[Show abstract][Hide abstract] ABSTRACT: To measure and compare placental mRNA expression in the maternal circulation among women with intrauterine and ectopic pregnancies.
Plasma was collected from patients in early pregnancy at risk of ectopic pregnancy. Clinical information was prospectively collected and entered into a dedicated database. mRNA was isolated from maternal plasma and quantitative RT-PCR was performed to measure mRNA for human gonadotropin (hCG) and human placental lactogen (hPL). GAPDH mRNA expression was used as an internal control.
Twelve women with ectopic pregnancy and 13 women with intrauterine pregnancy were enrolled. Patients with ectopic pregnancy were 6 times more likely to have undetectable levels of hPL mRNA (relative risk [RR] 6.36; 95% confidence interval [CI], 1.70-23.20; P<0.01). They were also 8 times more likely to have undetectable levels of hCG mRNA (RR 8.64, 95% CI, 1.30-57.10; P<0.01). mRNA copy numbers for hPL and hCG (normalized by GAPDH) were significantly lower in the ectopic group than in the intrauterine group.
Placental mRNA is present in the maternal circulation in significantly lower copies in cases of ectopic pregnancy compared with cases of intrauterine pregnancy. Measurement of placental mRNA in the maternal circulation may help to distinguish between intrauterine and ectopic pregnancies.
International journal of gynaecology and obstetrics: the official organ of the International Federation of Gynaecology and Obstetrics 02/2012; 117(2):131-3. · 1.41 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Array-based comparative genomic hybridization (a-CGH) is a promising tool for clinical genomic studies. However, pre-analytical sample preparation methods have not been fully evaluated for this purpose. Parallel sections of normal male human skin biopsy samples were collected and immediately immersed in saline, formalin and a molecular fixative for 8, 12 and 24 h. Genomic DNA was isolated from the samples and subjected to amplification and labeling. Labeled samples were then co-hybridized with normal reference female DNA to Agilent oligonucleotide-based a-CGH 44k slides. Pre-analytic parameters such as DNA yield, quality of genomic DNA and labeling efficacy were evaluated. Also microarray analytical variables, including the feature signal intensity, data distribution dynamic range, signal to noise ratio and background intensity levels were assessed for data quality. DNA yield and quality of genomic DNA--as evaluated by spectrophotometry and gel electrophoresis--were similar for fresh and molecular fixative-exposed samples. In addition, labeling efficacy of dye incorporation was not drastically different. There was no difference between fresh and molecular fixative material comparing scan parameters and stem plot analysis of a-CGH result. Formalin-fixed samples, on the other hand, showed various errors such as oversaturation, non-uniformity in replicates, and decreased signal to noise ratio. Overall, the a-CGH result of formalin samples was not interpretable. DNA extracted from formalin-fixed tissue samples is not suitable for oligonucleotide-based a-CGH studies. On the other hand, the molecular fixative preserves tissue DNA similar to its fresh state with no discernable analytical differences.
Archives for Dermatological Research 10/2007; 299(7):353-7. · 2.71 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study aimed to measure the effects of peroxisome proliferator-activated receptor-δ (PPARδ) agonist GW501516 (GW) and zinc sulfate (ZS) on ovariectomized rats' vaginal histomorphology and collagen expression.
Two weeks after ovariectomy, rats received daily treatment with vaginal suppositories containing placebo, ZS, GW, ZS with GW, or estradiol for 2 weeks. Macroscopic measurements were taken and the midsection of the vagina was used for histology. Immunofluorescence was performed with antibodies against collagen I, III, and anti-actin or collagen I and V and anti-actin. Gene expression analysis of 3 collagen genes was performed by qRT-PCR.
Macroscopic measurements revealed that the genital hiatus was narrower in the ZS and ZS with GW groups, and the vagina was significantly longer in the animals treated with GW, ZS with GW, and estradiol compared to the placebo group. Microscopic measurements of the vaginal layers showed that the lamina propria and the vaginal muscularis were significantly thicker in the ZS and ZS with GW group compared to the placebo.The ratio of vaginal Col1a1/Col3a1 mRNA expression was significantly up-regulated by ZS with GW compared to placebo, whereas the ratio of vaginal Col1a1/Col5a1 expression was significantly up-regulated by ZS, GW, and ZS with GW. The ratio of vaginal collagen I/III protein expression was significantly up-regulated by ZS and ZS with GW, whereas the ratio of vaginal collagen I/V expression was significantly up-regulated by estradiol, ZS, and ZS with GW compared to control.
Vaginal suppositories containing zinc and PPARδ agonist significantly altered the vagina of ovariectomized rats.
Journal of Pelvic Medicine and Surgery 19(3):126-31.