Cumin Prevents 17β-Estradiol-Associated Breast Cancer in ACI Rats

Breast cancer (BC) is a leading cause of cancer deaths in women in less developed countries and the second leading cause of cancer death in women in the U.S. In this study, we report the inhibition of E2-mediated mammary tumorigenesis by Cuminum cyminum (cumin) administered via the diet as cumin powder, as well as dried ethanolic extract. Groups of female ACI rats were given either an AIN-93M diet or a diet supplemented with cumin powder (5% and 7.5%, w/w) or dried ethanolic cumin extract (1%, w/w), and then challenged with subcutaneous E2 silastic implants (1.2 cm; 9 mg). The first appearance of a palpable mammary tumor was significantly delayed by both the cumin powder and extract. At the end of the study, the tumor incidence was 96% in the control group, whereas only 55% and 45% animals had palpable tumors in the cumin powder and extract groups, respectively. Significant reductions in tumor volume (660 ± 122 vs. 138 ± 49 and 75 ± 46 mm³) and tumor multiplicity (4.21 ± 0.43 vs. 1.16 ± 0.26 and 0.9 ± 0.29 tumors/animal) were also observed by the cumin powder and cumin extract groups, respectively. The cumin powder diet intervention dose- and time-dependently offset E2-related pituitary growth, and reduced the levels of circulating prolactin and the levels of PCNA in the mammary tissues. Mechanistically, the cumin powder diet resulted in a significant reversal of E2-associated modulation in ERα, CYP1A1 and CYP1B1. Further, the cumin powder diet reversed the expression levels of miRNAs (miR-182, miR-375, miR-127 and miR-206) that were highly modulated by E2 treatment. We analyzed the composition of the extract by GC/MS and established cymene and cuminaldehyde as major components, and further detected no signs of gross or systemic toxicity. Thus, cumin bioactives can significantly delay and prevent E2-mediated mammary tumorigenesis in a safe and effective manner, and warrant continued efforts to develop these clinically translatable spice bioactives as chemopreventives and therapeutics against BC.     

紫草多糖对培养大鼠颗粒细胞生成甾体激素的影响

目的研究中药紫草的水溶成分——紫草多糖对体外培养颗粒细胞的直接作用。方法分离出大鼠双侧卵巢的颗粒细胞进行体外培养。选取健康未成年雌性SD大鼠160只,其中60只按随机数字表法分为基础组和hCG组,分别采用不同浓度（0.00、0.30、0.75、1.50、3.00、7.50 g·L^-1）紫草多糖培养,每组5只;另100只按随机数字表法分为空白对照组、紫草多糖组、hCG组和hCG-紫草多糖组,分别进行不同时间（0、1、2、3、4 h）的培养,每组5只,共计100只。采用放射免疫测定法（RIA）测定不同实验组颗粒细胞培养液中孕酮（Progesterone,P4）和雌二醇（Estradiol,E2）的浓度。结果紫草多糖呈剂量依赖性地抑制hCG刺激下颗粒细胞P4和E2的生成,大剂量（大于3.00 g·L^-1）也抑制基础P4和E2的生成,紫草多糖主要使培养液中的P4含量降低,但大剂量也可使细胞内的P4含量降低。紫草多糖作用于颗粒细胞1 h后明显抑制hCG促P4生成的作用,但细胞洗涤后再培养,对hCG的反应性与对照组比较无明显变化。结论紫草多糖可直接作用于体外培养的颗粒细胞,抑制hCG促甾体激素生成的作用,该抑制作用起效时间短,并且是可逆的。     

Cellular proliferation rate and insulin-like growth factor binding protein (IGFBP)-2 and IGFBP-3 and estradiol receptor alpha expression in the mammary gland of dairy heifers naturally infected with gastrointestinal nematodes during development

Mammary ductal morphogenesis during prepuberty occurs mainly in response to insulin-like growth factor-1 (IGF-1) and estradiol stimulation. Dairy heifers infected with gastrointestinal nematodes have reduced IGF-1 levels, accompanied by reduced growth rate, delayed puberty onset, and lower parenchyma-stroma relationship in their mammary glands. Immunohistochemical studies were undertaken to determine variations in cell division rate, IGF-1 system components, and estradiol receptors (ESR) during peripubertal development in the mammary glands of antiparasitic-treated and untreated Holstein heifers naturally infected with gastrointestinal nematodes. Mammary biopsies were taken at 20, 30, 40, and 70 wk of age. Proliferating cell nuclear antigen immunolabeling, evident in nuclei, tended to be higher in the parenchyma of the glands from treated heifers than in those from untreated. Insulin-like growth factor binding proteins (IGFBP) type 2 and type 3 immunolabeling was cytoplasmic and was evident in stroma and parenchyma. The IGFBP2-labeled area was lower in treated than in untreated heifers. In the treated group, a maximal expression of this protein was seen at 40 wk of age, whereas in the untreated group the labeling remained constant. No differences were observed for IGFBP3 between treatment groups or during development. Immunolabeling for α ESR (ESR1) was evident in parenchymal nuclei and was higher in treated than in untreated heifers. In the treated group, ESR1 peaked at 30 wk of age and then decreased. These results demonstrate that the parasite burden in young heifers negatively influence mammary gland development, affecting cell division rate and parameters related to estradiol and IGF-1 signaling in the gland.     

Removal of bisphenol A and 17α-ethinyl estradiol from landfill leachate using single-walled carbon nanotubes

In this study, the adsorption of bisphenol A (BPA) and 17α-ethinyl estradiol (EE2) from landfill leachate onto single-walled carbon nanotubes (SWCNTs) was investigated. Different leachate solutions were prepared by altering the pH, ionic strength, and dissolved organic carbon (DOC) in the solutions to mimic the varying water conditions that occur in leachate during the various stages of waste decomposition. The youngest and oldest leachate solutions contained varying DOC and background chemistry and were represented by leachate Type A (pH = 5.0; DOC = 2500 mg/L; conductivity = 12,500 μS/cm; [Ca²⁺] = 1200 mg/L; [Mg²⁺] = 470 mg/L) and Type E (pH = 7.5; DOC = 250 mg/L; conductivity = 3250 μS/cm; [Ca²⁺] = 60 mg/L; [Mg²⁺] = 180 mg/L). These solutions were subsequently combined in different ratios to produce intermediate solutions, labeled B-D, to replicate time-dependent changes in leachate composition. Overall, a larger fraction of EE2 was removed as compared to BPA, consistent with its higher log K_OW value. The total removal of BPA and EE2 decreased in older leachate solutions, with the adsorptive capacity of SWCNTs decreasing in the order of leachate Type A > Type B > Type C > Type D > Type E. An increase in the pH from 3.5 to 11 decreased the adsorption of BPA by 22% in young leachate and by 10% in old leachate. The changes in pH did not affect the adsorption of EE2 in the young leachate, but did reduce adsorption by 32% in the old leachate. Adjusting the ionic strength using Na⁺ did not significantly impact adsorption, while increasing the concentration of Ca²⁺ resulted in a 12% increase in the adsorption of BPA and a 19% increase in the adsorption of EE2. DOC was revealed to be the most influential parameter in this study. In the presence of hydrophilic DOC, represented by glucose in this study, adsorption of the endocrine disrupting compounds (EDCs) onto the SWCNTs was not affected. In the absence of SWCNTs, hydrophobic DOC (i.e., humic acid) adsorbed 15-20% of BPA and EE2. However, when the humic acid and SWCNTs were both present, the overall adsorptive capacity of the SWCNTs was reduced. Hydrophobic (π-π electron donor-acceptor) interactions between the EDCs and the constituents in the leachate, as well as interactions between the SWCNTs and the EDCs, are proposed as potential adsorption mechanisms for BPA and EE2 onto SWCNTs.     

Cardiovascular,Metabolic and Inflammatory Changes after Ovariectomy and Estradiol Substitution in Hereditary Hypertriglyceridemic Rats

Background: If menopause is really independent risk factor for cardiovascular disease is still under debate. We studied if ovariectomy in the model of insulin resistance causes cardiovascular changes, to what extent are these changes reversible by estradiol substitution and if they are accompanied by changes in other organs and tissues. Methods: Hereditary hypertriglyceridemic female rats were divided into three groups: ovariectomized at 8th week (n = 6), ovariectomized with 17-β estradiol substitution (n = 6), and the sham group (n = 5). The strain of abdominal aorta measured by ultrasound, expression of vascular genes, weight and content of myocardium and also non-cardiac parameters were analyzed. Results: After ovariectomy, the strain of abdominal aorta, expression of nitric oxide synthase in abdominal aorta, relative weight of myocardium and of the left ventricle and circulating interleukin-6 decreased; these changes were reversed by estradiol substitution. Interestingly, the content of triglycerides in myocardium did not change after ovariectomy, but significantly increased after estradiol substitution while adiposity index did not change after ovariectomy, but significantly decreased after estradiol substitution. Conclusion: Vascular and cardiac parameters under study differed in their response to ovariectomy and estradiol substitution. This indicates different effects of ovariectomy and estradiol on different cardiovascular but also extracardiac structures.     

Yeast‐mediated stereo‐selective reduction of estrone by continuous cell culture with dual stirred tanks for product yield improvement

BACKGROUND: β‐Estradiol is an important hormone for the treatment of breast cancer and osteoporosis. β‐Estradiol can be produced via Saccharomyces cerevisiae mediated reduction of estrone. However, substrate inhibition and low production yield have been observed in the batch cell culture. RESULTS: An innovative continuous cell culture with dual stirred tanks in series was designed to solve the above problems. The growth medium was fed continuously to the incubation tank where the cells were incubated aerobically; the viable cells were then supplied continuously to the reaction tank in which the yeast‐mediated anaerobic reduction of estrone was performed with continuous feed of the substrate medium and continuous withdawal of the reaction cell culture. Thus, an increase in cell productivity from about 3‐fold to 7‐fold was obtained when compared with the batch cell culture. The β‐estradiol yield was improved to 64.8% on the second reaction day, accompanied by an accumulation of 12.9 mg β‐estradiol on the third reaction day. The yield was about 10% more and the accumulated recovery of β‐estradiol was 4.3‐fold better than with the batch cell culture. The diastereomeric excess value (%de) of β‐estradiol was more than 99%. CONCLUSION: A high product yield with excellent stereo‐selectivity was achieved in a short reaction period with the developed continuous cell culture and the dual stirred tank. Copyright © 2010 Society of Chemical Industry     

Reproductive performance of anestrous dairy cows treated with progesterone and estradiol benzoate

Anestrus is a major reproduction problem in pasture-based dairy operations that results in poorer reproductive outcomes than herdmates detected in estrus before the start of the seasonal breeding program. The objective of the current study was to assess a combined progesterone and estradiol benzoate treatment program including resynchrony with no treatment. Anestrous pasture-fed dairy cattle (n = 756) in 9 herds were blocked by herd and age and assigned within sequentially presented pairs of cows to be treated with an intravaginal progesterone-releasing insert for 8 d plus 2 mg of estradiol benzoate injected i.m. at insert insertion and 1 mg of estradiol benzoate injected 1 d after insert removal (d −1). Those cows detected in estrus from 0 to 3 d had a used progester-one-releasing insert reinserted for 6 d commencing on d 16 with 0.5 mg of estradiol benzoate injected i.m. 1 d after insert removal (treatment). The other cow within the pair was left as an untreated control (control). Treatment increased the risk of insemination and pregnancy by 28 d into the breeding program and resulted in conception 15 d earlier compared with controls. In contrast, treatment did not increase the risk of pregnancy after 56 d into the breeding program or at the end of the breeding season. It is concluded that treatment of anestrous dairy cattle with progesterone and estradiol benzoate combined with reinsertion of the progesterone insert resulted in earlier conception, but no difference in the final pregnancy rate compared with no treatment.     

Effect of time of artificial insemination and supplemental estradiol on reproduction of lactating dairy cows

Our objectives were to compare reproductive responses of dairy cows receiving timed artificial insemination (AI) either at 48 or 72 h after induction of luteolysis and supplemented or not with estradiol cypionate (ECP). Holstein cows (971) had their estrous cycles presynchronized with injections of PGF_2α at 37 and 51 d in milk (DIM) and then received an injection of GnRH at 64 DIM and an injection of PGF_2α at 71 DIM. Cows were then assigned to a 2 × 2 factorial randomized block experiment; cows in the CoSynch 48 h (CoS48) received a final injection of GnRH concurrent with timed AI 48 h after PGF_2α, whereas cows in the CoSynch 72 h (CoS72) received GnRH and timed AI 72 h after PGF_2α. Half of the cows in each CoSynch protocol received an injection of 1 mg of ECP 24 h after PGF_2α. Therefore, the 4 treatments were as follows: CoS48-NECP (n = 240), CoS72-NECP (n = 246), CoS48-ECP (n = 245), and CoS72-ECP (n = 240). Blood was sampled at 7 d before and at the first GnRH of the CoSynch from all cows for analysis of progesterone concentration in plasma. Cows were classified as anovular when progesterone was less than 1.0 ng/mL in both samples. Blood was also sampled during proestrus from a subset of 123 cows to measure concentrations of estradiol and at 7 d after timed AI to measure concentrations of progesterone. Ovaries from the same subset of 123 cows were examined by ultrasonography to determine ovulatory follicle diameter and incidence of ovulation. Pregnancy was diagnosed at 40 and 68 d after AI. Prevalence of cyclic cows was 72.4% and was similar among treatments. Concentrations of estradiol increased after ECP treatment and at 72 h of proestrus with CoS72. Pregnancy at 40 and 68 d after AI and pregnancy loss were not affected by timing of AI or supplemental ECP. Delaying timed AI to 72 h and supplementation with ECP increased the proportion of cows displaying estrus at AI, and cows detected in estrus had increased pregnancy per AI associated with improved ovulation and increased postovulatory progesterone concentration. These results indicate that extending the proestrus by delaying timed AI from 48 to 72 h plus supplemental ECP, despite increased expression of estrus at timed AI, did not improve reproductive performance of lactating dairy cows at first AI.     

基于表面等离子共振的全自动浓度检测系统

研制了以SpreetaTM传感器为核心,结合液体处理系统、信号检测与控制器、计算机分析处理软件等模块的小型表面等离子共振（SPR）检测系统.运用该系统,通过在传感器表面偶联牛血清蛋白（BSA）,采用直接检测法测量了不同浓度的BSA抗体溶液,得到了BSA抗体溶液的SPR传感图,建立了BSA抗体的浓度标准曲线.在传感器表面固定雌二醇（E2）,采用间接检测法测量了不同浓度的E2溶液,根据E2溶液SPR响应信号的变化量,建立了E2的浓度标准曲线.实验中系统的分辨率为10-5折射率单位（RIU）.对于实际的待测样品,只要测得其SPR信号的变化量,结合标准曲线就能够得出样品的浓度.