T M Mayhew

..This chapter reviews these new methods and illustrates their application with a consistent dataset...

..The aim of the current studies was to investigate whether these effects of estradiol are direct effects in the testis, or indirect actions via paradoxical stimulation of FSH secretion from the pituitary gland...

..The decline in total conductances is indicative of perturbations operating at the levels of villous trophoblast and fetal vasculature and these may contribute to fetal hypoxic stress...

..The latter is due to production of fewer and/or shorter capillary segments (rather than a decrease in capillary calibre), does not affect villous capillarization and is not accompanied by luminal remodelling...

..Its application is illustrated using datasets from immunolabelling studies designed to localise the KDEL receptor, phosphatidyl-inositol 4,5-bisphosphate, GLUT4 and rab4 at the electron microscopic level...

..The various methods are illustrated with worked examples based on empirical and synthetic data and will be of practical benefit to those applying single or multiple immunogold labelling in their research...

..Examples include diffusive transport, villous growth, fetoplacental angiogenesis, trophoblast turnover, arterial vascular remodelling and high-resolution immuno-localization studies...

..The method is illustrated using datasets from immunolabelling studies on the localization of GLUT1 glucose transporters in cultured human trophoblast cells exposed to different treatments...

..A more integrated approach involving also parallel analysis of the effects of erythropoietin and other potential vasoactive factors on the behaviour and morphology of fetal vessels would be beneficial...

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..Allometric analysis of results from recent studies on the murine placenta suggest further that diffusive conductances may also be matched to fetal mass during gestation and to fetal mass at term across species...

..It is hoped that this review will inform future study designs, encourage greater transparency and facilitate sampling improvements...

..We conclude that finger lengths are cycle-dependent and that account should be taken of this, and of oral contraceptive usage, in future studies on female digit lengths and their ratios...

..g. total volumes, surfaces and numbers of structures per cell. Here, the utility, limitations and recent applications of these methods are reviewed...

..LDs of the chosen compartments are used to calculate RLI and chi-squared values and these serve to identify those compartments in which there is preferential labelling. The methods are illustrated by synthetic and real data...

..Finally, we discuss labelling efficiency (the number of gold particles per target molecule) and describe how it can be estimated for volume- or surface-occupiers by combining stereological data with biochemical determinations...

..Finally, quantitative immunoelectron microscopy now permits more rigorous analysis of the spatial distributions of interesting molecules between subcellular compartments or shifts in distributions following experimental manipulation...

..The methods are illustrated using data taken from studies aimed at localising protein antigens (caveolin-1 and GLUT1) in specimens of term human placenta...

..These simple and efficient approaches should be of more immediate utility to those interested in codistribution and colocalization in multiple immunogold labelling investigations...

..This activity seems to reside in thin regions of syncytium...

..This approach provides a simple and efficient way of comparing LDs in different compartments. Its utility is illustrated using data from VPARP and LAMP-1 labelling experiments...

..These changes are likely to reduce placental oxygen diffusive conductances and contribute to fetal hypoxic stress...

..Therefore, it is likely that fetal hypoxia associated with maternal diabetes mellitus is due to metabolic disturbances rather than abnormalities in the quantities or arrangements of maternal vascular spaces...

..The nascent detachment site is immediately covered by FTF prior to repair by re-epithelialization...

..These studies re-emphasize the relative invariance of biological organization and the value and limitations of covariance analysis for quantifying different levels of organization in different tissues and experimental groups...

..We propose further that clustered nuclei (pre-apoptotic and apoptotic) in syncytial knots probably represent the extrusion component of normal continuous epithelial turnover...

..These findings provide quantitative confirmation of the polarised arrangement of zymogen granules within acinar cells and further support for the relative invariance of biological organisation between subjects...

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..To test the hypothesis that thinning of the placental intervascular layers, and greater variability in thickness, are positive adaptations to facilitate passive diffusion and may help to resolve different categories of fetal hypoxia...

..7 million nuclei per hour) and that for endothelial cell nuclei was 5.1 x 10(7) per week (0.3 million nuclei per hour). Predictions are made about the postnatal ages at which adult ratios of different nuclear types might be attained...

..It is concluded that normal values are preserved by good glycaemic control regardless of diabetic grouping...

..Elaboration of finer terminal branches on villous trees leads to a decline in the star volumes of villi and intervillous pores. Some of the functional implications of these findings are discussed...

..Adaptive re-settings of the epithelial steady state may also occur in abnormal pregnancies...

..Here, placentas from control subjects and patients with pre-gestational Type I (insulin-dependent) diabetes mellitus were used to test for differences in measures of angiogenesis and vascular remodelling...

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..The importance of controlling for various potential confounders is also emphasized...

..Sometimes (e.g. when relying on biopsy samples), the size of the reference cannot be determined. In such cases, ratio data must be interpreted with due caution...

..The findings are partly consistent with results from in vitro studies which indicate that hypoxia stimulates proliferation of cytotrophoblast but impairs fusion into syncytium...

..Contingency table analysis allows labelling distributions in different groups of cells to be compared. Protocols are described and illustrated using worked specimen examples and real data...

..Here, the current state of the art is reviewed and approaches are illustrated with virtual datasets...

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..The findings suggest that the morphometric differences in fetoplacental vascularity are likely to be due to the effects of asthma and use of inhaled GCs rather than the effects of maternal or fetal hypoxic stress...

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..5. This study provides conclusive evidence that administration of TNP-470 interferes with placental vascular proliferation and vessel caliber and results in a reproducible model of IUGR...

..Alterations in these vessels may contribute to increased uterine vascular resistance in preeclampsia...

..If more precise estimates of labeling proportions over individual compartments are required (e.g., to achieve coefficients of error of 10-20%), then 100-200 particles need to be counted over each compartment of interest...

..We first describe the preconditions and the way to implement these methods, then provide three worked examples, and finally discuss the advantages, pitfalls, and limitations of this method...