bluuebunny/crossref_metadata_2025_split huggingface dataset:
ORIGINAL ARTICLE: An Efficient Optimized Method for Isolation of Villous Trophoblast Cells from Human Early Pregnancy Placenta Suitable for Functional and Molecular Studies
Authors: Stenqvist Ann‐Christin , Chen Ting , Hedlund Malin , Dimova Tanya , Nagaeva Olga , Kjellberg Lennart , Innala Eva , Mincheva‐Nilsson Lucia 
Published: June 2008

Problem  The uniqueness of the human placenta cannot be replaced by animal models. In vitro studies are compulsory to elucidate the biology of human placenta and require isolation and purification of villous trophoblasts, which can be used in molecular and functional studies. Constant improvement in the isolation technique is required to obtain a high yield of pure trophoblast cells with high viability and well preserved morphology.Method of study  Optimized isolation procedure for human villous trophoblasts based on mild enzymatic treatment, Percoll gradient centrifugation and additional purification step involving positive or negative immunoselection on magnetic beads is described.Results  A simple and effective isolation protocol gave a reasonably high yield of villous trophoblast cells with high purity and viability, and excellent morphology as assessed by flow cytometry and electron microscopy.Conclusion  This protocol provides an efficient, optimized method for isolation and enrichment of villous trophoblast cells, suitable for phenotypic, ultrastructural, molecular and functional analyses and for establishment of primary cultures.
ORIGINAL ARTICLE: Immunophenotype of Peripheral T Lymphocytes, NK Cells and Expression of CD69 Activation Marker in Patients with Recurrent Spontaneous Abortions, During the Mid‐Luteal Phase
Authors: Prado‐Drayer Aramilena , Teppa José , Sánchez Pedro , Camejo María Isabel 
Published: June 2008

Problem  Determination of subpopulations of T lymphocytes, natural killer (NK) and activation status, in peripheral blood during the mid‐luteal phase from patients with unexplained recurrent spontaneous abortion (RSA).Method of study  Peripheral blood samples from non‐pregnant women with RSA and normal multiparous were taken and evaluated for subpopulations of T lymphocytes: CD4, CD8, (‘naive‐like’ and ‘memory‐like’), TCR receptor (αβ and γδ), activation status by CD69+surface or intracellular/CD3+, and NK cells (CD16+/CD56dim/CD3−, CD16+/CD56 bright/CD3−, CD69+surface or intracellular/CD56+/CD3− cells).Results  The evaluation of T lymphocytes only showed an increase in the expression of CD69 (surface and intracellular) in the RSA group. Additionally, we observed an increase in the total NK cells, CD56+ NK cells percentages, CD56dim NK cells and CD69 NK cells in RSA group.Conclusion  These observations support the concept that immunological activation of T lymphocytes and NK cells could be involved in peripheral blood during the mid‐luteal phase in patients with unexplained RSA.
ABSTRACTS: 10 Low doses of trilostane fail to induce abortion during early gestation in mice and ewes
Authors: Solano ME , Parker V , Camous S , Sandra O , Douglas AJ , Arck PC 
Published: June 2008

Problem:  Progesterone plays a central role during the maintenance of pregnancy. Our study aims to develop two hypoprogesteronic animal models using the 3beta‐HSD inhibitor trilostane during early pregnancy.Material and Methods:  Trilostane or vehicle was injected s.c. in DBA/2J‐CBA/J female mice (4.5‐8.5 dpc) or in ewes (5‐16 day of pregnancy). Progesterone serum levels were assayed (mouse: 13.5 dpc; ewe: day 16). The resorption rate (RR), number of implantation sites (IS, mouse) or development of the conceptus (sheep) were analyzed.Results:  Between trilostane‐treated or control females progesterone levels did not differ in the mouse but, as expected, progesterone did not rise in the sheep model. Nevertheless, ovine conceptus development was not morphologically affected nor IS or RR in the mouse model.Conclusions:  our data suggest that trilostane dose was insufficient to affect the reproductive success in our animal models. A more thorough analysis is currently undertaken in order to better characterize the phenotypes.
ABSTRACTS: 11 Effect of an immune stress on prolactin secretion in early pregnancy
Authors: Parker VJ , Douglas AJ 
Published: June 2008

Problem:  Stress in early pregnancy has been linked to increased abortion rates. Immune stress in early pregnancy inhibits progesterone secretion in mice. As prolactin both mediates implantation and drives progesterone secretion, we hypothesised that stress would decrease prolactin secretion in early pregnancy.Material and Methods:  Lipopolysaccharide (LPS; 12.5μg intraperitoneally) as an immune stress or vehicle were administered to day 5.5 early pregnant and virgin c57/Bl6J mice. They were killed by decapitation 60, 120 or 240 min later and trunk blood was collected and analysed for prolactin concentration (ELISA); corticosterone was also analysed (RIA).Results:  LPS significantly decreased prolactin concentration in early pregnancy (P<0.001, 2‐way ANOVA); however, there was no significant difference in the virgin groups. In contrast, LPS significantly elevated corticosterone concentration in all groups (P<0.001, 2‐way ANOVA), confirming activation of the hypothalamo‐pituitary‐adrenal stress axis.Conclusions:  Stress decreased prolactin secretion during early pregnancy, and might explain stress‐disrupted implantation and progesterone secretion.
ABSTRACTS: 12 The complement component C1q: A novel angiogenic factor?
Authors: Bossi Fleur , Rizzi Lucia , Munot Carine , Berton Stefania , Lorenzetto Erica , Baldassarre Gustavo , Frankenne Francis , Tedesco Francesco 
Published: June 2008

Aim:  Decidual endothelial cells are able to synthesize C1q and express surface‐bound C1q under physiological conditions. Since decidua is a site of active angiogenesis, we sought to ascertain whether C1q could play a role in this process.Material and Methods:  To confirm our hypothesis we used different approaches such as permeability, cell migration and proliferation assay, besides wound healing and aortic ring assay.Results:  C1q acts as a permeabilizing factor inducing the FITC‐BSA leakage through a monolayer of endothelial cells (ECs). Next, we found that C1q was able to promote motility of ECs in a wound healing assay, and to recruit ECs acting as a chemotactic factor, furthermore C1q was also found to have an additional effect on EC inducing cell proliferation. To confirm and extend these data, we used the rat aortic ring assay to evaluate the ex vivo effect of C1q.Conclusion:  Based on our findings, we propose that C1q exerts unexpected proangiogenic effects.
ABSTRACTS: 2 Mouse PAI‐1 promotes placentation by increasing foetal and maternal angiogenesis
Authors: Labied S , Munaut C , Blacher S , Coqué N , Sandra O , Noël A , Carmeliet P , Foidart J‐M , Frankenne F 
Published: June 2008

Introduction:  Murine placentation is associated with trophoblast cells invasion of the maternal endometrium and extensive maternal and foetal angiogenesis. Both processes involve proteases‐dependent extracellular matrix remodelling. Among the protease inhibitors, plasminogen activator inhibitor‐1 (PAI‐1) is transiently produced by spongiotrophoblasts and trophoblast giant cells at days 10.5‐11.5 day post‐coitum (dpc). Although accumulating evidence demonstrates the key role of PA‐1 in pathological angiogenesis, its function during placental vascularisation remains to be elucidated. PAI‐1 knockout mice are fertile and the litter sizes are normal. We have therefore analysed the consequence of PAI‐1 deficiency on murine placentation.Material and Methods:  We have studied the possible role of PAI‐1 by quantitating the placental vessel density, the relative thickness of the labyrinth, decidua and spongiotrophoblast at day 10.5, 12.5 and 14.5 dpc in mice deficient for PAI‐1 or in control mice. An original method of computer‐assisted image analysis allowed us to quantify alterations of several placental compartments identified with specific monoclonal antibodies (keratin, desmin, fibrinogen and MECA‐32). To investigate the differentially expressed genes, we performed laser capture microdissection (LCM), followed by genome‐wide expression profiling using high‐density oligonucleotides microarray analysis (GeneChip Mouse Genome 430 2.0 Array, Affymetrix). Data were analysed using Ingenuity Pathways Analysis (Ingenuity Systems®, http://www.ingenuity.com).Results:  At 10.5 and 12.5 dpc, an abnormal placental morphology was observed in both labyrinth and spongiotrophoblast layers in PAI‐1‐/‐ mice. Lack of PAI‐1 resulted in a transient decreased maternal and fetal vascularisation of the placenta that caused (1) an enhancement in the decidua/labyrinth and labyrinth/spongiotrophoblast thickness ratios, (2) a significant increase of trophoblast density. Normalization of placental morphology occurred by day 14.5 dpc in PAI‐1 deficient mice. Statistical analysis of microarrays revealed 706 genes differentially expressed between PAI‐1 deficient and normal mice in the labyrinth zone at 10.5 dpc. At 14.5 dpc, only 205 genes are differentially expressed. Using Ingenuity Pathways Analysis, most of those genes were found to be associated to lipid metabolism, cellular growth and proliferation.Conclusion:  Despite a transient PAI‐1 requirement for optimal placental angiogenesis, this gene does not appear to be essential for trophoblast invasion and placentation.
ABSTRACTS: 3 Development of an in vitro model for human embryo implantation and progesterone regulation
Authors: Lalitkumar Sujata , Lalitkumar PGL , Meng ChunXia , Gemzell‐Danielsson Kristina 
Published: June 2008

Aim:  Develop an in vitro 3D‐model for human embryo implantation on receptive endometrial construct.Material and Methods:  Three‐dimensional in vitro endometrial construct (n=12) was developed by culturing isolated human endometrial stromal cells in collagen matrix overlaid with epithelial cells. After priming hormonally, human blastocysts were placed and cultured in alpha‐medium. Attachment of embryo was confirmed using cytokeratin‐7 and expression of endometrial receptivity markers using immunohistochemistry. Antiprogestin mifepristone was used to test implantation and receptivity.Results:  Embryos attached to receptive endometrial construct. Epithelial and stromal cells of endometrial construct expressed ER‐α, ER‐β, PR‐(A+B), PR‐B, VEGF, LIF, IL‐1beta and COX‐2. Epithelial cells expressed AR, integrinαVβ3 and MUC1. Mifepristone inhibited blastocyst attachment, up‐regulated epithelial ER‐β, PR‐B; down regulated stromal VEGF, MUC1 and integrin αVβ3 as observed in vivo.Conclusion:  An in vitro human embryo implantation model, regulated by progesterone was developed and tested. This could be further used to understand molecular basis of human embryo implantation.
ABSTRACTS: 4 Modulation of maturation and function of dendritic cells by female sex steroid hormones
Authors: Segerer SE , Müller N , Van Den Brandt J , Kapp M , Dietl J , Reichardt HM , Rieger L , Kämmerer U 
Published: June 2008

Problem:  Pregnancy represents an exclusive situation in which the immune and the endocrine system cooperate to prevent rejection of the embryo by the maternal immune system. While immature dendritic cells (iDC) in the early pregnancy decidua presumably contribute to the establishment of peripheral tolerance, hormones like estradiol (E2), progesterone (Prog) and bHCG are candidates that could direct the differentiation of DCs into a tolerance‐inducing phenotype.Methods and Results:  To test this hypothesis we generated iDCs from peripheral‐blood‐monocytes and exposed them to E2, Prog, bHCG and Dexamethasone (Dex) as control. Surprisingly, E2, Prog and bHCG upregulated the expression of HLA‐DR, CD 40, CD 83 and CD 86. Visualization of the F‐actin cytoskeleton confirmed these observations. In contrast, the T‐cell stimulatory capacity of DCs was reduced after E2, Prog and bHCG exposure.Conclusion:  These findings suggest that E2, Prog and bHCG interfere with selected aspects of DC maturation and may thereby help preventing activation of allogenic T‐cells by the embryo.
ABSTRACTS: 5 In vivo gene delivery of the murine uterus: what and why?
Authors: Rodde N , Munaut C , Prince S , Olivier F , Bellemin AL , Chaouat G , Sandra O 
Published: June 2008

Problem:  The in vivo gene delivery offers an alternative for affecting the expression of genes without deriving transgenic animals. In order to investigate the biological functions of genes during implantation, the in vivo transfection of the murine uterus was assessed using a non‐cationic agent.Material and Methods:  Various routes of administration were tested using complexes of JetPEI (Polyplus‐Transfection) and pEGFPluc plasmid injected into mice. The luciferase activity was quantified and the GFP cell localisation was investigated in the uterus. The consequences of the injections on the pregnancy were estimated.Results:  The efficiency of the in vivo transfection in the uterus as well the type of transfected cells depends on the route of injection. No apparent effect was seen on the pregnancy outcome.Conclusion:  Investigating the function of a gene during implantation in the mouse appears feasible using the jetPEI‐based method ofgene delivery. Targeted analyses of specific genes are in progress.
ABSTRACTS: 7 Study of uterine spiral arteries during implantation window in women with normal fertility or implantation failure
Authors: Mona Rahmati , Carine Munaut , Valerie Seravin , Sylvie Dubanchet , Corinne Morvan , Gerard Chaouat , Nathalie Lédée 
Published: June 2008

Problem:  Vascular remodelling and angiogenesis are essential for effective human implantation. Clinical tests measuring plasmatic angiogenic factors tend to predict complications of impaired implantation; however, the role of these factors in the endometrium at the time of implantation is not established and has been investigated.Material and Methods:  On patients with normal fertility or implantation failures, we performed pelvic 3‐D ultrasound and endometrial biopsy during mid‐luteal phase. Endometrial volume, resistance of uterine arteries and sub endometrial blood flow were quantified by ultrasound. For each biopsy, one sample was used for histological dating, CD56+ count, morphological study of vessels and immunohistochemistry (VEGF, VEGF‐R1, VEGF‐R2). Another sample underwent RNA extraction and RT‐PCR (angiogenic factors).Results:  We noticed that our data (morphology, immunohistochemistry, profile of pro‐angiogenic expression) tend to be different between our two groups.Conclusion:  These results may help establishing new criteria to define optimum uterine receptivity.
ABSTRACTS: 8 Identifying the receptor‐binding part of PIBF
Authors: Halasz Melinda , Polgar Beata , Kozma Noemi , Berta Gergely , Toth Gabor , Szekeres‐Bartho Julia 
Published: June 2008

Problem:  The PIBF1‐gene contains 18 exons and encodes a protein of 757 aminoacid residues with an 89‐kDa molecular mass. We aimed to identify the receptor‐binding region of the molecule, by its ability to activate signal transduction via JAK1/STAT6‐pathway in peripheral lymphocytes.Materials and Methods:  Recombinant PIBF constructs and small synthetic PIBF analogues representing different regions of the molecule were designed. EMSA was performed to detect nuclear translocation of STAT6‐dimers while intracellular STAT6‐phosphorylation was tested by Western blotting. PIBF‐receptor binding ability of the selected analogue was confirmed by confocal microscopy.Results:  Nuclear translocation of STAT6‐dimers was induced by PN1(exons 2‐3‐4), PN3(exons 8‐9), PN5(exons 3‐14‐15‐16) constructs. Peptide‐5 (in PN1) and 8 (in PN3‐4) were capable to activate STAT6‐phosphorylation. BodipyFL‐ labelled peptide‐8 bound to the PIBF‐receptor and was internalized in a time‐dependent manner.Conclusions:  These data suggest that the receptor‐binding region of PIBF might be a conformational structure that includes remote sequences.
ABSTRACTS: 9 Phenotype characterization of the anti‐inflammatory behavior of decidual endothelial cells
Authors: Agostinis C , Masat E , Bossi F , Lombardelli L , Tripodo C , Radillo O , De Seta F , Piccinni MP , Bulla R , Tedesco F 
Published: June 2008

Problem:  During pregnancy, an adequate inflammatory control is essential for normal intrauterine foetus development. Vascular endothelium is known to play a critical role in regulation of inflammatory processes. We hypothesize a different ability of decidual endothelial cells (DEC) compared to endothelium from other districts to respond to inflammatory stimuli.Material and Methods:  DEC were isolated and activated with inflammatory stimuli and their response in term of adhesion molecules expression, vascular leakage and cytokines production was evaluated. These data were compared to the data obtained with HUVEC and microvascular endothelial cells isolated from human skin (ADMEC) and from human endometrium (UtMEC).Results:  Our observations indicate that DEC: 1. express low levels of adhesion molecules after inflammatory stimulation 2. are unable to respond to classic permeabilizing stimuli 3. present a different cytokine profile.Conclusions:  Our results indicate that decidual endothelium has a different resting phenotype and shows an anti‐inflammatory behavior compared to other microvascular endothelium.
Attention profiles in autistic children with and without comorbid hyperactivity and attention problems
Authors: Sinzig Judith , Bruning Nicole , Morsch Dagmar , Lehmkuhl Gerd 
Published: June 2008

Objective:Psychopathological, neuropsychological and genetic findings indicate an association between ASD Spectrum Disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD). The goal of this study was to compare the neuropsychological profiles of attention functions in children with ADHD and with ASD and without comorbid ADHD. The hypothesis was that either ADHD and autistic children with comorbid ADHD symptoms were more impaired in inhibition and sustained attention performance and that all individuals with ASD show more deficits in divided attention.Method:Children aged 6 to 18 years old with ADHD (n = 30) or ASD with (n = 21) and without comorbid ADHD (n = 20) and 30 healthy children were included consecutively. Psychopathology was evaluated using the KIDDIE-SADS and symptom checklists for ADHD and ASD according to DSM-IV. Assessed neuropsychological functioning included inhibition, sustained as well as divided attention and alertness tasks.Results:Age and IQ-corrected z-scores were used. Statistically significant group effects were found for the variables sustained attention median (F = 3.2, = .02), hits (F = 3.3, p = .02) and false alarms (F = 3.9, p = .01), divided attention hits (F = 3.3, p = .02), errors (F = 3.1, p = .03) and false alarms (F = 3.3, p = .03) and alertness false alarms (F = 2.9, p = .04). Pearson Correlations revealed associations between ADHD symptoms and sustained attention in the ADHD group and between ADHD symptoms and inhibition in the ASD+ group.Conclusion:Our hypothesis was partly confirmed as ADHD children showed more deficits in sustained attention and ASD children in divided attention tasks. However there was no evidence that children with ASD and comorbid ADHD symptoms have a specific profile in comparison to pure ASD children.
Long-Term Safety and Efficacy of Tadalafil 5 mg Dosed Once Daily in Men with Erectile Dysfunction
Authors: Porst Hartmut , Rajfer Jacob , Casabé Adolfo , Feldman Robert , Ralph David , Vieiralves Luiz F. , Esler Anne , Wolka Anne M. , Klise Suzanne R. 
Published: June 2008

ABSTRACT

Introduction
With once-daily administration of tadalafil, dosing and sexual activity would no longer need to be temporally linked for patients with erectile dysfunction (ED).

Aim
To evaluate long-term safety and efficacy of tadalafil 5 mg dosed once daily for the treatment of ED.

Methods
Patients ≥18 years of age with ED of any functional severity or etiology received tadalafil 5 mg once daily for 1 (N = 234) or 2 (N = 238) years during the open-label extensions of two previously reported studies. Patients who completed the 1-year open-label extension concluded with a 4-week ED treatment-free period. Baseline was defined as prior to receiving any study drug.

Main Outcome Measures
Safety measures included adverse events, electrocardiograms, and clinical laboratory measures. Efficacy measures included the International Index of Erectile Function (IIEF)-Erectile Function (-EF), -Intercourse Satisfaction (-IS), and -Overall Satisfaction (-OS) domain scores, and the Global Assessment Questions (GAQ1: improved erections; GAQ2: improved ability to engage in sexual activity).

Results
Overall, 208/234 (88.9%) and 139/238 (58.4%) patients completed the 1- and 2-year open-label extensions, respectively. No study drug-related serious adverse events were observed. Treatment-emergent adverse events observed in ≥5% of the patients during the first year of either open-label extension were dyspepsia, headache, back pain, and influenza. No clinically meaningful abnormalities associated with tadalafil were observed for electrocardiograms or clinical laboratory measures. Mean IIEF domain scores improved from baseline to the conclusions of the 1- and 2-year open-label extensions, respectively: -EF, +10.4 and +10.8; -IS, +4.0 and +3.7; and -OS, +3.0 and +3.2. At the conclusion of the 2-year open-label extension, 95.7% and 92.1% of the patients reported positive responses to GAQ1 and GAQ2, respectively.

Conclusions
In these long-term, open-label, once-daily dosing studies, tadalafil 5 mg was well tolerated and effective, making it a viable alternative to the current on-demand dosing of tadalafil for men with ED.
Effects of Abiotic Stressors on Synthesis of the Mycosporine‐like Amino Acid Shinorine in the Cyanobacterium Anabaena variabilis PCC 7937
Authors: Singh Shailendra P. , Klisch Manfred , Sinha Rajeshwar P. , Häder Donat‐P. 
Published: June 2008

AbstractIn the present investigation we show that the cyanobacterium Anabaena variabilis PCC 7937 produces a single mycosporine‐like amino acid (MAA), shinorine (retention time = 2.3 min and absorption maximum at 334 nm) when isolated and purified by HPLC. Although there was significant induction of MAA synthesis from its initial value under 395 or 320 nm cutoff filters, MAA induction was significantly more pronounced in samples covered with 295 nm cutoff filters after 72 h of exposure. Heat as a stress factor had no effect on MAA induction with or without UV radiation. In contrast, salt and ammonium treatment had synergistic effects with UV stress. MAA synthesis was also induced by salt and ammonium in a concentration‐dependent manner without UV stress in samples covered with 395 nm cutoff filters. The results indicate that MAAs may have other functions in addition to photoprotection in this organism.
A Role for Internal Water Molecules in Proton Affinity Changes in the Schiff Base and Asp85 for One‐way Proton Transfer in Bacteriorhodopsin†
Authors: Morgan Joel E. , Gennis Robert B. , Maeda Akio 
Published: June 2008

AbstractLight‐induced proton pumping in bacteriorhodospin is carried out through five proton transfer steps. We propose that the proton transfer to Asp85 from the Schiff base in the L‐to‐M transition is accompanied by the relocation of a water cluster on the cytoplasmic side of the Schiff base from a site close to the Schiff base in L to the Phe219‐Thr46 region in M. The water cluster present in L, formed at 170 K, is more rigid than that at room temperature. This may be responsible for blocking the conversion of L to M at 170 K. In the photocycle at room temperature, this water cluster returns to the site close to the Schiff base in N, with a rigid structure similar to that of L at 170 K. The increase in the proton affinity of Asp85, which is a prerequisite for the one‐way proton transfer in the M‐to‐N transition, is suggested to be facilitated by a structural change which disrupts interactions between Asp212 and the Schiff base, and between Asp212 and Arg82. We propose that this liberation of Asp212 is accompanied by a rearrangement of the structure of water molecules between Asp85 and Asp212, stabilizing the protonated Asp85 in M.