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- Appelblad, Patrik, et al.
(författare)
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Determination of C-21 ketosteroids in serum using trifluoromethanesulfonic acid catalyzed precolumn dansylation and 1,1’-oxalyldiimidazole postcolumn peroxyoxalate chemiluminescence detection
- 1998
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Ingår i: Analytical Chemistry. - Washington : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 70:23, s. 5002-5009
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Tidskriftsartikel (refereegranskat)abstract
- A new procedure for the quantitation of C-21 ketosteroids using trifluoromethanesulfonic acid-catalyzed precolumn dansylation and coupled column liquid chromatographic separation, followed by postcolumn 1,1‘-oxalyldiimidazole peroxyoxalate chemiluminescence detection is presented. In the simultaneous optimization of chromatographic resolution and chemiluminescence intensity, a coupled column chromatographic system and a stopped-flow system were used. An eluent containing 20 mM phosphate buffer at pH 6.7 accomplished an efficient separation of 3α-hydroxy-5β-pregnan-20-one from a mixture containing 10 C-21 ketosteroids. Phosphate buffer also proved to be the most advantageous, among the six buffers tested, for sensitive detection. Experimental design and multivariate data analysis were used to characterize and optimize the postcolumn reaction chemistry in the chromatographic system. A valid full factorial design with excellent predictability showed that the flow rates for both 1,1‘-oxalyldiimidazole and hydrogen peroxide were the factors most strongly affecting the sensitivity of the system. The theoretical plate numbers were above 11 000 for all 10 dansylated ketosteroids. The 3σ detection limit estimated from 3α-hydroxy-5β-pregnan-20-one calibration curve data was 1.6 pmol (n = 4, 125 μL injected) and spiked serum containing 0−74 pmol of this compound showed overall recoveries of 73 ± 9% (n = 12). Quantitation of 3α-hydroxy-5β-pregnan-20-one was finally carried out on 45 serum samples and the results compared to those from a radioimmunoassay (RIA) method. The data acquired with the procedure described in this work compare well with the results from RIA, which confirms the reliability of the new analytical procedure.
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| 2. |
- Björn, Inger, 1953-, et al.
(författare)
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Drug related negative side effects is a common reason for poor compliance in hormone replacement therapy
- 1999
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Ingår i: Maturitas. - : Elsevier. - 0378-5122 .- 1873-4111. ; 32:2, s. 77-86
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Tidskriftsartikel (refereegranskat)abstract
- Objectives: The reasons for poor compliance with hormone replacement therapy (HRT) and, in particular drug-related reasons, have not yet been fully elucidated. In this study, a cohort of peri- or postmenopausal women—mainly workers from a small town and surrounding rural area—was studied. The aim of the study was to investigate why some women never start or discontinue HRT, even when great effort has been made to inform and fulfill the demands of the patient. Methods: All women who were given a HRT prescription at a gynecological practice between September 1991 and December 1992 participated in a longitudinal study. A written questionnaire was mailed to these patients in 1996. Data from the questionnaire was supplemented with information from the medical records. Care of patients included initial information, follow-up within 4 months, yearly visits supplemented with contacts on demand. Results: 356 women received the questionnaire, among which 92% replied. A total of 2% never started HRT. Seventy-five percent continued the therapy for more than 3 years. Reasons for discontinuing HRT were negative side-effects (35%), desire to find out if climacteric symptoms had ended (26%), fear of cancer and thrombosis (25%), weariness of bleeding (19%) and a wish to deal with the problems ‘naturally’ (15%). Conclusions: Compliance with HRT can be high if adequate information is given and follow-ups are made. The main reason for poor compliance was negative side-effects, most likely progestin-related. The results of this study suggest that the future challenge will be to minimize negative side-effects of HRT.
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| 3. |
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| 4. |
- Ingvarsson, Magdalena, et al.
(författare)
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Isolation and culture of ovarian tumour cells, cytological and cell survival evaluation
- 1999
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Ingår i: Anticancer Research. - 0250-7005 .- 1791-7530. ; 19:6B, s. 5069-5073
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Tidskriftsartikel (refereegranskat)abstract
- The purpose of this study was to evaluate the reproducibility and reliability of the fluorometric microculture cytotoxicity assay (FMCA). Emphasis was placed on obtaining pure tumour cell cultures which were subjected to careful cytological evaluation. Preparations of 39 ovarian tumours, malignant, borderline and benign were made, of which 37 were successfully cultured. In 34 of the 37 tumour cell cultures, the epithelial cell fraction was > 90%, and in 30 of 39 cultures the epithelial cell fraction was > 95%. Transportation within 24 h and the 72 h incubation did not change the yield or epithelial cell fraction. There was a linear relationship between fluorescence and the number of viable cells. The fluorescence increased with time, making only comparisons within each assay plate possible. The sensitivity of the method makes it possible to perform many analyses on a small amount of material. The method also makes it possible to study cells derived from all stages of the disease, including benign tumours.
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| 5. |
- Löfgren, Mats, et al.
(författare)
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High progesterone is related to effective human labor : study of serum progesterone and 5α-pregnane-3,20-dione in normal and abnormal deliveries
- 1997
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Ingår i: Acta Obstetricia et Gynecologica Scandinavica. - 0001-6349 .- 1600-0412. ; 76:5, s. 423-430
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Tidskriftsartikel (refereegranskat)abstract
- Background. The role of progesterone levels during human labor is unclear. Objective. To investigate serum concentrations of progesterone and 5α-pregnane-3,20-dione in normal and abnormal deliveries. Methods. Venous and umbilical cord serum samples were collected from 108 parturient women. In a further 49 deliveries, arterial and venous umbilical cord sera were collected separately. The concentrations of progesterone and 5opregnane-3,20-dione were determined by radioimmunoassay. The delivery modes studied were: elective cesarean section; oxytocin-resistant dystocia; normal but induced delivery, and normal spontaneous delivery. Results. Progesterone concentrations in maternal and umbilical serum were higher following normal labor than after dystocia (p<0.005) and elective cesarean section (p<0.005). The maternal and umbilical progesterone concentrations in dystocia and elective cesarean section were between 77–43 % of those in normal labor. The concentrations did not vary between gestational weeks 37 and 42, within the different modes of delivery. The 5α-pregnane-3,20-dione serum concentration in the fetal compartment was twice that in the maternal compartment (p<0.001); its concentration in venous umbilical serum was higher than in corresponding arterial samples (p<0.001). No distinct differences in the 5α-pregnane-3,20-dione serum concentration were found with regard to parity or mode of delivery. Conclusion. High progesterone concentrations during parturition appear to be related to effective labor. The findings support results from in vitro experiments on human term myometrium. Read More: http://informahealthcare.com/doi/abs/10.3109/00016349709047823
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| 6. |
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| 7. |
- Ottander, Ulrika, et al.
(författare)
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Compartmentalization of human chorionic gonadotrophin sensitivity and luteinizing hormone receptor mRNA in different subtypes of the human corpus luteum
- 1997
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Ingår i: Human Reproduction. - : Oxford University Press (OUP). - 0268-1161 .- 1460-2350. ; 12:5, s. 1037-1042
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Tidskriftsartikel (refereegranskat)abstract
- The relationship was investigated between different ultrasonographically defined subtypes of the human corpus luteum and progesterone production. Twenty-one women in the mid-luteal phase who underwent laparotomy for benign uterine conditions volunteered for this study. The corpus luteum was identified by preoperative ultrasound and classified into four types according to earlier established criteria, where types a and c were centrally hypoechoic, types b and d were centrally echogenic, types a and b had thin surrounding 'walls' (<3 mm) and types c and d had thick walls (<3 mm). After luteectomy, the theca externa capsule was removed and tissue from directly beneath the surface ('peripheral region') and the layer immediately beneath ('inner region') minced into 4-6 mg pieces. Following preincubation, pieces were incubated for 3 h at 37 degrees C in HEPES-minimal essential medium buffer with or without human chorionic gonadotrophin (HCG; 10 IU/ml), and subsequently progesterone accumulation in the medium was determined by radioimmunoassay. The highest progesterone production was consistently seen in the peripheral region. Type a had a significantly (P > 0.01) lower progesterone production (3.2 +/- 1.5 nmol/g tissue wet weight, mean +/- SEM, n = 4) than that of types b, c and d (17.7 +/- 3.5 nmol/g tissue wet weight, n = 9). All types responded to HCG with an almost two-fold increase in progesterone production. However, the maximal progesterone produced following stimulation by HCG in the type a corpus luteum was <50% of the basal (unstimulated) progesterone synthesis of any other type of corpus luteum. Using in-situ hybridization, with a primate RNA probe complementary to the region coding the extracellular part of the luteinizing hormone (LH) receptor, a highly localized expression of LH receptor mRNA to the peripheral region was found. Negligible or low levels of expression were found in the theca externa capsule and the inner region. No obvious correlations between the different subtypes of corpora lutea and LH receptor mRNA expression were seen. Thus, the ultrasonographic detection of a thin-walled and centrally hypoechoic corpus luteum correlates well with reduced progesterone secretion. The underlying cellular mechanism does not appear to involve a diminished sensitivity to the gonadotrophic stimulation by LH or HCG.
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| 8. |
- Smans, Karine A, et al.
(författare)
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Bispecific antibody-mediated lysis of primary cultures of ovarian carcinoma cells using multiple target antigens
- 1999
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Ingår i: International Journal of Cancer. - 0020-7136 .- 1097-0215. ; 83:2, s. 270-277
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Tidskriftsartikel (refereegranskat)abstract
- We have shown previously that a bispecific antibody (BsAb) directed against both germ-cell alkaline phosphatase (GCAP) and the CD3 complex on mouse T cells could effectively eliminate GCAP-positive tumor cells in vivo using an immunocompetent mouse model. However, some GCAP-negative tumor cells were still able to grow, suggesting that BsAb therapy, when used in a clinical setting, could benefit from targeting several tumor markers to prevent outgrowth of tumor cells lacking a targeted marker. To test this hypothesis, we developed an in vitro model based on primary human ovarian carcinoma (OC) cultures and BsAbs directed against human T cells and several tumor markers [placental alkaline phosphatase (PLAP), GCAP, folate-binding protein (FBP) and CA19.9]. OC cells, isolated from primary tumors, were co-cultured with human peripheral blood mononuclear cells in the presence or absence of various concentrations of BsAbs against PLAP/GCAP, FBP and CA19.9 administered separately or in combination. Results derived from 18 primary OC samples showed that the combination treatment was better than or equally effective as the best single BsAB treatment in 60% of cases. Sometimes targeting FBP, PLAP/GCAP or CA19.9 alone was superior to targeting all simultaneously. Combining each BsAb with a low dose of IL-2 was always beneficial. These results indicate that before using a specific BsAb in the clinic, it is important to determine the optimal BsAb for each patient using this in vitro assay on cells from the removed tumor mass.
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