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.     

Sperm metabolism is altered during storage by female insects: evidence from two-photon autofluorescence lifetime measurements in bedbugs

We explore the possibility of characterizing sperm cells without the need to stain them using spectral and fluorescence lifetime analyses after multi-photon excitation in an insect model. The autofluorescence emission spectrum of sperm of the common bedbug, Cimex lectularius, was consistent with the presence of flavins and NAD(P)H. The mean fluorescence lifetimes showed smaller variation in sperm extracted from the male (tau m, τ_m = 1.54–1.84 ns) than in that extracted from the female sperm storage organ (tau m, τ_m = 1.26–2.00 ns). The fluorescence lifetime histograms revealed four peaks. These peaks (0.18, 0.92, 2.50 and 3.80 ns) suggest the presence of NAD(P)H and flavins and show that sperm metabolism can be characterized using fluorescence lifetime imaging. The difference in fluorescence lifetime variation between the sexes is consistent with the notion that female animals alter the metabolism of sperm cells during storage. It is not consistent, however, with the idea that sperm metabolism represents a sexually selected character that provides females with information about the male genotype.     

Cyclamate intake and cyclohexylamine excretion are not related to male fertility in humans

Cyclamate and its metabolite cyclohexylamine affect male fertility in high dose animal studies, but this affect has not been investigated in epidemiological studies. This paper reports the first epidemiological study designed to investigate the possibility of a relationship between cyclamate and cyclohexylamine and male fertility in humans, in which 405 cases of clinically defined infertility in men and 379 controls were surveyed. Semen evaluation, urine analysis for cyclamate and cyclohexylamine and dietary questionnaires were compared between cases and controls. No evidence was found of a significant association between cyclamate intake and male infertility; neither high cyclamate nor high cyclohexylamine excretion were associated with elevated risk. The lack of association remained after adjusting by age, area of residence, education, total energy intake and other variables. No significant correlations were observed between cyclamate intake, metabolism or excretion, and sperm count and motility. The results demonstrate no effect of cyclamate or cyclohexylamine on male fertility at the present levels of cyclamate consumption.     

Absence makes the adaptations grow fonder: Proportion of time apart from partner, male sexual psychology, and sperm competition in humans (Homo sapiens).

Sperm competition occurs when the sperm of multiple males concurrently occupy the reproductive tract of a female and compete to fertilize an egg. We used a questionnaire to investigate psychological responses to the risk of sperm competition for 237 men in committed, sexual relationships. As predicted, a man who spends a greater (relative to a man who spends a lesser) proportion of time apart from his partner since the couple's last copulation reported (a) greater sexual interest in his partner, (b) greater distress in response to his partner's sexual rejection, and (c) greater sexual persistence in response to his partner's sexual rejection. All effects were independent of total time since the couple's last copulation and the man's relationship satisfaction. Discussion addresses limitations of the current research and situates the current results within the broader comparative literature on adaptation to sperm competition. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

激光辐照对冷冻精子活率和活力的影响

分别用7.48J/cm~2和14.96J/cm~2能量密度的氦氖激光辐照23份液氮冷冻前人的精液,冻贮三个月后复温查精子活率和毛细管穿透高度。结果:经7.48J/cm~2剂量的激光辐照后的精子活率和穿透高度明显提高,分别平均提高10.18％和8.32mm,与未受辐照组比较差异显著(P<0.05)经14.96J/cm~2辐照组,活率较未照组降低。实验结果说明适当剂量的激光辐照能够增强精子对冷冻的耐受力,提高精子的复活率和活动力。     

Telomere Length in Metaphase Chromosomes of Human Triploid Zygotes

The human lifespan is strongly influenced by telomere length (TL) which is defined in a zygote—when two highly specialised haploid cells form a new diploid organism. Although TL is a variable parameter, it fluctuates in a limited range. We aimed to establish the determining factors of TL in chromosomes of maternal and paternal origin in human triploid zygotes. Using Q-FISH, we examined TL in the metaphase chromosomes of 28 human triploid zygotes obtained from 22 couples. The chromosomes’ parental origin was identified immunocytochemically through weak DNA methylation and strong hydroxymethylation in the sperm-derived (paternal) chromosomes versus strong DNA methylation and weak hydroxymethylation in the oocyte-derived (maternal) ones. In 24 zygotes, one maternal and two paternal chromosome sets were identified, while the four remaining zygotes contained one paternal and two maternal sets. For each zygote, we compared mean relative TLs between parental chromosomes, identifying a significant difference in favour of the paternal chromosomes, which attests to a certain “imprinting” of these regions. Mean relative TLs in paternal or maternal chromosomes did not correlate with the respective parent’s age. Similarly, no correlation was observed between the mean relative TL and sperm quality parameters: concentration, progressive motility and normal morphology. Based on the comparison of TLs in chromosomes inherited from a single individual’s gametes with those in chromosomes inherited from different individuals’ gametes, we compared intraindividual (intercellular) and interindividual variability, obtaining significance in favour of the latter and thus validating the role of heredity in determining TL in zygotes. A comparison of the interchromatid TL differences across the chromosomes from sets of different parental origin with those from PHA-stimulated lymphocytes showed an absence of a significant difference between the maternal and paternal sets but a significant excess over the lymphocytes. Therefore, interchromatid TL differences are more pronounced in zygotes than in lymphocytes. To summarise, TL in human zygotes is determined both by heredity and parental origin; the input of other factors is possible within the individual’s reaction norm.     

Uric Acid: The Sperm-Release Pheromone of the Marine Polychaete Platynereis dumerilii

In the marine polychaete, Platynereis dumerilii, reproductive behavior in the two sexes is synchronized by the consecutive discharge of male and female sex-specific pheromones. After the female releases the eggs into the free water column, immediate fertilization is achieved by several males circling around the eggs emitting sperm clouds. We report the isolation and identification of the sperm-release pheromone present in the coelomic fluid of sexually mature females. Each step in isolation was guided by bioassay. Isolation methods included extraction and solvent partitioning and separation methods included ultrafiltration and high-performance liquid chromatography. Uric acid was identified as the sperm-release pheromone that is discharged by the female with release of the eggs. The threshold concentration for sperm release by males was determined as 0.6 M.     

共振散射光谱法研究槲皮素与DNA相互作用

在BR缓冲液中,槲皮素存在微弱的共振散射现象,分别在288 nm、466 nm、562 nm处出现共振散射峰,随着DNA的加入,散射信号逐渐增强,且以上3处的峰值明显增加,并且562 nm处的增加值与DNA在3.024×10-7 mol/L～9.072×10-5 mol/L范围内呈现出良好的线性关系.在20℃时,通过Scatchard方程计算出槲皮素与DNA结合常数K=9.91×106 mol-1.L.     

Ketamine effects on the urogenital system--changes in the urinary bladder and sperm motility

Different doses of ketamine (10 mg/kg, 20 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, and 60 mg/kg) were injected i.p. (I.P.), respectively, to male ICR mice to determine the optimal dosage for chronic administration. At and above 40 mg/kg I.P. injection, mice had almost no hindlimb movement during swimming test. Subsequently, 30 mg/kg was used as the dose for the study in the toxicity of long-term ketamine administration on urinary bladder and sperm motility. The treatment group were subdivided into two (n = 10 each group); one received daily ketamine treatment i.p. for 3 months and another group for 6 months. Corresponding number of mice in control groups (n = 5 each group) received saline injection instead of ketamine. Terminal dUTP nick and labeling (TUNEL) study and Sirius red staining were carried out on the sectioned slides of the urinary bladders to study the degree of apoptosis in both epithelium and muscular layers of the urinary bladder and the relative thickness of the muscular layers in this organ was also computed. Apoptosis in the bladder epithelium was observed initially in the 3-month ketamine treated mice and the number of apoptotic cells was significantly different (P < 0.05) between the 3-month and 6-month ketamine treated mice and the control. The relative thickness of muscular layers in the bladder wall also decreased significantly (P < 0.05) when the 6-month treated mice and the control were compared. Sirius red staining revealed increase of collagen in the urinary bladder of the treated mice, most evidently 6 months after ketamine treatment. In addition, the sperm motility was studied and there was a statistically significant difference between the control and ketamine treated groups in the percentages of sperms which were motile (P < 0.05). This suggested that the chronic administration of ketamine affected the genital system as well.     

白蚁提取液急性毒性及致突变性研究

本文采用大,小鼠急性经口毒性试验、Ａｍｅｓ试验、小鼠骨髓细胞微核试验及小鼠精子畸变试验对白蚁提取液的毒理学安全性进行了研究,结果表明,白蚁提取液大,小鼠急性经口ＬＤ５０均大于１０ｇ／ｋｇ体重,属“实际无毒”：体内（包括体细胞和生殖细胞）、体外致突变性试验系统均未检出其致突性。