Effects of Maternal Exposure to Piperonyl Butoxide (PBO) on Behavioral Development in F1‐Generation Mice

Female mice were exposed maternally to piperonyl butoxide (PBO) through diet to provide dietary levels of 0% (control), 0.01%, 0.03%, and 0.09% during gestation and lactation periods, and selected reproductive and neurobehavioral parameters were measured in the F₁ generation. There was no adverse effect of PBO on litter size, litter weight, or sex ratio at birth. The average body weights of male offspring decreased significantly in dose‐related manners on postnatal days (PNDs) 0, 4, 7, and 14 (p = 0.0019, 0.0096, 0.033, and 0.038, respectively) during the lactation period. In female offspring, the average body weights decreased in dose‐related manners on PNDs 0, 4, 7, and 14 (p = 0.0027, 0.0104, 0.0193, and 0.0062, respectively). The survival of dams slightly decreased (p = 0.0209) in the high‐dose group during the lactation period. With respect to behavioral developmental parameters, surface righting on PND 7 of male and female offspring was delayed significantly in a dose‐related manner (p < 0.001 in each). Swimming direction on PND 7 of male offspring was delayed significantly in a dose‐related manner (p < 0.01), and for female offspring it was delayed significantly in the high‐dose group (p < 0.05). Swimming head angle on PND 7 of male offspring was delayed significantly in a dose‐related manner (p < 0.05). Spontaneous behavior examination in males indicated that rearing increased in the high‐dose group in the F₁ generation. The dose levels of PBO in the present study produced some adverse effects in neurobehavioral parameters in mice     

Effects of Maternal Exposure to Imazalil on Behavioral Development in F1‐Generation Mice

Female mice were exposed maternally to imazalil through diet to provide levels of 0 (control), 0.0006, 0.0018, and 0.0054% during gestation and lactation periods, and selected reproductive and neurobehavioral parameters were measured in F₁ generation. There was no adverse effect of imazalil on litter size, litter weight, or sex ratio at birth. With respect to behavioral developmental parameters, surface righting on postnatal day 4 of male offspring was delayed significantly in a dose‐related manner (p < 0.05). Regarding exploratory behavior in the F₁ generation, movement time was significantly long (p = 0.0206) in the low‐dose group of males at 8 weeks of age. Spontaneous behavior examination in males indicated that movement time increased but in females decreased in the low‐dose groups in the F₁ generation. The dose levels of imazalil in the present study produced some adverse effects in neurobehavioral parameters in mice.     

Reproductive and Neurobehavioral Effects of Clothianidin Administered to Mice in the Diet

Clothianidin was given in the diet to provide levels of 0% (control), 0.003%, 0.006%, and 0.012% from 5 weeks of age of the F₀ generation to 11 weeks of age of the F₁ generation in mice. Selected reproductive and neurobehavioral parameters were measured. In exploratory behavior in the F₀ generation, average time of movement, number of rearing, and rearing time of adult males increased significantly in a dose‐related manner. There was no adverse effect of clothianidin on litter size, litter weight, or sex ratio at birth. The average body weight of male and female offspring was increased significantly in a dose‐related manner during the early lactation period. With respect to behavioral developmental parameters, swimming head angle at postnatal day (PND) 7 of male offspring was accelerated significantly in a dose‐related manner. Negative geotaxis at PND 7 of female offspring was accelerated significantly in a dose‐related manner. For movement activity of exploratory behavior in the F₁ generation, number of rearing of female offspring increased significantly in a dose‐related manner. Movement time of adult males increased significantly in a dose‐related manner. The dose levels of clothianidin in the present study produced several adverse effects in neurobehavioral parameters in mice. Nevertheless, it would appear that the levels of the actual dietary intake of clothianidin are unlikely to produce adverse effects in humans.     

Reproductive and Neurobehavioral Effects of Brilliant Blue FCF in Mice

Brilliant blue FCF of food color was given in the diets of mice at levels of 0% (control), 0.08, 0.24, and 0.72% from 5 weeks of age in the F₀ generation and continuing to 11 weeks of age in the F₁ generation and selected reproductive and neurobehavioral parameters were measured. Mice were mated at 9 weeks of age and dams were delivered offspring at 12 weeks of age. Offspring were weaned at 4 weeks of age. Regarding exploratory behavior at 8 weeks of age in the F₀ generation, movement time (sec) displayed a significant tendency to be increased and the average time of rearing (sec) displayed a significant tendency to be decreased in females in the treatment groups in a trend test (p = 0.019 and 0.027, respectively). In the F₁ generation, the development of surface righting at postnatal day 4 was delayed significantly in the high‐dose group (0.72%) in male and female offspring, and those effects were significantly related to dose in a trend test (p< 0.01 for both). Regarding exploratory behavior at 8 weeks of age in the F₁ generation, the number of horizontal activities exhibited a significant tendency to be decreased in females in the treatment groups in a trend test (p = 0.015). Regarding spontaneous behavior, average time of movement (sec) was significantly accelerated in females in the high‐dose group. The dose levels of brilliant blue FCF used in the present study produced a few significant effects on neurobehavioral parameters in multiple generations in mice.     

Effects of Different Types of Bedding Materials on Behavioral Development in Laboratory CD1 Mice (Mus musculus)

Male and female mice were housed in cages, containing different types of bedding materials (wood flakes or pulp chips), from 4 weeks of age in the F₀ generation to 11 weeks of age in the F₁ generation; selected reproductive and neurobehavioral parameters were measured in the F₁ generation. There were no adverse effects of bedding materials on litter size, litter weight, or sex ratios at the time of birth. With regard to behavioral development parameters, bedding materials did not influence any variables (p > 0.05) in both sexes. Regarding exploratory behavior in the F₁ generation, number of defecations significantly varied (p = 0.0203) with bedding materials in males at 3 weeks of age. The number of horizontal activities also significantly varied (p = 0.0342) with bedding materials in males at 8 weeks of age. Multiple‐T water maze performance data indicated that the time required was significantly shortened across trials in pulp chips group than wood flakes group in males (p = 0.0211). Moreover, all spontaneous behavior variables in males significantly varied with bedding materials, particularly the average time of movement was significantly different (p = 0.0037) in distance between parallel lines of types of bedding materials in the F₁ generation. The present study shows that bedding materials influence the neurobehavioral development in mice     

Resistance to change and vulnerability to stress: autistic‐like features of GAP43‐deficient mice

There is an urgent need for animal models of autism spectrum disorder (ASD) to understand the underlying pathology and facilitate development and testing of new treatments. The synaptic growth‐associated protein‐43 (GAP43) has recently been identified as an autism candidate gene of interest. Our previous studies show many brain abnormalities in mice lacking one allele for GAP43 [GAP43 (+/?)] that are consistent with the disordered connectivity theory of ASD. Thus, we hypothesized that GAP43 (+/?) mice would show at least some autistic‐like behaviors. We found that GAP43 (+/?) mice, relative to wild‐type (+/+) littermates, displayed resistance to change, consistent with one of the diagnostic criteria for ASD. GAP43 (+/?) mice also displayed stress‐induced behavioral withdrawal and anxiety, as seen in many autistic individuals. In addition, both GAP43 (+/?) mice and (+/+) littermates showed low social approach and lack of preference for social novelty, consistent with another diagnostic criterion for ASD. This low sociability is likely because of the mixed C57BL/6J 129S3/SvImJ background. We conclude that GAP43 deficiency leads to the development of a subset of autistic‐like behaviors. As these behaviors occur in a mouse that displays disordered connectivity, we propose that future anatomical and functional studies in this mouse may help uncover underlying mechanisms for these specific behaviors. Strain‐specific low sociability may be advantageous in these studies, creating a more autistic‐like environment for study of the GAP43‐mediated deficits of resistance to change and vulnerability to stress.     

The Potential of Cr3 [Triaqua‐μ3‐Oxo‐Hexa‐μ‐Propionatotrichromium(III) Chloride] to Reduce Birth Defects in the Offspring of Diabetic CD‐1 Mice

Diabetes mellitus is a growing concern worldwide and leads to multiple complications during pregnancy. Pharmacologic doses of chromium (Cr) have been linked with improving insulin sensitivity and other positive benefits in the treatment of diabetes in animal models. By using streptozotocin induced hyperglycemia in female CD‐1 mice, reproductive outcomes of diabetic and chromium‐dosed diabetic females were examined. After dosing 10 mg/kg Cr in the form of triaqua‐μ₃‐oxo‐hexa‐μ‐propionatotrichromium(III) chloride or Cr3 during gestation days 8–16 (GD8–GD16), all females were sacrificed on gestation day 17 (GD17) and examined for maternal weight gain. The fetuses were examined for gross malformations and for skeletal malformations. The offspring of Cr3‐dosed females tended to have a reduction in the incidence of supernumerary ribs. While hyperglycemia still had negative impacts on the health of dams and their offspring, administration of Cr led to an apparent trend in the reduction in the number of malformations and incidence of supernumerary ribs compared to those of untreated diabetic mothers     

Effects of prenatal exposure to a 50‐Hz magnetic field on one‐trial passive avoidance learning in 1‐day‐old chicks

We investigated memory impairment in newly hatched chicks following in ovo exposure to a 50‐Hz magnetic field (MF) of 2 mT (60 min/day) on embryonic days 12–18. Isolated and paired chicks were used to test the effect of stress during training, and memory retention was tested at 10, 30, and 120 min, following exposure to a bitter‐tasting bead (100% methylanthranilate). Results showed that memory was intact at 10 min in both isolated and paired chicks with or without MF exposure. However, while isolated chicks had good memory retention levels at 30 and 120 min, those exposed to MF did not. The results suggest a potential disruption of memory formation following in ovo exposure to MF, with this effect only evident in the more stressed, isolated chicks. Bioelectromagnetics 31:150–155, 2010. © 2009 Wiley‐Liss, Inc.     

The effects of 50 Hz magnetic field exposure on dimethylbenz(α)anthracene induced thymic lymphoma/leukemia in mice

Several epidemiological investigations have suggested an increased incidence of lymphoma, leukemia, and brain tumor in residents living near power transmission lines. However, some observers failed to confirm such a positive correlation. To evaluate the effects of extremely low frequency (ELF) magnetic fields on leukemogenesis, an experimental animal model was used, in which thymic lymphoma/leukemia was induced by dimethylbenz(α)anthracene (DMBA) injected subcutaneously into the interscapular region of newborn mice within 24 h after birth. Beginning at the second week of age, 165 mice were exposed to 50 Hz magnetic field at 1 mT, 3 h/day, 6 days/week for 16 weeks, and 155 animals exposed to sham conditions. All surviving animals were killed by cervical dislocation at the age of 32 weeks and were examined pathologically. The results showed that the incidences of advanced thymic lymphoma, complicated with lymphomatous leukemia, were 21.8 and 23.9% in the two groups, respectively, without statistically significant differences. But dense metastatic infiltration by lymphoma cells into liver in the field exposure group greater (50%) than that in the sham-exposure group (16.2%) was observed (χ² = 9.847, P < 0.01). To determine whether ELF acts as a tumor promoter, further experiments are required. Bioelectromagnetics 18:360–364, 1997. © 1997 Wiley-Liss, Inc.     

短时高温对烟蚜生长发育、繁殖和取食行为的影响

【目的】烟蚜Myzus persicae是一种世界性的重要农业害虫,本研究测定了短时高温对烟蚜生长发育、繁殖和取食行为的影响。【方法】测定初产若蚜每天在30, 35和40℃下处理1, 2和4 h的发育历期及成蚜在相同处理下的繁殖力,应用刺吸电位技术（electronic penetration graph, EPG）测定成蚜在30和35℃下处理1和2 h后以及在25, 28, 30和35℃恒温下的取食行为。【结果】短时高温（30~35℃）对各龄若蚜发育历期影响较小,但40℃下处理2 h后其发育历期显著增加（P<0.05）;若蚜存活率、成蚜寿命及其繁殖历期随温度和处理时间的增加而降低;在25~35℃下,随温度和处理时间的增加,成蚜繁殖率变化较小,但40℃下处理2和4 h后,其繁殖率显著降低（P< 0.01）;短时高温对烟蚜的取食行为影响显著,随温度和处理时间的增加,非刺探波np数量（P<0.05）及其持续时间（P<0.01）显著减少,而韧皮部刺吸波E持续时间显著增加（P<0.05）;但持续高温下,随温度升高,np波数量及其持续时间先减少后增加,而E波则相反。【结论】短时高温对烟蚜若蚜发育速率影响较小,但使若蚜存活率、成蚜寿命及其繁殖历期显著降低,且对成蚜繁殖有一定的抑制作用,而对成蚜的取食有一定的促进作用。研究结果有助于了解烟蚜不同年份间夏季种群数量的变化机制。     

PA28α overexpressing female mice maintain exploratory behavior and capacity to prevent protein aggregation in hippocampus as they age

With age, protein damage accumulates and increases the risk of age‐related diseases. The proteasome activator PA28αβ is involved in protein damage clearance during early embryogenesis and has demonstrated protective effects against proteinopathy. We have recently discovered that adult female mice overexpressing PA28α (PA28αOE) have enhanced learning and memory, and protein extracts from their hippocampi prevent aggregation more efficiently than wild type. In this study, we investigated the effect of overexpressing PA28α on aging using C57BL/6N×BALB/c F2 hybrid mice. We found that the hippocampal anti‐aggregation effect was maintained in young adult (7 months) to middle‐aged (15 months) and old (22 months) PA28αOE females. While the PA28αOE influence on learning and memory gradually decreased with aging, old PA28αOE females did not display the typical drop in explorative behavior—a behavioral hallmark of aging—but were as explorative as young mice. PA28αOE lowered PA28‐dependent proteasome capacity in both heart and hippocampus, and there was no indication of lower protein damage load in PA28αOE. The life span of PA28αOE was also similar to wild type. In both wild type and PA28αOE, PA28‐dependent proteasome capacity increased with aging in the heart, while 26S and 20S proteasome capacities were unchanged in the timepoints analyzed. Thus, PA28αOE females exhibit improved hippocampal ability to prevent aggregation throughout life and enhanced cognitive capabilities with different behavioral outcomes dependent on age; improved memory at early age and a youth‐like exploration at old age. The cognitive effects of PA28αβ combined with its anti‐aggregation molecular effect highlight the therapeutical potential of PA28αβ in combating proteinopathies.     

Circulating levels of α‐tocopherol and retinol in free‐ranging African elephants (Loxodonta africana)

To date, there are no detailed reports of circulating levels of plasma α‐tocopherol and retinol for large samples of free‐ranging African elephants (Loxodonta africana). This survey study measured natural circulating levels of α‐tocopherol as a measure of vitamin E activity and retinol as an indicator of vitamin A activity, in 70 free‐ranging African elephants captured at Kruger National Park as part of a translocation program. Mean levels of α‐tocopherol and retinol were found to be 0.613 ± 0.271 μg/mL and 0.039± 0.007 μg/mL, respectively, and did not vary significantly across sex or age class. Elephants appear to normally have low circulating levels of both these nutrients compared with domestic herbivore species; values from healthy, free‐ranging elephant populations may provide useful data for assessing nutrient status of captive animals. Zoo Biol 18:319–323, 1999.© 1999 Wiley‐Liss, Inc.     

Genetic manipulation of the γ‐aminobutyric acid (GABA) shunt in rice: overexpression of truncated glutamate decarboxylase (GAD2) and knockdown of γ‐aminobutyric acid transaminase (GABA‐T) lead to sustained and high levels of GABA accumulation in rice kernels

Gamma‐aminobutyric acid (GABA) is a non‐protein amino acid commonly present in all organisms. Because cellular levels of GABA in plants are mainly regulated by synthesis (glutamate decarboxylase, GAD) and catabolism (GABA‐transaminase, GABA‐T), we attempted seed‐specific manipulation of the GABA shunt to achieve stable GABA accumulation in rice. A truncated GAD2 sequence, one of five GAD genes, controlled by the glutelin (GluB‐1) or rice embryo globulin promoters (REG) and GABA‐T‐based trigger sequences in RNA interference (RNAi) cassettes controlled by one of these promoters as well, was introduced into rice (cv. Koshihikari) to establish stable transgenic lines under herbicide selection using pyriminobac. T₁ and T₂ generations of rice lines displayed high GABA concentrations (2–100 mg/100 g grain). In analyses of two selected lines from the T₃ generation, there was a strong correlation between GABA level and the expression of truncated GAD2, whereas the inhibitory effect of GABA‐T expression was relatively weak. In these two lines both with two T‐DNA copies, their starch, amylose, and protein levels were slightly lower than non‐transformed cv. Koshihikari. Free amino acid analysis of mature kernels of these lines demonstrated elevated levels of GABA (75–350 mg/100 g polished rice) and also high levels of several amino acids, such as Ala, Ser, and Val. Because these lines of seeds could sustain their GABA content after harvest (up to 6 months), the strategy in this study could lead to the accumulation GABA and for these to be sustained in the edible parts.     

In Vitro and In Vivo Evaluation of N‐{2‐[4‐(3‐Cyanopyridin‐2‐yl)piperazin‐1‐yl]ethyl}‐3‐[11C]methoxybenz­amide,a Positron Emission Tomography (PET) Radioligand for Dopamine D4 Receptors,in Rodents

The D₄ dopamine receptor belongs to the D₂‐like family of dopamine receptors, and its exact regional distribution in the central nervous system is still a matter of considerable debate. The availability of a selective radioligand for the D₄ receptor with suitable properties for positron emission tomography (PET) would help resolve issues of D₄ receptor localization in the brain, and the presumed diurnal change of expressed protein in the eye and pineal gland. We report here on in vitro and in vivo characteristics of the high‐affinity D₄ receptor‐selective ligand N‐{2‐[4‐(3‐cyanopyridin‐2‐yl)piperazin‐1‐yl]ethyl}‐3‐[¹¹C]methoxybenzamide ([¹¹C] 2 ) in rat. The results provide new insights on the in vitro properties that a brain PET dopamine D₄ radioligand should possess in order to have improved in vivo utility in rodents.     

Sensitive biomarker of polycyclic aromatic hydrocarbons (PAHs): urinary 1-hydroxyprene glucuronide in relation to smoking and low ambient levels of exposure

The lysosomal neutral red retention time (NRRT) assay, a biomarker for lysosomal membrane stability, and the total immune activity (TIA) assay, a measure of non-specific immune system activity, were used in laboratory studies to assess the toxic effects of 2,4,6-trinitrotoluene (TNT) on earthworms (Eisenia andrei) in vivo. The results were compared with the concentration of TNT and its metabolites in earthworm tissue, as well as standard sublethal toxicity endpoints including growth (i.e. weight change) and reproduction effects from previously published studies. Filter paper experiments indicated a significant decrease in NRRT at ≥1.8 μg TNT cm^-2, whereas sublethal (weight loss) and lethal effects to earthworms were detected at ≥3.5 and 7.1 μg TNT cm^-2, respectively. Experiments in artificial soil showed that NRRT effects could be detected at lower TNT concentrations (≥55 mg TNT kg^-1 soil dry weight) compared with other sublethal endpoints (effects on growth and reproduction). The TIA biomarker did not significantly respond to TNT. Copper (as CuSO₄, filter paper contact tests) and 2-chloroacetamide (soil tests), which were used as reference toxicants, also decreased the NRRT. The use of the NRRT assay linked with tissue concentrations of TNT metabolites in earthworms was identified as a potentially appropriate biomarker approach for TNT exposure assessment under laboratory conditions and a novel tool for effects-based risk assessment.     

Age‐related deterioration of motor function in male and female 5xFAD mice from 3 to 16 months of age

Alzheimer's disease (AD) is a neurodegenerative disorder that leads to age‐related cognitive and sensori‐motor dysfunction. There is an increased understanding that motor dysfunction contributes to overall AD severity, and a need to ameliorate these impairments. The 5xFAD mouse develops the neuropathology, cognitive and motor impairments observed in AD, and thus may be a valuable animal model to study motor deficits in AD. Therefore, we assessed age‐related changes in motor ability of male and female 5xFAD mice from 3 to 16 months of age, using a battery of behavioral tests. At 9‐10 months, 5xFAD mice showed reduced body weight, reduced rearing in the open‐field and impaired performance on the rotarod compared to wild‐type controls. At 12‐13 months, 5xFAD mice showed reduced locomotor activity on the open‐field, and impaired balance on the balance beam. At 15‐16 months, impairments were also seen in grip strength. Although sex differences were observed at specific ages, the development of motor dysfunction was similar in male and female mice. Given the 5xFAD mouse is commonly on a C57BL/6 × SJL hybrid background, a subset of mice may be homozygous recessive for the Dysf ^im mutant allele, which leads to muscular weakness in SJL mice and may exacerbate motor dysfunction. We found small effects of Dysf ^im on motor function, suggesting that Dysf ^im contributes little to motor dysfunction in 5xFAD mice. We conclude that the 5xFAD mouse may be a useful model to study mechanisms that produce motor dysfunction in AD, and to assess the efficacy of therapeutics on ameliorating motor impairment.     

A validated HPLC method for the determination of dimethyl‐4,4′‐dimethoxy‐5,6,5′,6′‐dimethylene dioxybiphenyl‐2,2′‐dicarboxylate (DDB) with fluorescence detection in raw material and pill form: application to an in vitro dissolution test and a content uniformity test

A simple, sensitive and rapid HPLC method with fluorescence detection for the determination of dimethyl‐4,4′‐dimethoxy‐5,6,5′,6′‐dimethylene dioxybiphenyl‐2,2′‐dicarboxylate (DDB) in the raw material and pill form was developed. Liquid chromatography was performed on a C₁₈ column (250 × 4.6 mm i.d., 5 µm particle size), the mobile phase consisted of methanol and 0.05 M sodium dihydrogen phosphate buffer (80 : 20, v/v), and the apparent pH of the mobile phase was adjusted to 3. The fluorescence detector was operated at excitation/emission wavelengths of 275/400 nm. The proposed method allows the determination of DDB within concentration range 0.1–1.5 µg/mL with a limit of detection of 0.032 µg/mL, a limit of quantification of 0.097 µg/mL and a correlation coefficient of 0.9997. The proposed method has been successfully applied for the analysis of DDB in its pills with a percentage recovery of 98.45 ± 0.32. The method was fully validated according to ICH guidelines. Moreover, the high sensitivity of the method permits its use in an in vitro dissolution test for DDB under simulated intestinal conditions. In addition, the proposed method was extended to a content uniformity test according to USP guidelines. Copyright © 2014 John Wiley & Sons, Ltd.