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Expression of the vascular endothelial growth factors B and C and their receptors in human endometrium during the menstrual cycle.

BACKGROUND: Endometrial angiogenesis is important for bleeding control and fertility, and is believed to be regulated by angiogenic growth factors under the influence of ovarian steroids. OBJECTIVE: To document the pattern of expression of the two angiogenic growth factors, vascular endothelial growth factor (VEGF)-B and VEGF-C, and of their receptors, VEGFR-1, VEGFR-2, and VEGFR-3, in order to investigate their possible role in the regulation of angiogenesis in normal cycling human endometrium. MATERIAL AND METHODS: Cryosections of endometrial biopsy specimens obtained from 17 healthy women before laparoscopic sterilization were examined by immunohistochemistry. RESULTS: Vascular endothelial growth factor and VEGF-C and their receptors were all expressed in and around endometrial blood vessels, with no obvious menstrual cycle-dependent changes in expression except for VEGFR-2, which showed stronger expression during the early secretary phase. Stromal blood vessel density did not change significantly during the menstrual cycle. CONCLUSION: The presence and pattern of expression of these angiogenic growth factors and their receptors suggest that they participate in the regulation of normal endometrial angiogenesis.

Endometrium↗

[Ultrasound diagnosis of focal intrauterine lesions].

OBJECTIVE: To compare the efficacy of the conventional grey-scale real time ultrasound, color Doppler ultrasound (color flow mapping) and hysteroscopy in the diagnosis of circumscript intrauterine lesions, such as endometrial polyps and submucosal myomas. METHODS: Preoperative transvaginal ultrasound data of 124 patients with focal intrauterine lesions, as confirmed by operative hysteroscopy, were elaborated. Using grey-scale ultrasound the detailed endometrial morphology, with color Doppler the vascularization pattern of the endometrium was examined. The ultrasound diagnosis, the hysteroscopic diagnosis and the histological diagnosis were matched and the sensitivity of conventional grey-scale ultrasound, color Doppler ultrasound and hysteroscopy were calculated. Age, menstrual and fertility status, medication and symptoms (abnormal bleeding) of patients were also evaluated. RESULTS: Of the 124 removed lesions the histological examination confirmed 114 endometrial polyps and 10 leiomyomas. Conventional grey-scale ultrasound was performed at 79 patients (TVS group), while 45 patient were examined by color Doppler ultrasound (TVCD group). In the TVS group the intrauterine lesion was detected in 59 cases, and the origin was also correctly diagnosed in 48 cases. In the TVCD group 42 lesions were detected, and the ultrasound diagnosis was consistent with the histological diagnosis in 41 cases. At hysteroscopy 118 endometrial polyps and 6 myoma were described. In this material the sensitivity of conventional transvaginal grey-scale ultrasound, the color Doppler ultrasound, and hysteroscopy in the detection of circumscript intrauterine lesions was 0.74, 0.93 and 1, respectively; in the diagnosis of the intrauterine lesion was 0.66, 0.91 and 0.97, respectively. Statistically significant differences were found between the ultrasound detection rates (p < 0.05) and between the diagnosis rates (p < 0.01) of the TVS and the TVCD group. Abnormal menopausal bleeding, recurrent bleeding, infertility, and tamoxifen therapy were present with high frequency in clinical data. CONCLUSION: Visualization of the feeding vessels using color or power Doppler ultrasound imaging improves the detection rate and the ultrasound diagnosis of the circumscript intrauterine lesions, approaching the sensitivity of hysteroscopy. First line application is recommended for patients at risk, such as infertility, recurrent postmenopausal bleeding, or for patients receiving long term tamoxifen.

Adult↗

Endometrial pattern and thickness associated with pregnancy outcome after assisted reproduction technologies.

The endometrial pattern and thickness was analysed prospectively on the day of administration of human chorionic gonadotrophin (HCG) in 200 in-vitro fertilization (IVF), gamete intra-Fallopian transfer (GIFT) and tubal embryo transfer (TET) cycles. Increasing maturity of the endometrial pattern was positively correlated with oestradiol levels (r = 0.20; P = 0.005), number of mature eggs (r = 0.13; P less than 0.05) and the number of top quality embryos (r = 0.40; P less than 0.001). The endometrial thickness was positively correlated with the number of follicles greater than or equal to 15 mm (r = 0.15; P less than 0.02) and the cycle day on which HCG was administered (r = 0.14; P less than 0.03). It was unaffected by the dose of human menopausal gonadotrophin and was negatively correlated with the use of clomiphene citrate (r = 0.40; P less than 0.001). Fecundity was increased for IVF when the endometrial thickness was greater than or equal to 9 mm (P less than 0.05) and for GIFT and TET when a Type C triple-line endometrial pattern was present (P less than 0.05). Biochemical pregnancies for the combined methods increased from 2.5% of all pregnancies when the endometrial thickness was 9-13 mm, to 27.8% when the thickness was less than 9 mm or greater than 13 mm (P less than 0.01). Biochemical pregnancies occurred in 67% of IVF pregnancies when the endometrial thickness was greater than or equal to 3 mm.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) inhibition of coronary development is preceded by a decrease in myocyte proliferation and an increase in cardiac apoptosis.

BACKGROUND: 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) causes cardiovascular toxicity, culminating in edema, hemorrhage, and mortality in piscine, avian, and mammalian embryos. To elucidate the mechanism of the cardiovascular teratogenicity of TCDD, we used a chick embryo model to determine whether TCDD alters coronary artery development and whether this alteration was associated with apoptosis and/or changes in myocyte proliferation. METHODS: Fertile chicken eggs were injected with corn oil (control), 0.24, or 0.40 pmol TCDD/g in corn oil before incubation. To evaluate effects of TCDD on differentiation of coronary arteries, chick embryo hearts from incubation days 8 (D8), D10, and D12 were stained with anti-alpha-smooth muscle actin. Myocyte proliferation was measured by BrdU incorporation on D6, 8, 10, and 12 after TCDD treatment. In addition, temporal and spatial patterns of apoptosis were detected by TUNEL on D3, D5, D6, D8, and D10, and immunohistochemistry was used to identify the origin of apoptotic cells on D6. RESULTS: TCDD increased apoptosis in structures where cell death normally occurs, including the outflow tract, endocardial cushion of the atrioventricular canal, and dorsal mesocardium, peaking in intensity on D6. Immunohistochemistry revealed that cells undergoing TCDD-induced apoptosis in the dorsal mesocardium were not neural or epicardial in origin. On D8 and D10 TCDD reduced myocyte proliferation. On D10, TCDD reduced coronary artery size and on D10 and D12 TCDD induced a dose-dependent decrease in coronary artery number. CONCLUSIONS: The reduction of myocyte proliferation by TCDD preceded the reduction in coronary artery number and size, suggesting that changes in coronary development may be a consequence of reduced myocyte proliferation and a thinner ventricle wall. The peak of TCDD-induced increase in apoptosis occurred even earlier in embryo development and thus may contribute to changes in myocyte proliferation, coronary development, and cardiac structural malformations; however, a cause-and-effect relationship between apoptosis and these other events has yet to be established.

Actins↗

Synapsis and meiotic recombination analyses: MLH1 focus in the XY pair as an indicator.

BACKGROUND: Anomalies in meiotic prophase I have been related to partial or total meiotic arrest. These anomalies include an abnormal synaptic process, resulting in disorders in meiotic recombination. METHODS: In the present study, we analyse primary spermatocytes from 12 infertile men (four with non-obstructive azoospermia, six with oligoastenoteratozoospermia, one with astenoteratozoospermia and one normozoospermic) and five control fertile donors using immunocytological techniques for synaptonemal complex, meiotic recombination and centromeric proteins. RESULTS: Mean numbers of MLH1 foci per cell, frequencies of cells presenting an MLH1 focus in the XY pair and percentages of cells affected by abnormal synaptic patterns (gaps and splits) are reported for each of the infertile patients and control men. A positive correlation between the frequency of cells showing a recombination focus in the XY pair and the number of autosomal recombination foci per cell is found. CONCLUSIONS: Reduced recombination in the XY pair and an increased number of cells affected by gaps may explain some idiopathic male infertility cases. The results suggest that recombination in the XY pair could be an indicator for general recombination frequency and for a successful meiotic process.

Adaptor Proteins, Signal Transducing↗

Simulation of human luteal endocrine function with granulosa lutein cell culture.

Human granulosa cells collected from in vitro fertilization have previously been cultured to provide a system to simulate the granulosa lutein cells of the corpus luteum. In most of these systems, the cultures have been relatively short term, and attempts to simulate the normal pattern of hormone production observed during the luteal phase of the cycle have not been reported. Additionally, the hormone relaxin has generally been absent from the endocrine analysis of these systems. In this report, methods were used that supported secretion of ovarian steroids and relaxin that mimics the profiles of these hormones in vivo. This system was used to observe the endocrine responses of the granulosa lutein cells to three different protocols of CG administration designed to mimic the normal luteal phase, early pregnancy, and early pregnancy followed by pregnancy loss. The normal luteal phase was simulated by a constant baseline (0.02 IU/mL) CG model to simulate a nonconceptive cycle (baseline). The second model was baseline CG until day 8 of culture, followed by daily doubling from days 9-17 to simulate an early pregnancy (rescue-plateau). CG concentrations were then held constant from days 17-20 (5.12 IU/mL). A third model (rescue-drop) was used that was identical to the early pregnancy model except that on day 17 CG was returned to baseline concentrations (0.02 IU/mL) to simulate an early pregnancy loss. Baseline CG stimulation resulted in profiles of estrogen, progesterone, and relaxin secretion in culture that were closely related to secretory profiles previously reported in serum during the nonconceptive luteal phase. The timing of appearance of relaxin secretion and later declines in steroid and relaxin secretion paralleled that observed in serum. In the CG rescue protocols, ovarian steroids rose in response to daily doubling of CG and fell when CG either plateaued or fell. Relaxin did not show an increase in response to increasing CG, but its secretion did not drop when CG concentrations plateaued or dropped. This cell culture system model mimics the profile of ovarian steroids and relaxin seen in serum during the nonconceptive luteal phase, although the relative magnitude of the hormones was not the same as seen in vivo. It was also used to investigate responses to luteal rescue protocols designed to simulate early pregnancy and pregnancy loss. This culture system may be useful to study differences in endocrine response in granulosa cells collected from different patients and to provide information of clinical relevance. This culture system provides a model to study luteal function and its response to different protocols of luteal rescue and thus may provide insight into early pregnancy and pregnancy loss.

Adult↗

Mean ages at parities: an indirect estimation.

"The objective of this paper is to describe an indirect approach for estimating the age patterns of occurrence of birth by parity. The main concern here is not of estimating the frequency of occurrence of births (in a period of time) in relation to the population (birth rate) or in relation to the number of females (fertility). Rather, the focus here is on the ages of first, second, and subsequent births, and on their shifts over time. Essentially, the approach is an extension of Hajnal's method for using proportions single to estimate singulate mean age at marriage...." The approach is illustrated using data from the 1975 Pakistan Fertility Survey and the 1990-1991 Pakistan Demographic and Health Survey. A comment by Sultan S. Hashmi is included (pp. 560-1).

Age Factors↗

[Phase-directional management of protective plantations. II. Typical protective plantation: farmland shelterbelt].

This paper is the second part of the series reports on phase-directional management of protective plantations. Based on the fundamentals of phase-directional management for protective plantations in paper I, discussions on protective maturity, management phase, and regeneration patterns and methods for shelterbelt or windbreak were conducted. Firstly, the two points of protective maturity of Populus spp. i.e., initial protective maturity age (IPMA) and terminal protective maturity age (TPMA), were determined. According to the growth pattern of shelterbelt, the IPMA was 16 years and 24 years for local poplar and hybrid poplar shelterbelts, respectively, and TPMA was the natural age for both local and hybrid poplar shelterbelts, which was determined on the basis of change pattern of porosity. Secondly, the three management phases of shelterbelts were divided quantitatively based on the analyses of discrete degrees of diameter, IPMA and TPMA. Thirdly, the corresponding measures for phase-directional management in each phase were also developed. In pre-maturity phase, i.e., the period from establishment of shelterbelt to IPMA, the measures such as soil cultivation, irrigation, fertilization, weed clearing, intercropping and branch cutting etc. should be conducted. In maturity phase, i.e., the period from IPMA to TPMA in theory, but the period from IPMA to regeneration age in practice, the techniques of managing the shelterbelt porosity around 0.25 (the best porosity value for wind protection) should be developed. In this phase, the porosity model was constructed according to the distribution pattern of base area for the poplar shelterbelt. The classification of shelterbelt trees, i.e., 4-grade classification, was established by considering the contribution of trees to the shelterbelt structure, which can provide the standard for thinning and tending of shelterbelt. Additionally, the density controlling techniques were also discussed. In the period of regeneration, i.e., from the regeneration age to the establishment of next generation shelterbelt, the techniques of regeneration such as determination of regeneration age, arrangement of regeneration patterns should be paid great emphasis. The influences of regeneration patterns and methods on regeneration period were specially discussed, and the changes of three management phases were illustrated by different regeneration patterns in the end of this paper. These results were obtained from poplar shelterbelt, but the fundamentals could be applied in any other shelterbelts.

Agriculture↗

A review of the effects of long-acting progestogen-only contraceptives on ovarian activity.

Progestogen-only contraception acts mainly by blocking cervical mucus and preventing sperm penetration through it does have a variable pattern of contraceptive effects on the endometrium and ovary. In contrast with the complete suppression of ovarian function with combined pill or injectable use, a variable degree of endocrine activity is demonstrated in women choosing a long-acting progestogen-only contraceptive. This degree of suppression of ovarian activity explains the decrease in systemic side-effects, the rapid resumption of ovulation and recovery of fertility following the discontinuation of the method. New delivery systems of progestogens, the vaginal ring and implant, offer better and more consistent contraceptive effects.

Cervix Mucus↗

Vaginal bleeding disturbances and implantable contraceptives.

Implantable contraceptives allow safe and effective fertility regulation for up to 5 years. Currently available devices release low doses of progestogens. Disruption of vaginal bleeding patterns is almost inevitable, particularly during the initial months of use. Irregular and prolonged bleeding as well as amenorrhea are common. Irregular bleeding is unpopular with most women and unacceptable to some. This review describes the vaginal bleeding disturbances induced by modern implantable contraceptives and discusses the implications of these in terms of method use and discontinuations. The cause of the irregular bleeding is not fully understood, but recent evidence suggests that an increase in endometrial vascular fragility might precipitate vessel breakdown and, hence, breakthrough bleeding. This review discusses this evidence and outline the possible mechanisms underlying breakthrough bleeding associated with implantable contraceptives. In addition, therapies for bleeding disturbances are described and their efficacy reviewed.

Contraceptive Agents, Female↗

Cadherin-1, -2 and -4 expression in the cranial ganglia and lateral line system of developing zebrafish.

Cadherins are cell adhesion molecules that play important roles in development of a variety of tissues and organs including the nervous system. In this study we analyzed expression patterns of three zebrafish classical (type I) cadherins (cadherin-1, -2, and -4) in the embryonic zebrafish cranial ganglia and lateral line system using in situ hybridization and immunohistochemical methods. All three cadherins exhibit distinct spatiotemporal patterns of expression during cranial ganglia and lateral line system development. cadherin-1 message was detected in the trigeminal and facial ganglia, in the lateral line ganglia, and in most of neuromasts in the lateral lines. Cadherin-2 mRNA and protein were expressed by the majority of the cranial ganglia and lateral line system. Both cadherins were found in embryos younger than 24 hours post fertilization as well as in 2-3-day old embryos and larvae. In contrast, cadherin-4 mRNA and protein expression was detected in embryos older than 30 hours post fertilization and limited to the trigeminal, statoacoustic, and vagal cranial ganglia, and the lateral line ganglia of older embryos and larvae.

Animals↗

Analysis of semen in a constantly changing social context of medicine.

Semen analysis is one of the most valuable diagnostic methods that play a critical role in andrology. The introduction of computer-assisted methods of human semen analysis [6], in vitro fertilization, and other auxiliary methodologies [4] has greatly enhanced the expectations of infertile couples in having a baby. However, routine semen analysis plays a significant role in an infertility clinic, particularly in developing countries. Standardization of routine semen analysis therefore becomes the key issue in laboratories involved in such analysis. Apart from that, semen analyses carried out in different populations bring out the changing pattern of semen quality due possibly to the influences of factors like lifestyle, race, population dynamics, and culture. It also helps to set standards of semen quality in the changing context of medicine.

Humans↗

Contraception with NORPLANT implants.

NORPLANT progestogen-only implant contraception provides several years of effective protection against pregnancy following a single application. It is reversible whenever desired, with return to normal fertility. Drug release and concentrations are maximal in the first month of use, when a set of six capsules releases approximately 85 micrograms/day of levonorgestrel. Release is 25-30 micrograms/day at 60 months. Circulating drug concentrations and pregnancy rates vary inversely with body weight at placement, but even among heavier women, failure rates are below those of oral contraceptives in the general population. Side-effects are most marked at the initiation of use. Disruption of normal menstrual patterns occurs in 70-80% of recipients initially. Menstrual blood loss is decreased, however, and most women experience modest increases in haemoglobin. Headache, acne, other skin or hair problems, changes of weight and of mood and abdominal pain, are the most common side-effects attributed to NORPLANT contraception. The frequency of occurrence of side-effects diminishes with increased duration of use. The comparative paucity of major side-effects, coupled with the method's convenience and effectiveness and its very long action have made NORPLANT implants highly acceptable in both developing and developed countries.

Contraceptive Agents, Female↗

Effects of hormone replacement therapy on growth hormone secretion patterns in correlation to somatometric parameters in healthy postmenopausal women.

OBJECTIVES: The aim of the present study was to investigate the influence of a continuous estrogen, cyclic progesterone replacement therapy on the secretion of growth hormone (GH) and IGF I as well as of somatometric-GH correlation patterns. METHODS: The study included 23 healthy postmenopausal women. Of the proband group 13 randomly selected women were treated with orally applicated 2 mg estradiol-valerat (E2V) and 10 mg dydrogesterone for 10 months. Ten women did not receive any hormonal treatment during this time. After 10 months all probands were reexamined and their GH and IGF I secretion, as well as their somatometric-hormonal correlation patterns, compared with those of a fertile control group. RESULTS: It could be shown, that in postmenopausal women a 10-month oral hormone replacement therapy led to a significant increase of GH- and IGF I levels, however, the treated postmenopausal women did not reach the levels of the fertile controls. Those women who did not receive any hormonal treatment and the postmenopausal women before HRT showed nearly identical GH- and IGF I levels as well as somatometric-GH correlation patterns. CONCLUSIONS: The results of the present paper indicate a marked influence of estrogens on GH and IGF I secretion. Furthermore, hormonal replacement therapy (HRT) may influence somatometric GH correlation patterns too.

Adult↗

Homopolymeric tract heteroplasmy in mtDNA from tissues and single oocytes: support for a genetic bottleneck.

While mtDNA polymorphisms at single base positions are common, the overwhelming majority of the mitochondrial genomes within a single individual are usually identical. When there is a point-mutation difference between a mother and her offspring, there may be a complete switching of mtDNA type within a single generation. It is generally assumed that there is a genetic bottleneck whereby a single or small number of founder mtDNA(s) populate the organism, but it is not known at which stages the restriction/amplification of mtDNA subtype(s) occur, and this uncertainty impedes antenatal diagnosis for mtDNA disorders. Length polymorphisms in homopolymeric tracts have been demonstrated in the large noncoding region of mtDNA. We have developed a new method, T-PCR (trimmed PCR), to quantitate heteroplasmy for two of these tracts (D310 and D16189). D310 variation is sufficient to indicate clonal origins of tissues and single oocytes. Tissues from normal individuals often possessed more than one length variant (heteroplasmy). However, there was no difference in the pattern of the length variants between somatic tissues in any control individual when bulk samples were taken. Oocytes from normal women undergoing in vitro fertilization were frequently heteroplasmic for length variants, and in two cases the modal length of the D310 tract differed in individual oocytes from the same woman. These data suggest that a restriction/amplification event, which we attribute to clonal expansion of founder mtDNA(s), has occurred by the time oocytes are mature, although further segregation may occur at a later stage. In contrast to controls, the length distribution of the D310 tract varied between tissues in a patient with heteroplasmic mtDNA rearrangements, suggesting that these mutants influence segregation. These findings have important implications for the genetic counselling of patients with pathogenic mtDNA mutations.

Animals↗

Temporal expression profiling of the uterine luminal epithelium of the pseudo-pregnant mouse suggests receptivity to the fertilized egg is associated with complex transcriptional changes.

BACKGROUND: The molecular basis of changes underlying the altered sensitivity of the uterine luminal epithelium (LE) to the embryo over the peri-implantation period is not fully understood. METHODS: Microarray analysis was performed on purified LE isolated from the pseudo-pregnant mouse uterus at 12-h intervals from pre-receptivity through the implantation window to refractoriness. The aim was to identify genes whose expression changes in the LE during this period. RESULTS: A total of 447 transcripts were identified whose abundance changed more than 2-fold in the LE but which did not change in the underlying stroma (S) and glands. Six major patterns of changing expression were noted. Of the 447 genes, 140 were expressed in LE at least 15-fold higher than in S and glandular epithelium (GE) (101 of these more than 20-fold). Detailed spatiotemporal expression profiles were derived for several genes previously implicated in implantation (including Edg7, Ptgs1, Pla2g4a and Alox15). CONCLUSIONS: Functional changes in LE receptivity are characterized by changing constellations of gene expression. Pre-receptivity has a different molecular footprint to refractoriness. Because we have used the pseudo-pregnant mouse model, these changes are driven solely by endocrine signals rather than events downstream of embryo attachment. Some of these genes have been described in previous microarray studies on endometrium, but for the majority, this is the first time they have been implicated in implantation. The 140 genes enriched in the LE greatly expand the list of epithelial markers and provide many novel candidates for further studies to identify genes playing important roles in receptivity and embryo attachment.

Animals↗

Development of embryos from in vitro ovulated and fertilized oocytes of the quail (Coturnix coturnix japonica).

The development of quail embryos obtained after in vitro fertilization of oocytes ovulated in vitro was investigated. About 40% of the specimens, after 18-20 hr of incubation, had undergone cleavage to reach stages IV-VI when viewed under a stereo microscope. However, only 36% of these embryos contained normal, DAPI-stained nuclei when observed under a fluorescent microscope; the other 64% showing a morphologically normal cleavage pattern did not contain nuclei. Control unfertilized oocytes, ovulated in vitro and cultured for the same time, also sometimes attained the morphologically correct stages IV-VI but their "blastomeres" were always devoid of nuclei. Therefore, it is advisable to monitor early avian embryos for the presence of nuclei when assessing development in culture. The results demonstrate, for the first time, that cytoplasmic segmentation can occur in the absence of nuclear divisions in the germinal disc of the quail and show the existence and significance of ooplasmic maternal information in birds. This phenomenon is also known for sea urchin and frogs. It is indicative of the role of maternal information in early development. The in vitro method presented here links the steps of ovulation and fertilization with the early cleavage stages under in vitro conditions and may be useful in studying mechanisms of fertilization and differentiation in birds as well as in obtaining transgenic birds by DNA injection or application of foreign, DNA-carrying sperm.

Animals↗

Identification of lymphocyte subsets in the human fallopian tube.

PROBLEM: Immunohistochemical investigations for the detection of lymphocyte subsets in the human oviduct have been performed. Knowledge about local immunity especially cell-mediated immunity, in the fallopian tube has been, up to now, limited. As an essential structure for the human reproduction process, the tubal mucous membrane is exposed to a variety of antigens. METHOD: A total number of 20 tubal biopsies obtained from fertile women during gynecological operations like tubal ligations or hysterectomy were examined by the immunoperoxidase technique. Seven specimens were obtained during the proliferative phase, ten during the secretory phase and three during a caesarean section with tubal ligations. RESULTS: It could be established that the presence of lymphocytes in the oviductal mucous membrane is physiological. These cells can be identified by their typical immunohistochemical patterns. There were no significant differences of the type and number of lymphocytes in the mucosa within the phases of menstrual cycle. The dominant cell types in the tubal mucosa were the CD3+ and CD8+ lymphocytes. CONCLUSIONS: It can be suggested that the lymphocytes in the tubal mucosa may involved in the process of immune tolerance, which could realize the transport of sperms and blastocysts through the oviduct under normal conditions without activation of local immune mechanisms. The lymphoid tissue of the oviduct is a specialized form of mucosal-associated lymphoid tissue (MALT).

Adult↗