovery analysis. The quantitative recoveries were obtained for the analysis of certified reference materials and addition/recovery tests. The method was successfully applied to the analysis of cadmium and lead in water, food, cigarette and hair samples.