Cross-correlation image analysis for real-time single particle tracking.

Accurately measuring the translations of objects between images is essential in many fields, including biology, medicine, chemistry, and physics. One important application is tracking one or more particles by measuring their apparent displacements in a series of images. Popular methods, such as the center of mass, often require idealized scenarios to reach the shot noise limit of particle tracking and, therefore, are not generally applicable to multiple image types. More general methods, such as maximum likelihood estimation, reliably approach the shot noise limit, but are too computationally intense for use in real-time applications. These limitations are significant, as real-time, shot-noise-limited particle tracking is of paramount importance for feedback control systems. To fill this gap, we introduce a new cross-correlation-based algorithm that approaches shot-noise-limited displacement detection and a graphics processing unit-based implementation for real-time image analysis of a single particle.

Estrogen receptor alpha (ER-alpha) and beta (ER-beta) mRNAs in normal ovary, ovarian serous cystadenocarcinoma and ovarian cancer cell lines: down-regulation of ER-beta in neoplastic tissues.

The prognosis in ovarian carcinoma, the most lethal of the gynecologic neoplasms, is poor and has changed little in the last three decades. Only a small number respond to antiestrogen therapy, although the classic estrogen receptor, ER-alpha, has been identified in ovarian surface epithelium, from which approximately 90% of ovarian cancers originate. We have previously shown that ER-beta mRNA is most abundant in human fetal ovaries, suggesting that it might play an important role in ovarian development. Therefore, we investigated the mRNA levels of both ERs in normal ovaries, ovarian serous cystadenocarcinomas, granulosa cells from patients undergoing in vitro fertilization (IVF), the ovarian surface epithelium cell line IOSE-Van, and the ovarian cancer cell lines SKOV3, HEY and OCC1. Northern blots of normal and neoplastic ovaries were hybridized with an ER-beta riboprobe that spans the A/B domain. We detected two major hybridizing bands at approximately 8 and 10 kb. An RNase protection assay using the same probe revealed a single band of the expected size. Hybridizing the same blot with an ER-alpha riboprobe showed a strong hybridizing band at approximately 6.5 kb. In ovarian cancer samples, ER-beta mRNA level was decreased when compared to normal ovaries. Using 25 cycles of RT-PCR followed by Southern blotting, we found equal amounts of ER-alpha and -beta mRNAs in normal ovaries in all age groups from 33 to 75 years; however, in ovarian cancer tissue, the level of ER-alpha mRNA was similar or slightly higher, comparable to 10(3) to 10(4) copies of plasmid DNA, but ER-beta mRNA levels were markedly decreased. Granulosa cells from IVF patients expressed high levels of ER-beta mRNA. The OSE cell line expressed a low level of ER-alpha, detectable after 40 cycles of RT-PCR and no ER-beta mRNA. SKOV3 showed a low level of ER-alpha and -beta mRNAs, whereas OCC1 showed a low level of ER-beta and a relatively high level of ER-alpha. HEY did not contain detectable amounts of either ER after 40 cycles of RT-PCR. We found no evidence of differential splicing or major deletions in almost the entire coding region of ER-beta in either normal ovaries or tumor samples.

Tissue distribution of estrogen receptors alpha (ER-alpha) and beta (ER-beta) mRNA in the midgestational human fetus.

We compared the expression profiles of the mRNAs of both estrogen receptors, ER-alpha and the recently cloned ER-beta, in the midgestational human fetus by semiquantitative RT-PCR. ER-alpha was most abundant in the uterus, and smaller quantities were detected in the ovary, testis, skin and gut. High amounts of ER-beta mRNA were present in fetal ovaries, testes, adrenals and spleen. In these tissues, the levels of ER-beta mRNA were higher than ER-alpha. In the uterus, however, ER-alpha mRNA was more abundant, and ER-beta mRNA was expressed only moderately. ER-beta mRNA was present at moderate to low levels in the thymus, pituitary gland, skin, lung, kidney and brain cortex. In the course of our work, using the ER-beta primers on genomic DNA, an intron of 2468 bp in length, located between nt 222 and 223 in the A/B domain of ER-beta cDNA, was detected, cloned and sequenced. The study shows that the expression profile of the two ERs is different, and ER-beta is expressed in a variety of tissues during human fetal development, suggesting different, organ-specific roles for the two receptors.

Kallmann syndrome and associated malformation of the uterus.

Kallmann syndrome is a rare combination of hypogonadotropic hypogonadism due to hypothalamic insufficiency and anosmia. In both patients treated at our institution for infertility, a malformation of the uterus was noted: one patient had a unicornuate uterus, the other a uterus with a fundal hypoplasia and tubes of approximately 9 cm. It is not clear if the malformation is in association with Kallmann syndrome or purely coincidental. Given the known association of other malformations with Kallmann syndrome, we suppose that probably an underlying genetic defect having to do with organogenesis is the cause of the uterine malformations.

Postpartum periarticular hip abscess with later coxitis caused by group B streptococcus.

Intrapartum haematogenous spread of vaginal group B Streptococcus is rare, but it can lead to severe complications like abscesses, endocarditis and meningitis postpartum. We report a postpartum periarticular hip abscess caused by group B Streptococcus. Clinically it caused pain in the hip and a compression of the femoral nerve with motor and sensory component. Diagnosis was made by aspiration under computed tomography control. The only sign of infection was an increased sedimentation rate. After antibiotic treatment the symptoms disappeared and the abscess vanished, but it was reactivated 4 months after delivery, leading to arthritis of the hip joint. Another antibiotic treatment was administered for 8 weeks. 17 months postpartum the patient is well, but a development of a secondary coxarthrosis can not be excluded.

[Vulvar cancer. Bern university gynecologic clinic patients treated 1978 to 1987]. / Das Vulvakarzinom. An der Universitäts-Frauenklinik Bern von 1978 bis 1987 behandelte Fälle.

Between 1978 and 1987, 44 women with primary invasive carcinoma of the vulva were treated. The mean age of the patients was 69 years (24-91). The predominant histologic type was squamous cell carcinoma. Thirteen radical vulvectomies were performed. Various other vulvar operative procedures were performed on 27 cases and were followed up with or without radiation therapy. The 5-year survival rate was 80% in stage T1 patients, 58% in T2 and 40% in T3. Patients with negative lymph nodes had a survival rate of 79% after 5 years and those with positive nodes a survival rate of 40%. A modified radical vulvectomy was performed: to the Pary-Jones incision we use an additional groin incision. The advantage is better wound healing as compared to Way's technique.

Endometrial and placental protein markers and ovarian steroids in serum during in-vitro fertilization cycles.

The objective of this study was to find the earliest time at which it was possible to detect clinical pregnancy in an in-vitro fertilization (IVF) treatment cycle supported with human chorionic gonadotrophin (HCG), and also retrospectively to diagnose abnormal ovarian- or endometrium-related situations in failure cycles. Serum samples were taken in 41 IVF cycles at frequent intervals from the beginning of ovarian stimulation until menstrual bleeding occurred or a pregnancy was established. Concentrations of oestradiol, progesterone, placental protein 14 (PP14), pregnancy-specific beta 1-glycoprotein (SP1), and pregnancy-associated plasma protein A (PAPP-A) were determined in the serum samples using commercially available (steroid) or purpose-developed (protein) immunoassays. The cycles were retrospectively distributed into four outcome groups: (i) fertilization failure (FF, n = 8); (ii) implantation failure (IF, n = 10); (iii) 'interaction' (embryo-endometrium) cycle (IC, n = 14), and (iv) clinical pregnancy (CP, n = 9). The embryo-endometrium interaction was detected by a rise in SP1 in 23 cycles (70% of embryo transfers) at a time when endogenous HCG was still masked by external support. Early ('false') positive SP1 concentrations were observed in two out of eight and five out of 14 cases in groups FF and IC respectively, but never amongst the ongoing pregnancies (CP). PAPP-A did not distinguish pregnancy from the other outcomes. The PP14/progesterone ratio was lower, later in the cycle, in CP than in the other groups.(ABSTRACT TRUNCATED AT 250 WORDS)

[Homologous intrauterine insemination as a therapeutic possibility in andrologically-induced sterility]. / Die homologe intrauterine Insemination als Therapiemöglichkeit der andrologisch bedingten Sterilität.

In a two-years period, we treated 58 infertile couples for a total of 213 cycles with artificial homologous insemination (AIH). We considered a indication for AIH low sperm quality or a pathological postcoital test (PCT). Couples with idiopathic infertility did not receive AIH. If possible, AIH was performed by intrauterine insemination (IUI), following concentration of the motile sperms by "swim-up" method. Out of 213 cycles "swim-up", preparation could not be done in 46 cases (21.6%) because the initial sperm quality was below technical limits (sperm count below 1 Mio/ml). Out of 58 treated couples 4 became pregnant during AIH therapy (7%) among, them 2 couples conceiving twice after spontaneous early abortion. The total pregnancy rate per cycle was 2.8%. In the cycles after "swim-up", the monthly pregnancy rate through IUI was 3.6%. In the presence of andrological factors was the pregnancy rate only 0.9% compared to 6.8% in cases of pathological PCT with normal sperm quality. Thus, it can be concluded, that IUI is not successful on the basis of low sperm quality. Only in cases of unfavourable cervical factor expressed by a pathological PCT accompanied with a normal spermiogramm, moderate success can be expected.

Lasers in endoscopic surgery.

Lasers provide the endoscopic surgeon with a sophisticated tool for high precision cutting, destruction of tissue and coagulation. The laser allows efficient and time-saving operating techniques. Various laser systems emitting at different wavelengths are available. The absorption of the laser is determined by the tissue properties, such as water content or pigmentation, and varies strongly with the wavelength. Each laser offers advantages for the treatment of specific disorders. Further data from clinical studies are required to evaluate whether the advantages found in experimental studies are of significance in clinical use.

[Diagnosis of malignant uterine tumors by transvaginal ultrasound]. / Diagnostik maligner Uterustumoren durch die transvaginale Sonographie.

Two hundred and three non-pregnant women who underwent diagnostic dilatation and curettage or hysterectomy were included in a prospective study designed to assess the accuracy of transvaginal sonography in conforming or excluding the presence of uterine cancer. All preoperative sonographic findings were compared with the final histological diagnosis. The predictive value of sonographic evidence of malignancy was 71.1% (32/45), the predictive value of benign disease was 95.6% (151/158). Sensitivity was 82.1% (32/39) and specificity 92.1% (151/164). Although sensitivity and specificity increased as compared to an earlier study using transabdominal ultrasound, we recommend dilatation and curettage in women with postmenopausal bleeding.

Serum pregnancy-specific beta1-glycoprotein before embryo transfer is related to endometrial thickness and to outcome prognosis in women undergoing in-vitro fertilization treatment.

We have previously observed the repeated presence of low but detectable amounts of the trophoblast marker pregnancy-specific beta1-glycoprotein (SP1) in the serum of some women undergoing in-vitro fertilization (IVF) treatment around the time of oocyte retrieval. The occurrence of these signals seemed to be restricted to a defined group of patients which also showed a lower pregnancy success rate in a preliminary study. To test our hypothesis we have analysed 173 consecutive cycles leading to an embryo transfer. Fifty-four cycles (31%) had a serum SP1 level of at least 0.1 ng/ml between days embryo transfer -5 and embryo transfer (group A). Five pregnancies were obtained in this group (pregnancy rate = 9.3%), while in group B, defined by the absence of detectable SP1 before embryo transfer (119 cycles), 36 ongoing pregnancies were achieved (30.3%). Ten of the 41 pregnancies were achieved in 33 first-time non-pregnant patients undergoing further attempts during the study period. Again the pregnancy rate was higher in the first-time group B women (9/23 versus 1/10 for group A). Patients tended to remain in their groups A or B, the latter being associated with a better immediate as well as subsequent chance for pregnancy. Group A cycles had a significantly lower endometrial thickness two days before oocyte retrieval than group B (P = 0.0011). We postulate that the presence of an unknown, maternal and progesterone- or follicle stimulating hormone-independent factor in some patients could stimulate tonic ectopic SP1 synthesis and at the same time negatively influence endometrial development.

Detection of human papillomavirus in vulvar carcinoma. A study by in situ hybridisation.

Fourty-four specimens of invasive cancers of the vulva, including 38 primary invasive squamous carcinomas, were analysed by in situ hybridisation with biotinylated HPV 6/11, 16 and 18 DNA probes. Four (9%) of the 44 carcinomas were positive for HPV DNA: three (7%) for HPV 16 DNA and one (2%) for HPV 6/11 DNA. HPV DNA was found only in squamous carcinomas. Of the 38 primary squamous carcinomas, 11% were positive (8% HPV 16, 3% HPV 6/11). The overall 5-year survival was 48.7%, 48.5% for the squamous carcinomas and 50.0% for the HPV DNA positive patients.

Production of endometrial placental protein 14 and prolactin by cultured endometrial explants after collagenase and freeze/thaw treatment, and in response to progesterone.

OBJECTIVE: Investigation of methods for maintaining functional endometrial explants in culture after cryopreservation with or without previous enzymatic dispersion of stromal cells and epithelial glands. Such a standardized culture system is a requirement for the development of a non-invasive bioassay for embryo quality in in vitro fertilization programs, a method that will eventually measure endometrial response to embryo conditioned media. METHOD: Culture of mid-luteal phase endometrial biopsies, in the presence of [35S]methionine, with or without prior collagenase treatment and/or storage in liquid nitrogen in the presence of dimethyl sulfoxide. Determination of released de novo synthesized total protein by trichloroacetic acid precipitation of culture media. Measurement, after culture in absence and presence of progesterone, of prolactin and placental protein 14 (PP14) production by sensitive non-isotopic immunoassays. RESULTS: Production of prolactin, but not PP14, was increased by 200 nmol/l progesterone, 2-8-fold after 4 days and 1.5-700-fold after 7 days in culture. After limited collagenase treatment (but without separation of stromal cells from glands), both marker protein productions were similar compared to untreated explants; however, there was no significant stimulation of prolactin by progesterone. After freezing and thawing, production was markedly reduced, particularly from explants frozen following collagenase treatment. CONCLUSIONS: Both stromal and glandular viability are maintained after collagenase treatment but the response to progesterone is lost. Cryopreservation reduced prolactin and PP14 production in subsequent culture. Therefore, novel freezing protocols should be developed which preserve both endometrial structure and function.

CA-125 concentrations in the serum and pregnancy outcome in IVF cycles.

PURPOSE: CA-125 has been proposed as a potential marker for endometrial receptivity in assisted reproduction. This study was designed to evaluate whether the levels of CA-125 in the serum of patients undergoing IVF-embryo transfer (ET) is correlated with the outcome. METHODS: Levels of serum CA-125 were measured on the day before and on the day of human chorionic gonadotropin (hCG) administration, ovum pickup (OPU), and ET in 74 patients undergoing 100 IVF cycles between January 1994 and March 1995. Patients were treated with a midluteal-phase gonadotropin-releasing hormone (GnRH) agonist protocol and follicular-phase human menopausal gonadotropin. RESULTS: One hundred oocyte retrievals resulted in 91 ETs, and 22 clinical pregnancies (22%/OPU and 24.2%/ET). The live-born rate was 21%/OPU and 23.1%/ET. Neither the CA-125 serum levels nor their increase from the day of hCG until the day of ET showed any prognostic significance to the outcome of IVF, and they were not correlated with the endometrium thickness or the number of oocytes retrieved or fertilized. CONCLUSIONS: The CA-125 serum levels in conventional IVF cycles were not correlated with the IVF outcome and yielded no prognostic information in a GnRH agonist down-regulation protocol.

Oestrogen receptor (ER)-alpha and ER-beta isoforms in normal endometrial and endometriosis-derived stromal cells.

Several investigators have noted that hormone-dependent development of endometriosis implants lags behind that of simultaneously analysed eutopic endometrium. With the recent discovery of the oestrogen receptor-beta (ER-beta) isoform, the aim of this study was to investigate whether differences in the expression of ER-alpha and ER-beta might explain this observation. mRNA transcripts from endometrial stromal cells isolated from normal endometrium (NE) and from endometriomas (EI) were analysed using a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) technique. RT-PCR and Southern blot analyses of the two ER isoforms indicated that NE and EI stromal cells predominantly express ER-alpha mRNA, however the relative concentrations of ER isoform mRNA transcripts differed between the two cell types. Steady-state ER-alpha:ER-beta mRNA ratios were 15.5 +/- 2.8 and 5.2 +/- 0.9 respectively for NE and EI cells (P = 0.02). NE and EI stromal cells expressed ER proteins with similar Kd ( approximately 0.9 nM) and densities ( approximately 24 500 binding sites/cell) respectively. Functional ER expression was indicated by an increase in progesterone receptor concentrations of approximately 60% (P = 0.03) after incubation with 10 nM oestradiol. We postulate that differential transcript processing, ligand specificity and biological actions of the ER-alpha and -beta isoforms may influence differential growth responses in normal and ectopic endometrium.