Injurious Effects of Curcumin on Maturation of Mouse Oocytes, Fertilization and Fetal Development via Apoptosis

Curcumin, a common dietary pigment and spice, is a hydrophobic polyphenol derived from the rhizome of the herb Curcuma longa. Previously, we reported a cytotoxic effect of curcumin on mouse embryonic stem cells and blastocysts and its association with defects in subsequent development. In the present study, we further investigated the effects of curcumin on oocyte maturation and subsequent pre- and post-implantation development, both in vitro and in vivo. Notably, curcumin induced a significant reduction in the rate of oocyte maturation, fertilization, and in vitro embryonic development. Treatment of oocytes with curcumin during in vitro maturation (IVM) led to increased resorption of postimplantation embryos and decreased fetal weight. Experiments with an in vivo mouse model disclosed that consumption of drinking water containing 40 μM curcumin led to decreased oocyte maturation and in vitro fertilization as well as early embryonic developmental injury. Finally, pretreatment with a caspase-3-specific inhibitor effectively prevented curcumin-triggered injury effects, suggesting that embryo impairment by curcumin occurs mainly via a caspase-dependent apoptotic process.     

Injurious Effects of Emodin on Maturation of Mouse Oocytes,Fertilization and Fetal Development via Apoptosis

Emodin (1,3,8-trihydroxy-6-methylanthraquinone), a major constituent of rhubarb, has a wide range of therapeutic applications. Previous studies have established that emodin induces apoptosis in the inner cell mass and trophectoderm of mouse blastocysts and leads to decreased embryonic development and viability, indicating a role as an injury risk factor for normal embryonic development. However, the mechanisms underlying its hazardous effects have yet to be characterized. In the current study, we further investigated the effects of emodin on oocyte maturation and subsequent pre- and post-implantation development, both in vitro and in vivo. Notably, emodin induced a significant reduction in the rates of oocyte maturation, fertilization, and in vitro embryonic development. Treatment of oocytes with emodin during in vitro maturation (IVM) led to increased resorption of postimplantation embryos and decreased fetal weight. Experiments using an in vivo mouse model disclosed that consumption of drinking water containing 20–40 μM emodin led to decreased oocyte maturation and in vitro fertilization, as well as early embryonic developmental injury. Notably, pretreatment with a caspase-3-specific inhibitor effectively prevented emodin-triggered injury effects, suggesting that impairment of embryo development occurs via a caspase-dependent apoptotic process.     

Cytotoxic Effects of CdSe Quantum Dots on Maturation of Mouse Oocytes, Fertilization, and Fetal Development

Quantum dots (QDs) are useful novel luminescent markers, but their embryonic toxicity is yet to be fully established, particularly in oocyte maturation and sperm fertilization. Earlier experiments by our group show that CdSe-core QDs have cytotoxic effects on mouse blastocysts and are associated with defects in subsequent development. Here, we further investigate the influence of CdSe-core QDs on oocyte maturation, fertilization, and subsequent pre- and postimplantation development. CdSe-core QDs induced a significant reduction in the rates of oocyte maturation, fertilization, and in vitro embryo development, but not ZnS-coated CdSe QDs. Treatment of oocytes with 500 nM CdSe-core QDs during in vitro maturation (IVM) led to increased resorption of postimplantation embryos and decreased placental and fetal weights. To our knowledge, this is the first study to report the negative impact of CdSe-core QDs on mouse oocyte development. Moreover, surface modification of CdSe-core QDs with ZnS effectively prevented this cytotoxicity.     

Cytotoxic Effects of Dillapiole on Embryonic Development of Mouse Blastocysts in Vitro and in Vivo

We examined the cytotoxic effects of dillapiole, a phenylpropanoid with antileishmanial, anti-inflammatory, antifungal, and acaricidal activities, on the blastocyst stage of mouse embryos, subsequent embryonic attachment and outgrowth in vitro, and in vivo implantation via embryo transfer. Blastocysts treated with 2.5–10 μM dillapiole exhibited a significant increase in apoptosis and corresponding decrease in total cell number. Notably, the implantation success rates of blastocysts pretreated with dillapiole were lower than those of their control counterparts. Moreover, in vitro treatment with 2.5–10 μM dillapiole was associated with increased resorption of post-implantation embryos and decreased fetal weight. Our results collectively indicate that dillapiole induces apoptosis and retards early post-implantation development, both in vitro and in vivo. However, the extent to which this organic compound exerts teratogenic effects on early human development is not known at present. Further studies are required to establish effective protection strategies against the cytotoxic effects of dillapiole.     

Hazardous Effects of Curcumin on Mouse Embryonic Development through a Mitochondria-Dependent Apoptotic Signaling Pathway

In this study, we examined the cytotoxic effects of curcumin, the yellow pigment of Curcuma longa, on the blastocyst stage of mouse embryos, subsequent embryonic attachment, and outgrowth in vitro and in vivo implantation by embryo transfer. Mouse blastocysts were incubated in medium with or without curcumin (6, 12 or 24 μM) for 24 h. Cell proliferation and growth were investigated using dual differential staining, apoptosis was analyzed with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), and implantation and post-implantation development of embryos were measured by in vitro development analysis and in vivo embryo transfer, respectively. Blastocysts treated with 24 μM curcumin displayed significantly increased apoptosis and decreased total cell number. Interestingly, we observed no marked differences in the implantation success rates between curcumin-pretreated and control blastocysts during in vitro embryonic development through implantation with a fibronectin-coated culture dish. However, in vitro treatment with 24 μM curcumin was associated with decreased implantation rate and increased resorption of postimplantation embryos in mouse uterus, as well as decreased fetal weight in the embryo transfer assay. Our results collectively indicate that in vitro exposure to curcumin triggers apoptosis and retards early postimplantation development after transfer to host mice. In addition, curcumin induces apoptotic injury effects on mouse blastocysts through ROS generation, and further promotes mitochondria-dependent apoptotic signaling processes to impair sequent embryonic development.     

Antiproliferative and apoptotic effects of tocopherols and tocotrienols on normal mouse mammary epithelial cells

Studies were conducted to determine the comparative effects of tocopherols and tocotrienols on normal mammary epithelial cell growth and viability. Cells isolated from midpregnant BALB/c mice were grown within collagen gels and maintained on serum-free media. Treatment with 0–120 μM α-and γ-tocopherol had no effect, whereas 12.5–100 m μM tocotrienol-rich fraction of palm oil (TRF), 100–120 μM δ-tocopherol, 50–60 μM α-tocotrienol, and 8–14 μM γ- or δ-tocotrienol significantly inhibited cell growth in a dose-responsive manner. In acute studies, 24-h exposure to 0–250 μM α-, γ-, and δ-tocopherol had no effect, whereas similar treatment with 100–250 μM TRF, 140–250 μM α-, 25–100 μM γ- or δ-tocotrienol significantly reduced cell viability. Growth-inhibitory doses of TRF, δ-tocopherol, and a-, γ-, and δ-tocotrienol were shown to induce apoptosis in these cells, as indicated by DNA fragmentation. Results also showed that mammary epithelial cells more easily or preferentially took up tocotrienols as compared to tocopherols, suggesting that at least part of the reason tocotrienols display greater biopotency than tocopherols is because of greater cellular accumulation. In summary, these findings suggest that the highly biopotent γ- and δ-tocotrienol isoforms may play a physiological role in modulating normal mammary gland growth, function, and remodeling.     

Potential Development of Vitrified Immature Human Oocytes: Influence of the Culture Medium and the Timing of Vitrification

How does the in vitro maturation (IVM) medium and the vitrification procedure affect the survival of germinal vesicle (GV) oocytes obtained from stimulated cycles and their development to the blastocyst stage? In total, 1085 GV human oocytes were obtained after women underwent a cycle of controlled ovarian stimulation, and these oocytes were subjected to IVM before or after their vitrification. IVM was carried out in two commercial culture media not specifically designed for maturation. MII oocytes were then activated and embryo development until day 6 was evaluated. According to the results, a higher percentage of oocytes reach the MII stage if they are vitrified before they undergo IVM. Nevertheless, the medium used and the sample size determine whether these differences become significant or not. Similar survival rates and development to blastocysts were observed in all the conditions studied.     

二氧化锡薄膜的制备和应用研究进展

介绍了近年来二氧化锡薄膜的制备方法和在功能材料中的应用。探讨了各种方法的特点，提出了今后二氧化锡薄膜的发展趋势和应用前景。     

Resveratrol Protects against 2-Bromopropane-Induced Apoptosis and Disruption of Embryonic Development in Blastocysts

2-Bromopropane (2-BP) is used as an alternative to ozone-depleting cleaning solvents. Previously, we reported that 2-BP has cytotoxic effects on mouse blastocysts and is associated with defects in subsequent development. In the present work, we show that 2-BP induces apoptosis in the inner cell mass of mouse blastocysts, and inhibits cell proliferation. Both effects are suppressed by resveratrol, a grape-derived phytoalexin with known antioxidant and anti-inflammatory properties. 2-BP-treated blastocysts displayed lower levels of implantation (compared to controls) when plated on culture dishes in vitro, and a reduced ability to proceed to later stages of embryonic development. Pretreatment with resveratrol prevented 2-BP-induced disruption of embryonic development, both in vitro and in vivo. Further investigation of these processes revealed that 2-BP directly promotes ROS generation, loss of mitochondrial membrane potential (MMP), and activation of caspase-3, whereas resveratrol effectively blocks 2-BP-induced ROS production and the accompanying apoptotic biochemical changes. Our results collectively imply that 2-BP triggers the mitochondrion-dependent apoptotic pathway via ROS generation, and the antioxidant activity of resveratrol prevents 2-BP-induced toxicity.     

Effect of natural coagula maturation on the processability,cure, and mechanical properties of unfilled vulcanizates of Hevea natural rubber

Fresh latex from different Hevea brasiliensis clones was naturally coagulated, subjected to different durations of maturation, processed into solid rubber, and compounded into pure gum stocks and vulcanized. Coagula maturation had clone‐specific effects on the processability of the raw rubber: reduced for some clones, while others was less sensitive. The cure and mechanical behaviors of the compounded stocks and vulcanizates, respectively, were not sensitive to the clonal origin of coagula and their duration of maturation. Although coagula maturation could be associated with leaching, deactivation of inherent antioxidants in Hevea latex, as well as crosslinking and/or oxidation of polyisoprene chains, these results show that compounding with a standard pure gum recipe compensates for the Hevea latex constituents affected by maturation. Hence, extended maturation of Hevea coagula, for economic or other reasons, would influence much more the bulk behavior of raw rubber and have insignificant effects on the compounded stocks and vulcanizates. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 2359–2363, 2007     

The Role of MAPK3/1 and AKT in the Acquisition of High Meiotic and Developmental Competence of Porcine Oocytes Cultured In Vitro in FLI Medium

The developmental potential of porcine oocytes cultured in vitro was remarkably enhanced in a medium containing FGF2, LIF and IGF1 (FLI) when compared to a medium supplemented with gonadotropins and EGF (control). We analyzed the molecular background of the enhanced oocyte quality by comparing the time course of MAPK3/1 and AKT activation, and the expression of genes controlled by these kinases in cumulus-oocyte complexes (COCs) cultured in FLI and the control medium. The pattern of MAPK3/1 activation in COCs was very similar in both media, except for a robust increase in MAPK3/1 phosphorylation during the first hour of culture in the FLI medium. The COCs cultured in the FLI medium exhibited significantly higher activity of AKT than in the control medium from the beginning up to 16 h of culture; afterwards a deregulation of AKT activity occurred in the FLI medium, which was not observed in the control medium. The expression of cumulus cell genes controlled by both kinases was also modulated in the FLI medium, and in particular the genes related to cumulus-expansion, signaling, apoptosis, antioxidants, cell-to-cell communication, proliferation, and translation were significantly overexpressed. Collectively, these data indicate that both MAPK3/1 and AKT are implicated in the enhanced quality of oocytes cultured in FLI medium.     

Early Expression of Tet1 and Tet2 in Mouse Zygotes Altered DNA Methylation Status and Affected Embryonic Development

Ten-eleven translocation (Tet) dioxygenases can induce DNA demethylation by catalyzing 5-methylcytosine(5mC) to 5-hydroxymethylcytosine(5hmC), and play important roles during mammalian development. In mouse, Tet1 and Tet2 are not expressed in pronucleus-staged embryos and are not involved in the genomic demethylation of early zygotes. Here, we investigated the influence of Tet1 and Tet2 on methylation of parental genomes by ectopically expressing Tet1 and Tet2 in zygotes. Immunofluorescence staining showed a marked 5hmC increase in the maternal pronucleus after injection of Tet1 or Tet2 mRNA into zygotes. Whole-genome bisulfite sequencing further revealed that Tet2 greatly enhanced the global demethylation of both parental genomes, while Tet1 only promoted the paternal demethylation. Tet1 and Tet2 overexpression altered the DNA methylation across genomes, including various genic elements and germline-specific differently methylated regions. Tet2 exhibited overall stronger demethylation activity than Tet1. Either Tet1 or Tet2 overexpression impaired preimplantation embryonic development. These results demonstrated that early expression of Tet1 and Tet2 could substantially alter the zygotic methylation landscape and damage embryonic development. These findings provide new insights into understanding the function of Tet dioxygenases and the mechanism of DNA methylation in relation to embryogenesis.     

乙型肝炎病毒X蛋白对小鼠胚胎肝干细胞凋亡及相关蛋白表达的影响

目的研究乙型肝炎病毒(hepatitis B virus,HBV)X蛋白(HBx)对小鼠胚胎肝干细胞(embryonic liver stemcell,ELSC)凋亡及相关蛋白Bcl2、Mcl1、Bax表达的影响。方法采用表达绿色荧光蛋白(green fluorescent protein,GFP)的腺病毒载体系统将HBx基因转入小鼠胚胎肝干细胞ELSC14.5中,采用RT-PCR和Western blot法检测细胞中HBx基因mRNA的转录和蛋白的表达;Hoechst33342染色法观察细胞核的改变;TUNEL法和流式细胞术检测细胞的凋亡情况;Real-time PCR和Western blot法检测抗凋亡因子Bcl2、Mcl1和促凋亡因子Bax基因mRNA的转录水平和蛋白的表达水平。结果重组腺病毒Ad-GFP-HBx能有效感染ELSC14.5细胞,HBx基因和蛋白均能特异性表达;感染的ELSC14.5细胞核呈现固缩,且边缘化的细胞数减少,细胞凋亡率降低;细胞中Bcl2和Mcl1基因mRNA的转录水平和蛋白的表达水平均增高,而Bax的表达降低。结论 HBx可通过调节Bcl2家族中抗凋亡因子和促凋亡因子的比例失衡,来抑制小鼠胚胎肝干细胞的凋亡,促进其存活。     

Penicillium bilaji and phosphorus fertilization effects on the growth,development, yield and common root rot severity of spring wheat

A strain ofPenicillium bilaji Chalabuda (PB) has recently been commercially released as a seed inoculant to increase phosphorus (P) uptake by wheat (Triticum aestivum L.). The purpose of this study was to compare the effects of drill applied P (15 kg P ha^–1) with PB seed inoculation on early growth, development, P uptake, and grain yield of Stoa spring wheat at four sites in North Dakota.Fertilization with P consistently enhanced early season growth, main stem development, tillering and P uptake. Seed inoculation with PB had little or no effect on these traits. Phosphorus fertilization tended to increase common root rot severity (CRR, incited byCochliobolus sativus (Ito & Kurib) Drechs.), while PB inoculation had no effect. Grain yields were significantly increased by P fertilization at one location. Inoculation with PB also increased grain yield at this location. The reason why PB inoculation increased yield at this location is not evident, as plant growth and P uptake were not enhanced earlier in the season. Averaged across all four sites, PB inoculation increased wheat yields 66 kg ha^–1, which is similar to averaged yield responses reported from the Prairie Provinces of Canada.     

Multiparametric analysis of anti-proliferative and apoptotic effects of gold nanoprisms on mouse and human primary and transformed cells,biodistribution and toxicity in vivo

Background

The special physicochemical properties of gold nanoprisms make them very useful for biomedical applications including biosensing and cancer therapy. However, it is not clear how gold nanoprisms may affect cellular physiology including viability and other critical functions. We report a multiparametric investigation on the impact of gold-nanoprisms on mice and human, transformed and primary cells as well as tissue distribution and toxicity in vivo after parental injection.

Methods

Cellular uptake of the gold-nanoprisms (NPRs) and the most crucial parameters of cell fitness such as generation of reactive oxygen species (ROS), mitochondria membrane potential, cell morphology and apoptosis were systematically assayed in cells. Organ distribution and toxicity including inflammatory response were analysed in vivo in mice at 3 days or 4 months after parental administration.

Results

Internalized gold-nanoprisms have a significant impact in cell morphology, mitochondrial function and ROS production, which however do not affect the potential of cells to proliferate and form colonies. In vivo NPRs were only detected in spleen and liver at 3 days and 4 months after administration, which correlated with some changes in tissue architecture. However, the main serum biochemical markers of organ damage and inflammation (TNFα and IFNγ) remained unaltered even after 4 months. In addition, animals did not show any macroscopic sign of toxicity and remained healthy during all the study period.

Conclusion

Our data indicate that these gold-nanoprisms are neither cytotoxic nor cytostatic in transformed and primary cells, and suggest that extensive parameters should be analysed in different cell types to draw useful conclusions on nanomaterials safety. Moreover, although there is a tendency for the NPRs to accumulate in liver and spleen, there is no observable negative impact on animal health.

     

2-氯吡啶和2,6-二氯吡啶合成与应用

介绍了 2 氯吡啶和 2 ,6 氯吡啶的合成技术 ,应用 ,市场与发展     

纳米锌铁氧体研究现状及发展前景

简要介绍了纳米锌铁氧体（ZnFe2O4）的研究现状。详细分析了目前国内外纳米锌铁氧体的主要制备方法及研究进展,比较了各种制备方法的优缺点。结合相关行业的发展,指出纳米锌铁氧体今后的研究方向及发展前景。     

2-氯吡啶和2,6-二氯吡啶

2-氯吡啶和2,6-二氯吡啶是重要的医药、农药、日化用品和染料等的原料,文章 介绍了2-氯吡啶和2,6-二氯吡啶的合成技术,应用,市场与发展。