Transgenerational Effects of Di(2-Ethylhexyl) Phthalate on Anogenital Distance, Sperm Functions and DNA Methylation in Rat Offspring.

Di(2-ethylhexyl) phthalate (DEHP) is widely used as a plasticizer in the manufacture of polyvinylchloride plastics and has been associated with concerns regarding male reproductive toxicity. In this study, we hypothesized that maternal exposure to DEHP induces transgenerational inheritance of adult-onset adverse reproductive outcomes through the male germline in the F1, F2, and F3 generations of male offspring. Pregnant rats were treated with 5 or 500 mg of DEHP/kg/day through gavage from gestation day 0 to birth. The offspring body weight, anogenital distance (AGD), anogenital index (AGI), sperm count, motility, and DNA fragmentation index (DFI) were measured for all generations. Methyl-CpG binding domain sequencing was performed to analyze sperm DNA methylation status in the F3. DEHP exposure at 500 mg/kg affected AGD, AGI, sperm count, mean DFI, and %DFI in the F1; AGD, sperm count, and mean DFI in the F2; and AGD, AGI, mean DFI, and %DFI in the F3. DEHP exposure at 5 mg/kg affected AGD, AGI, sperm count, and %DFI in the F1; sperm count in the F2; and AGD and AGI in F3. Compared with the control group, 15 and 45 differentially hypermethylated genes were identified in the groups administered 5 mg/kg and 500 mg/kg DEHP, respectively. Moreover, 130 and 6 differentially hypomethylated genes were observed in the groups administered 5 mg/kg and 500 mg/kg DEHP. Overall, these results demonstrated that prenatal exposure to DEHP caused transgenerational epigenetic effects, which may explain the observed phenotypic changes in the male reproductive system.

Microfluidic emulsification and sorting assisted preparation of monodisperse chitosan microparticles.

A microfluidic device for generating monodisperse chitosan microparticles and separating the desired particle from smaller particles created as a byproduct of this process was described. The purpose of this study is to separate the satellite droplets from their parent droplets to enhance the size uniformity of the desired microparticles. A double T-junction design was first employed to control the emulsification and the separation, respectively. The results show that the size and gap of the parent droplets are tunable by adjusting the water and oil flow rates. A separation ratio of the satellite droplets of more than 99% was observed. The proposed microfluidic chip is capable of generating relatively uniform micro-droplets with well controllable diameter, and it has the added advantages of being a simple, low cost, and high throughput process. In the future this apparatus can be used to fabricate size-controlled monodisperse microparticles to act as drug carriers for biotechnology and biomedicine applications.