Association of maternal angiotensin II type 1 and type 2 receptor combination genotypes with susceptibility to early-onset preeclampsia.

Allelic variations affecting the activity of the maternal renin-angiotensin system may play a role in the development of hypertensive disorders of pregnancy like preeclampsia, its more severe early-onset form, and intrauterine growth restriction. We examined the association of common allelic variants of angiotensin II type 1 and type 2 receptor genes (AT1R and AT2R) sorted in five AT1R/AT2R receptor combination genotype groups with susceptibility to early-onset preeclampsia (EOP). The occurrence of AT1R (A1166C) and A2TR (C3123A) alleles in wild type (AA, CC), heterozygous (A/C, C/A), and homozygous (C/C, A/A) states was recorded in 84 women with a history of EOP and 84 age-matched controls sorted in five AT1R/AT2R receptor combination genotype (wild type: AA/CC, one mutant: AA/CA, AC/CC, two mutant: AC/CA, AA/AA, CC/CC, three mutants: AC/AA, CC/CA and four mutant: CC/AA) groups, by polymerase chain reaction-RFLP analysis. Three mutant receptor combination genotype carriers were more common in women with a history of EOP than in controls (26.18% vs. 4.76%, p = 0.003, OR = 8.25). Receptor combination genotyping may be of clinical value in: (a) maternal prediction of susceptibility to EOP, (b) disease subtyping for directed studies with receptor signaling antagonists, (c) the broader study of hypertension.

Chorionic villi derived mesenchymal like stem cells and expression of embryonic stem cells markers during long-term culturing.

Mesenchymal stem cells (MSCs) can be obtained from a variety of human tissues. MSCs derived from placental chorionic villi of the first trimester are likely to resemble, biologically, embryonic stem cells (ESC), due to the earlier development stage of placenta. In the present study long-term cultures of MSC-like cells were assessed in order to evaluate MSCs multipotent characteristics and molecular features during the period of culture. CV-cells obtained from 10 samples of chorionic villus displayed typical fibroblastoid morphology, undergone 20 passages during a period of 120 days, maintaining a stable karyotype throughout long term expansion. The cells were positive, for CD90, CD73, CD105, CD29, CD44, HLA ABC antigens and negative for CD14, CD34, AC133, and HLA DR antigens as resulted from the flow cytometry analysis. CV-cells were differentiated in adipocytes, osteoblasts, chondrocytes and neuronal cells under specific culture conditions. The expression of the ESC-gene markers POU5F1 (Oct-4) and NANOG was observed at earliest stages (4-12 passages) and not at the late stages (14-20 passages) by RT-PCR analysis. ZFP42 and SOX2 expression were not detected. Moreover, CV-cells were found to express GATA4 but not NES (Nestin). Chorionic villi-derived cells possess multipotent properties, display high proliferation rate and self-renew capacity, share common surface antigens with adult MSCs and express certain embryonics stem cells gene markers. These characteristics highlight chorionic villi as an attractive source of MSCs for the needs of regenerative medicine.

First trimester diagnosis of 13q-syndrome associated with increased fetal nuchal translucency thickness. Clinical findings and systematic review.

13q-syndrome is a rare chromosomal disorder caused by partial deletion of the long arm of chromosome 13 with variable phenotypic presentation. Further sonographic features involve fetal growth restriction, bradycardia, encephalocele, facial dysmorphism and upper extremity deformity. We report a case of 13q-syndrome presenting as increased nuchal translucency diagnosed by chromosome studies and confirmed by array comparative genomic hybridization (CGH) analysis in the first trimester of pregnancy. Pregnancy was terminated at 14 weeks' gestation. The parents did not give consent for a postmortem examination. Furthermore we performed a systematic review of the international literature on previous cases of 13q-syndrome diagnosed prenatally. Our case emphasizes the importance of a detailed 11-14 week ultrasound assessment in diagnosing fetal chromosomal aberrations in combination with the modern aspects of array CGH, thus providing more precise and rapid prenatal diagnosis.

Diagnostic accuracy of nerve fibre layer, macular thickness and optic disc measurements made with the RTVue-100 optical coherence tomograph to detect glaucoma.

PURPOSE: To evaluate the diagnostic accuracy of retinal nerve fibre layer thickness (RNFLT), ganglion cell complex (GCC), and optic disc measurements made with the RTVue-100 Fourier-domain optical coherence tomography (OCT) to detect glaucoma in a Caucasian referral population. METHODS: One randomly selected eye of 286 Caucasian patients (93 healthy, 36 ocular hypertensive, 46 preperimetric glaucoma, and 111 perimetric glaucoma eyes) was evaluated. RESULTS: Using the software-provided classification, for the total population sensitivity did not exceed 73.6% for the optic nerve head parameters, and 62.7% for the other parameters. Specificity was high (94.6-100%) for most RNFLT and GCC parameters, but low (72.0-76.3%) for the optic disc parameters. Positive predictive value varied between 98.1 and 100% for the main RNFLT parameters, 92.6 and 100% for the 16 RNFLT sectors, 92.4 and 99.0% for the GCC parameters, but did not exceed 86.3% for any of the optic disc parameters. Positive likelihood ratio (PLR) was higher than 10 for average, inferior and superior RNFLT (25.5 to infinite), 12 of the 16 RNFLT sectors (12.6 to infinite), and three of the four GCC parameters (40.0 to 48.6). No optic disc parameter had a PLR higher than 3.0. CONCLUSIONS: RNFLT and GCC parameters of the RTVue-100 Fourier-domain OCT showed moderate sensitive but high specificity, positive predictive value and PLR for detection of glaucoma. The optic disc parameters had lower diagnostic accuracy than the RNFLT and GCC parameters.

Characterization of 23 small supernumerary marker chromosomes detected at pre-natal diagnosis: The value of fluorescence in situ hybridization.

Small supernumerary marker chromosomes (sSMCs) cannot be identified or characterized unambiguously by conventional cytogenetic banding techniques. Until recently, the large variety of marker chromosomes, as well as the limitations in their identification, have presented a diagnostic problem. In order to determine the origin of sSMCs, we used a variety of fluorescence in situ hybridization (FISH) methods, including centromere-specific multicolor FISH, acrocentric specific multicolor FISH, subcentromere-specific multicolor FISH and multicolor FISH with whole chromosome paint probes. Moreover, uniparental disomy testing was in all cases attempted. From a total of 28,000 pre-natal samples from four diagnostic genetics laboratories in Greece, 23 (0.082%) supernumerary marker chromosomes were detected. The mean maternal age was 36.2 years (range 27-43) and the mean gestational age at which amniocentesis was performed was 18.5 weeks (range 16-23). Eighteen markers were de novo and 5 markers were inherited. Molecular cytogenetic methods were applied to determine the chromosomal origin and composition of the sSMC. In total, 17 markers were derived from acrocentric chromosomes (14, 15, 21 and 22) and 6 markers were non-acrocentric, derived from chromosomes 9, 16, 18, 20 and Y. Uniparental disomy was not detected in any of the cases studied. With regard to pregnancy outcome, 13 pregnancies resulted in normal healthy neonates, while 10 pregnancies were terminated due to ultrasound abnormalities. A total of 23 marker chromosomes from 28,000 pre-natal samples (0.082%) were identified. Molecular cytogenetic techniques provided valuable information on the chromosomal origin and composition of all the sSMCs. Especially in cases with normal ultrasound, the FISH results rendered genetic counseling possible in a category of cases previously considered a diagnostic problem. Abnormal outcome was observed in 10 cases (43,5%), 7 of which showed abnormal ultrasound findings. New technologies, such as array-comparative genomic hybridization, should be used in future genotype-phenotype correlation studies, although the high mosaicism rate poses a problem.

The effect of diclofenac and paracetamol on pregnancy and implantation rates in infertile women undergoing IVF treatment.

We set out to assess the effect of diclofenac on implantation rates, when administered as analgesia following transvaginal oocyte recovery. This was a prospective study of infertile women undergoing IVF treatment in a University Hospital. Subjects (n = 74) were divided in two groups (A and B). Group A (n = 38) received 1 g paracetamol and 100 mg diclofenac and group B (n = 36) received 1 g paracetamol only. All the medication was administered rectally immediately after the oocyte retrieval. Pregnancy and implantation rates were compared between group A and B using the chi2 test. In groups A and B, the implantation rates were 12.4% and 9.6% (p = 0.5) and the pregnancy rates were 28.9% and 19.4%, respectively (p = 0.67). Neither pregnancy nor implantation rates differed significantly between the two groups. Administration of diclofenac to patients at the time of egg collection does not appear to affect implantation or pregnancy rates, while it could be effective in reducing discomfort and pain associated with oocyte retrieval.

Oestradiol stimulates prolactin secretion in women through oestrogen receptors.

OBJECTIVE: To examine the effects of clomiphene and raloxifene on basal and GnRH-induced prolactin (PRL) secretion in postmenopausal women. DESIGN: Postmenopausal women participated in two experimental procedures a month apart. In one experiment they received raloxifene (180 mg/day) (R-Exp) and in the other clomiphene (150 mg/day) (Cl-Exp). In Group 1, the women (n = 8) received raloxifene or clomiphene for 30 days plus oestradiol via skin patches (100 microg/24 h) for the last 10 days. In Group 2, the women (n = 8) received oestradiol for 30 days plus raloxifene (R-Exp) or clomiphene (Cl-Exp) for the last 10 days. The pituitary response to GnRH (100 microg i.v.) was investigated in all women on days 0, 10, 20 and 30 of each experiment. PATIENTS: The study included 16 healthy postmenopausal volunteer women aged 56-60 years. MEASUREMENTS: Basal levels of PRL and the area under the curve (AUC) of DeltaPRL response to GnRH were calculated. RESULTS: In Group 1, basal levels of PRL and the area under the curve (AUC) of PRL response to GnRH did not change significantly in both experiments. In Group 2, during both experiments basal levels of PRL and the AUC of PRL increased significantly on days 10 (P < 0.05) and 20 (P < 0.05) as compared to day 0 and then they decreased significantly on day 30 as compared to day 20 (P < 0.05). CONCLUSIONS: Our study demonstrates for the first time that raloxifene and clomiphene affect the secretion of PRL in postmenopausal women in a similar manner. It is suggested that oestradiol stimulates the secretion of PRL in women by acting through oestrogen receptors.

Effects of clomiphene and raloxifene on gonadotrophin secretion in postmenopausal women: evidence of nongenomic action.

OBJECTIVE: To evaluate the effect of raloxifene (R) and clomiphene (Cl) on FSH and LH secretion in postmenopausal women. DESIGN: Postmenopausal volunteer women participated in two experimental (Exp) procedures. In Group 1, the women received R (180 mg/day orally) for 30 days plus oestradiol (E2) through skin patches (100 microg/24 h) from days 21 to 30 (R-Exp). After a month's break the same women received Cl (150 mg/day orally) for 30 days plus E2 as above (Cl-Exp). In Group 2, the women received E2 for 30 days plus R from days 21 to 30 (R-Exp) and after a month's break they received E2 for 30 days plus Cl from days 21 to 30 (Cl-Exp). Daily doses were as in Group 1. A GnRH test (100 microg intravenously) was performed in all women on days 0, 10, 20 and 30 of each experiment. PATIENTS: Sixteen healthy postmenopausal women were divided into two groups (eight women in each group). MEASUREMENTS: The area under the curve (AUC) of DeltaFSH and DeltaLH response to GnRH (net increase above the basal value) was calculated. RESULTS: In Group 1, basal levels of FSH and LH did not change significantly during the R-Exp, while they decreased significantly in the Cl-Exp (P < 0.001). The addition of E2 did not have any effect. The AUC of LH response to GnRH increased significantly in the R-Exp (P < 0.05) and that of FSH in the Cl-Exp (P < 0.05). In Group 2, basal levels of FSH and LH declined significantly during treatment with E2 in both the R-Exp (P < 0.01) and the Cl-Exp (P < 0.001). However, the addition of Cl (for 10 days) interrupted this decrease, while the addition of R stimulated FSH levels significantly (P < 0.05). E2 suppressed significantly the AUC of LH in both experiments (P < 0.05). The addition of Cl did not affect the AUC in response to GnRH, while the addition of R increased the AUC of both LH and FSH (P < 0.05). CONCLUSIONS: These results demonstrate for the first time that in contrast to Cl, R does not exert oestrogenic effects on basal gonadotrophin secretion. Although the antioestrogenic action of these drugs was evident only after pretreatment with E2, both R and Cl stimulated GnRH-induced gonadotrophin secretion in oestrogen-deprived women. It is hypothesized that these two compounds sensitize the pituitary to GnRH through mechanisms not involving the oestrogen receptor complex (nongenomic).

Neonatal magnetocardiography and Fourier spectral analysis.

OBJECTIVE: In this study we investigated the relationship between heart rate variability (HRV) and respiration in healthy and probably asphyxiated infants using magnetocardiographic (MCG) recordings. METHODS: Ten healthy and eight infants from pregnancies complicated by preeclampsia with indications of mild perinatal asphyxia were included in the study. All were near term. Maternal age ranged from 16 to 39 years (mean = 29.05, SD = 6.13). Spectral analysis was used to find out any association between respiration and HRV. RESULTS: Respiratory sinus arrhythmia (RSA) was reduced in preeclamptic infants with indications of mild perinatal asphyxia. This difference was statistically significant (p < 0.0002, t-test), whereas the heart rate of the two groups was not statistically significant (p = 0.1, t-test). CONCLUSION: The results suggest that infants with indications of mild preeclampsia differ from controls in respiratory activity and this difference is independent of basal heart rate. Thus, spectral analysis could be useful for the estimation of influence of mild perinatal asphyxia in the RSA rhythm of newborns.

Neonatal magnetocardiography.

PURPOSE: The aim of the present study was to test the validity of magnetocardiography (MCG) in the estimation of neonatal cardiac rhythm using a single channel superconductive quantum interference device (SQUID). MATERIALS AND METHODS: Our study population consisted of 50 neonates who were delivered normally between 37-41 weeks of gestation from clinically uncomplicated pregnancies. There was also a neonate included in the study in which the diagnosis of "hypoplastic left heart syndrome" was demonstrated by U/S Doppler examination. Maternal age ranged from 18 to 39 years (mean=29.15, SD=6.13). RESULTS: Our study results revealed 44 neonates with normal cardiac rhythm, four with ventricular tachycardia (VT), one with ventricular tachycardia (VT) and extrasystolic beats and one with bradycardia. The neonate with the hypoplastic left heart syndrome presented frequent episodes of ventricular bigeminy in the magnetocardiographic trace. M-mode echocardiography confirmed the diagnosis of the seven cases of arrhythmia in our study group. CONCLUSION: Results gained from the study lead us to believe that MCG could provide clinical practice with a non-invasive, rapid and easy to perform method, which could be used as an adjunct to conventional methods for the evaluation of neonatal cardiac rhythm.

Three-dimensional kinematic analysis of the snatch of elite Greek weightlifters.

We investigated the linear kinematics and the change in energy of the barbell and the angular kinematics of the trunk and leg during the snatch technique of 12 elite male Greek weightlifters under competitive conditions after the new weight classification. Two S-VHS cameras operating at 60 Hz were used to record the lifts. The spatial coordinates of selected points were calculated using the direct linear transformation procedure; after digital filtering of the raw data, the angular displacements and angular velocities were calculated for the hip, knee and ankle joints. The following variables were also calculated for the barbell: vertical and horizontal displacement, vertical linear velocity and acceleration, external mechanical work and power output. The results revealed that all weightlifters flexed their knees during the transition phase, independently of their weight category. This indicates that the athletes use the elastic energy produced during the stretch-shortening cycle to enhance their performance. In nine athletes, we found that the barbell trajectory did not cross a vertical reference line that passed through the initial position of the barbell. The vertical linear velocity of the barbell was increased continuously from the beginning of the movement until the second maximum extension of the knee joint, with no notable dip being observed. Regarding the change in energy of the barbell, we found that the mechanical work for the vertical displacement of the barbell in the first pull was significantly greater than the mechanical work in the second pull. In contrast, the estimated average mechanical power output of the athletes during the vertical displacement of the barbell was significantly greater in the second pull than in the first pull. We conclude that the major elements of the snatch technique of elite Greek weightlifters have not been affected by the new weight classification.