Antalet vindkraftverk, bade i Sverige och i varlden, okar fran ar till ar samtidigt som de blir allt hogre. Utvecklingen mot hogre torn och kraftigare turbiner kraver allt storre fundament for att ...
Secreted phosphoprotein 1 (SPP1, osteopontin) is an extracellular matrix (ECM) protein that binds integrins to affect cell-cell and cell-matrix interactions including adhesion, migration, proliferation and survival. Secreted-protein-acidic-and-rich-in-cysteine (SPARC, osteonectin) is a calcium-binding matricellular glycoprotein known to regulate ECM interaction during tissue remodeling and repair. We developed a Spp1-/-/Sparc-/- double null mouse in which 50% fewer pups are present at birth than are found in utero on days 18-20 of gestation due to postpartum maternal cannibalization of the pups, suggesting that these pups are stillborn or exhibit severe birth defects. Indeed, hemorrhages were observed prominently in lower right limbs and distal region of the tail in many embryos; in a few embryos, hemorrhages were present the intestine and right foreheads. In order to gain insight into the developmental abnormalities and decreased litter sizes seen in double null litters, we evaluated the expression of both Spp1 and Sparc in the mouse uterus throughout pregnancy. Ten-week old outbred CD-1 female mice were mated to an intact male of the same strain (copulatory plug = Day 1 of gestation). Pregnant females were sacrificed and uteri collected on Day 4, 4.5, 5, 9, 10, 11, 12, and 16 of pregnancy. Cell-specific expression of Spp1 and Sparc mRNA in serial sections of mouse uteri was determined by radioactive in situ hybridization analysis. On Day 4 of pregnancy, Spp1 was expressed in central cells of the inner cell mass (ICM) of the embryo, in scattered immune cells in uterine stroma, and in luminal epithelium (LE) directly contacting the embryo within the implantation chamber. LE expression within the implantation chamber was absent by Days 4.5-5 of pregnancy, but remained in LE adjacent to the implantation chamber and in interimplantation sites, suggesting that Spp1 may be involved in adhesion during the initial phase of embryo attachment to the LE, but subsequently must be down-regulated at sites of attachment to allow embryo invasion. Sparc was expressed diffusely in the uterine stroma, in peripheral cells of the ICM, and was not expressed in LE through Days 4.5-5. When we utilized a model of delayed implantation to determine the effect of nidatory estrogen, localization of Spp1 and Sparc was identical to that of pregnancy. On Day 9 of gestation, Spp1 was expressed by uterine natural killer (NK) cells and LE, whereas Sparc was expressed by decidual cells and LE. On Day 10, NK cells and LE continued to express Spp1, while Sparc was localized to the stroma directly beneath the LE. On Day 12, Spp1 was also expressed by trophoblast giant cells, and Sparc was expressed in the Reichert's Membrane adjacent to the trophoblast giant cells. By Day 16 of pregnancy, both Spp1 and Sparc were expressed in adjacent but non-overlapping regions of developing fetal bone and cartilage. Because Spp1 and Sparc often have antagonistic roles during tissue remodeling and are localized to adjacent but different cell types during mouse pregnancy, we hypothesize that simultaneous deletion of both Spp1 and Sparc leads to disruption of cell-cell, cell-matrix and inter-tissue communication essential to normal conceptus development that is not observed when the genes are deleted individually. This study was supported by NIH grants R01 CA90920 and RO1 CA137091. (platform)
Endometrium in polycystic ovary syndrome (PCOS) presents altered gene expression indicating progesterone resistance and predisposing to reduced endometrial receptivity and endometrial cancer.We hypothesized that an altered endocrine/metabolic environment in PCOS may result in an endometrial "disease phenotype" affecting the gene expression of different endometrial cell populations, including stem cells and their differentiated progeny.This was a prospective study conducted at an academic medical center.Proliferative-phase endometrium was obtained from 6 overweight/obese PCOS (National Institutes of Health criteria) and 6 overweight/obese controls. Microarray analysis was performed on fluorescence-activated cell sorting-isolated endometrial epithelial cells (eEPs), endothelial cells, stromal fibroblasts (eSFs), and mesenchymal stem cells (eMSCs). Gene expression data were validated using microfluidic quantitative RT-PCR and immunohistochemistry.The comparison between eEP(PCOS) and eEP(Ctrl) showed dysregulation of inflammatory genes and genes with oncogenic potential (CCL2, IL-6, ORM1, TNAIFP6, SFRP4, SPARC). eSF(PCOS) and eSF(Ctrl) showed up-regulation of inflammatory genes (C4A/B, CCL2, ICAM1, TNFAIP3). Similarly, in eMSC(PCOS) vs eMSC(Ctrl), the most up-regulated genes were related to inflammation and cancer (IL-8, ICAM1, SPRR3, LCN2). Immunohistochemistry scoring showed increased expression of CCL2 in eEP(PCOS) and eSF(PCOS) compared with eEP(Ctrl) and eSF(Ctrl) and IL-6 in eEP(PCOS) compared with eEP(Ctrl).Isolated endometrial cell populations in women with PCOS showed altered gene expression revealing inflammation and prooncogenic changes, independent of body mass index, especially in eEP(PCOS) and eMSC(PCOS), compared with controls. The study reveals an endometrial disease phenotype in women with PCOS with potential negative effects on endometrial function and long-term health.
We reported that supplementing 0.8% L-arginine (Arg) to the diet of gilts between Days 0 and 25 of gestation reduced the number of corpora lutea and embryonic survival. This study determined the effects of dietary Arg supplementation between Days 14 and 25 of gestation on survival and development of conceptuses (embryo/fetus and associated membranes) in gilts. Gilts were checked daily for estrus with boars and mated at onset of the second estrus and 12 h later. Three fertile boars of similar genetic background were used randomly for breeding to minimize boar effects. The day of onset of estrus and breeding was defined as Day 0 of gestation. Between Days 14 and 25 of gestation, gilts (n = 15 per treatment) were housed individually and fed twice daily 1 kg of a corn- and soybean meal-based diet (2 kg/day) supplemented with 0.0%, 0.4%, or 0.8% Arg. All diets were made isonitrogenous by addition of L-alanine. At Day 25 of gestation, gilts were hysterectomized to obtain conceptuses. There were no differences (P > 0.05) in total number of conceptuses, number of corpora lutea, total fetal weight, total volume of allantoic fluid, concentrations of progesterone and estrogen in maternal plasma, or total amounts of progesterone and estrogen in allantoic fluid among the three treatment groups of gilts. Dietary supplementation with 0.4% or 0.8% Arg increased (P < 0.01) the concentration of Arg in maternal plasma, total volume of amniotic fluid, and the vascularity of chorionic and allantoic membranes, compared with the control group. The average rates of embryonic mortality were 24.7%, 11.2%, and 10.1% in the 0.0%, 0.4% and 0.8% Arg groups, respectively (pooled SEM = 1.9). The average numbers of viable conceptuses per litter were 10.5, 12.7, and 12.2 in the 0.0%, 0.4% and 0.8% Arg groups, respectively (pooled SEM = 0.4). Reproductive performance did not differ (P > 0.05) between the 0.4% Arg and 0.8% Arg groups. However, in comparison with the control group, supplementation with 0.4% or 0.8% Arg reduced (P < 0.01) embryonic mortality by 55% to 59%, while increasing (P < 0.05) the number of viable conceptuses per litter by approximately two. These results indicate that supplementation with 0.4% or 0.8% Arg between Days 14 and 25 of gestation can enhance embryonic/fetal survival in gilts. (Supported by AFRI Grant No. 2008-35203-19120 from the USDA National Institute of Food and Agriculture and by the National Natural Science Foundation of China Grant No. 30810103902). (platform)
Conceptus implantation involves pregnancy-specific alterations in extracellular matrix at the conceptus-maternal interface. Secreted phosphoprotein 1 (SPP1, osteopontin) is induced just before implantation and is present at the conceptus-maternal interface in mammals. In the present study, we investigated mechanisms by which SPP1 facilitates porcine conceptus and uterine luminal epithelial cell attachment. Native bovine milk and wild-type rat recombinant SPP1 stimulated trophectoderm cell migration. Bovine milk SPP1, ovine uterine SPP1, and recombinant wild-type, but not mutated, rat SPP1 promoted dose- and cation-dependent attachment of porcine trophectoderm and uterine luminal epithelial cells, which was markedly reduced in the presence of a linear Arg-Gly-Asp integrin-blocking peptide. Affinity chromatography and immunoprecipitation experiments revealed direct binding of alpha v beta 6 trophectoderm and alpha v beta 3 uterine epithelial cell integrins to SPP1. Immunofluorescence microscopy using SPP1-coated microspheres revealed colocalization of the alpha v integrin subunit and talin at focal adhesions as well as at the apical domain of trophectoderm cells. Similarly, immunofluorescence staining of implantation sites in frozen gravid uterine cross sections localized SPP1 and alpha v integrin to the apical surfaces of trophectoderm and luminal epithelium and beta 3 integrin to the apical surface of luminal epithelium. To our knowledge, the present study is the first to demonstrate functionally that SPP1 directly binds specific integrins to promote trophectoderm cell migration and attachment to luminal epithelium that may be critical to conceptus elongation and implantation.
Between days 0 and 24 of gestation, 10 gilts per treatment were individually fed twice daily 1 kg of a corn‐ and soybean meal‐based diet supplemented with 0.0%, 0.4%, or 0.8% L‐arginine (Arg). The diets were made isonitrogenous by addition of appropriate amounts of L‐alanine (Ala). At day 25 of gestation, 30 min after consumption of Ala or Arg, gilts were hysterectomized to obtain conceptuses. Supplementation with 0.4% or 0.8% Arg increased (P<0.05) concentrations of Arg in maternal plasma (64% and 98%, respectively) as well as the vascularity of chorionic and allantoic membranes, compared with the control group. Numbers of corpora lutea (CL) and fetuses, placental and fetal weights, and embryonic mortality did not differ (P>0.05) between the 0.4% Arg and control groups. However, supplementation with 0.8% Arg decreased (P<0.05) uterine weight (−20%), total numbers of fetuses (−24%) and CL (−17%), total fetal weight (−34%), total volumes of allantoic and amniotic fluids (−34 to 42%), concentrations of progesterone in maternal plasma (−33%) as well as total amounts of progesterone (−35%), estrone (−40%), and estrone sulfate (−37%) in allantoic fluid, compared to the control group. Thus, supplementation with 0.8% Arg between days 0 and 25 of gestation, while enhancing placental vascularity, adversely affects the reproductive performance of gilts. Supported by USDA grants and TAMU Faculty of Nutrition. Grant Funding Source : USDA
Objective To determine whether increasing the dose of ulipristal acetate (UPA)-containing emergency contraception (EC) improves pharmacodynamic outcomes in individuals with obesity. Study design We enrolled healthy, regularly-cycling, confirmed ovulatory, reproductive-age individuals with body mass index (BMI) >30 kg/m 2 and weight >80 kg in a randomised crossover study. We monitored participants with transvaginal ultrasound and blood sampling for progesterone, luteinising hormone (LH), and estradiol every other day until a dominant follicle measuring >15 mm was visualised. At that point, participants received either oral UPA EC 30 mg or 60 mg and returned for daily monitoring up to 7 days. After a no treatment washout cycle, participants returned for a second monitored cycle and received the other UPA dose. Our primary outcome was the proportion of subjects with no follicle rupture 5 days post-dosing (yes/no). For reference, we also enrolled a control group with BMI <25 kg/m 2 and weight <80 kg who received UPA EC 30 mg during a single cycle. We also obtained blood samples for pharmacokinetic parameters for UPA and its active metabolite, N -monodemethyl-UPA (NDM-UPA) as an optional substudy. Results We enrolled a total of 52 participants with BMI >30 kg/m 2 and 12 controls, with the following cycles completed: 12 controls, 49 UPA 30 mg, and 46 UPA 60 mg. The entire cohort demographics were a mean (SD) age of 29.8 (3.4) years and BMI by group: controls 22.5 (1.4) kg/m 2 , group 1 37.9 (6.7) kg/m 2 , and group 2 39.3 (5.4) kg/m 2 . All 12 (100%) of controls had a delay of at least 5 days for follicle rupture. Among the high BMI group, dosing groups (UPA EC 30 mg vs 60 mg) were similar in the proportion of cycles without follicle rupture over 5 days post-UPA dosing (UPA 30 mg: 47/49 (96%), UPA 60 mg: 42/46 (91%), Fisher’s exact test p=0.43). However, after excluding cycles where dosing occurred too late (after LH surge), a delay of at least 5 days occurred in all participants at both doses. The 60 mg UPA dose resulted in a twofold increase in maximum observed concentration and the area under the curve of both UPA and NDM-UPA levels compared with 30 mg. Conclusion A standard 30 mg dose of UPA is sufficient to delay ovulation regardless of BMI or weight. Results of our study do not support dose adjustment for body size.
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