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Hormone measures in finger-prick blood spot samples: new field methods for reproductive endocrinology.

Comparative endocrine studies have notably advanced understanding of ecological factors that contribute to variation in human reproductive function. Such research has relied on methodological advances that permit hormone determinations in samples that are easily and safely collected, stored, and transported, most recently on measurement of steroids in saliva. This report seeks to further expand the scope of endocrine research by demonstrating the value of blood spot samples collected by finger prick. As a sampling strategy, finger-prick blood spot collection offers the advantages of short collection time, low invasiveness, repeatability, absence of postcollection processing, low biohazard risk, and ease of sample storage and transport. We document good sample stability and present sensitive assay methods for a range of steroids and proteins (FSH, LH, PRL, T, E2, DHEAS, androstenedione, cortisol, SHGB) in blood spots that require sample volumes of 3-12 microliters and display good reliability, specificity, precision, accuracy, and convertibility of results to plasma/serum equivalent concentrations. Laboratory evaluation was augmented by a feasibility study at a remote site in Papua New Guinea that confirmed validity and stability of blood spot collections under field conditions. Research applications of blood spot sampling are illustrated with a series of studies, including cross-sectional surveys for developmental and life span endocrinology, a longitudinal, population-based developmental epidemiologic study of puberty, and serial sampling in a dynamic study of neuroendocrine response to suckling. We conclude that the sampling features and wide range of measurable biomolecules of blood spots do constitute a methodological advance for endocrine research.

Endocrinology↗

Characterization of melanocyte-specific inducible Cre recombinase transgenic mice.

Conditional Cre-mediated recombination has emerged as a robust method of introducing somatic genetic alterations in an organ-specific manner in the mouse. Here, we generated and characterized mice harboring a 4-hydroxytamoxifen (OHT)-inducible Cre recombinase-estrogen receptor fusion transgene under the control of the melanocyte-specific tyrosinase promoter, designated Tyr::CreER(T2). Cre-mediated recombination was induced in melanocytes in a spatially and temporally controlled manner upon administration of OHT and was documented in embryonic melanoblasts, follicular bulb melanocytes, dermal dendritic melanocytes, epidermal melanocytes of tail skin, and in putative melanocyte stem cells located within the follicular bulge. Functional evidence suggestive of recombination in follicular melanocyte stem cells included the presence of Cre-mediated recombination in follicular bulb melanocytes 1 year after topical OHT administration, by which time several hair cycles have elapsed and the melanocytes residing in this location have undergone multiple rounds of apoptosis and replenishment. These Tyr:: CreER(T2) transgenic mice represent a useful resource for the evaluation of melanocyte developmental genetics, the characterization of melanocyte stem cell function and dynamics, and the construction of refined mouse models of malignant melanoma.

Animals↗

Force adaptation transfers to untrained workspace regions in children: evidence for developing inverse dynamic motor models.

When humans perform goal-directed arm movements under the influence of an external damping force, they learn to adapt to these external dynamics. After removal of the external force field, they reveal kinematic aftereffects that are indicative of a neural controller that still compensates the no longer existing force. Such behavior suggests that the adult human nervous system uses a neural representation of inverse arm dynamics to control upper-extremity motion. Central to the notion of an inverse dynamic model (IDM) is that learning generalizes. Consequently, aftereffects should be observable even in untrained workspace regions. Adults have shown such behavior, but the ontogenetic development of this process remains unclear. This study examines the adaptive behavior of children and investigates whether learning a force field in one hemifield of the right arm workspace has an effect on force adaptation in the other hemifield. Thirty children (aged 6-10 years) and ten adults performed 30 degrees elbow flexion movements under two conditions of external damping (negative and null). We found that learning to compensate an external damping force transferred to the opposite hemifield, which indicates that a model of the limb dynamics rather than an association of visited space and experienced force was acquired. Aftereffects were more pronounced in the younger children and readaptation to a null-force condition was prolonged. This finding is consistent with the view that IDMs in children are imprecise neural representations of the actual arm dynamics. It indicates that the acquisition of IDMs is a developmental achievement and that the human motor system is inherently flexible enough to adapt to any novel force within the limits of the organism's biomechanics.

Adaptation, Physiological↗

An event-related potentials study of biological motion perception in human infants.

To clarify the dynamical processing aspect of biological motion (BM) perception from a developmental point of view, we measured event-related potentials (ERPs) in 8-month-old infants during the perception of BM or a scrambled motion (SM; randomization of BM's spatial structure). We found that activation of the right hemisphere in 8-month-old infants was similar to that of adults, suggesting that the neural substrates for processing BM perception begin to mature at around 8 months of age.

Analysis of Variance↗

Nuclear magnetic resonance imaging in central nervous system disease.

From the preliminary work of many investigators, it appears that proton nuclear magnetic resonance (NMR) imaging will have wide application in the diagnostic assessment (and potential management) of patients with vascular, neoplastic, and demyelinating diseases of the central nervous system (CNS). Findings in isolated cases and small series suggest that NMR imaging may play a role in the evaluation of patients with other CNS conditions including hydrocephalus, malformations, infections, developmental and metabolic disorders, and degenerative processes. Because of the dynamic nature of disease processes involving the CNS, the precise meaning of NMR image parameters (rho, T1, and T2) remains unclear. A comprehensive study correlating NMR images in neurologic disease with precise neuropathologic examination is required. In the future, with accurate quantitative measurements of these NMR parameters, in vivo imaging may provide insight into the dynamic nature of neurologic disease.

Central Nervous System Diseases↗

Peripheral and central factors limiting the development of contrast sensitivity in macaque monkeys.

The aim of this study was to evaluate the contribution of peripheral and central factors to the development of visual sensitivity. We used the approach of (Pelli, 1981, 1990) to evaluate the hypothesis that intrinsic noise is high in infants compared with adults, and therefore sets an important limit on contrast sensitivity in infants. We measured contrast thresholds in the presence of various levels of dynamic spatiotemporal broadband noise in infant monkeys, and evaluated the developmental changes in contrast threshold and intrinsic noise. Our data show that intrinsic noise is high in infants and falls with contrast threshold during development. However, contrast thresholds in high-contrast noise also fall during development, although by a smaller amount. Therefore, while changes in intrinsic noise set an important limit on the development of contrast sensitivity across spatial frequencies, changes in non-additive sources of noise also contribute, particularly at high spatial frequencies. We interpret these results in terms of Pelli's hypothesis about the sources of additive and non-additive noise affecting visual detection. In these terms, additive noise reflects peripheral factors and non-additive noise reflects central ones. Our results suggest that changes in peripheral sources of noise represent an important limit for the development of visual sensitivity.

Aging↗

Dynamic restricted expression of the chaperone Hsc70 in early chick development.

The non-inducible chaperone heat shock cognate 70 kDa (Hsc70) is regulated during development. We now characterize its dynamic expression pattern from gastrulation to early organogenesis. Throughout this developmental period, hsc70 transcripts were largely restricted to neuroectoderm- and mesoderm-derived structures. In stage 10 embryos, Hsc70 protein was expressed in the neural tube with increasing rostrocaudal and decreasing dorsoventral gradients, and in some somite cells. This highly regulated expression of Hsc70 is likely to reflect specific developmental functions, besides its well-characterized role in protein folding.

Animals↗

The ins and outs of endocytic transport.

A fusion of cutting-edge research in cell biology, developmental biology and immunology made the recent workshop on Membrane Dynamics in Endocytosis an outstanding success. Members of an increasingly diverse community converged upon the small town of Sant Feliu de Guixols on the coast of Spain, between September 17-22, 2005, to discuss common themes emerging from their studies on membrane transport. Organized by Margaret Robinson (Cambridge, UK) and Howard Riezman (Geneva, Switzerland), the meeting covered diverse topics that highlighted essential roles for endocytosis during cell growth, development and parasitic invasion.

Animals↗

Desynchronization of cells on the developmental path triggers the formation of spiral waves of cAMP during Dictyostelium aggregation.

Whereas it is relatively easy to account for the formation of concentric (target) waves of cAMP in the course of Dictyostelium discoideum aggregation after starvation, the origin of spiral waves remains obscure. We investigate a physiologically plausible mechanism for the spontaneous formation of spiral waves of cAMP in D. discoideum. The scenario relies on the developmental path associated with the continuous changes in the activity of enzymes such as adenylate cyclase and phosphodiesterase observed during the hours that follow starvation. These changes bring the cells successively from a nonexcitable state to an excitable state in which they relay suprathreshold cAMP pulses, and then to autonomous oscillations of cAMP, before the system returns to an excitable state. By analyzing a model for cAMP signaling based on receptor desensitization, we show that the desynchronization of cells on this developmental path triggers the formation of fully developed spirals of cAMP. Developmental paths that do not correspond to the sequence of dynamic transitions no relay-relay-oscillations-relay are less able or fail to give rise to the formation of spirals.

Animals↗

Dynamic visualization of nervous system in live Drosophila.

We have constructed transgenic Drosophila melanogaster lines that express green fluorescent protein (GFP) exclusively in the nervous system. Expression is controlled with transcriptional regulatory elements present in the 5' flanking DNA of the Drosophila Na(+), K(+)-ATPase beta-subunit gene Nervana2 (Nrv2). This regulatory DNA is fused to the yeast transcriptional activator GAL4, which binds specifically to a sequence motif termed the UAS (upstream activating sequence). Drosophila lines carrying Nrv2-GAL4 transgenes have been genetically recombined with UAS-GFP (S65T) transgenes (Nrv2-GAL4+UAS-GFP) inserted on the same chromosomes. We observe strong nervous system-specific fluorescence in embryos, larvae, pupae, and adults. The GFP fluorescence is sufficiently bright to allow dynamic imaging of the nervous system at all of these developmental stages directly through the cuticle of live Drosophila. These lines provide an unprecedented view of the nervous system in living animals and will be valuable tools for investigating a number of developmental, physiological, and genetic neurobiological problems.

Animals↗

Evolution of virulence of a plant RNA virus in developmental stage-structured host populations.

Natural host populations are age-structured, and developmental stages differ in susceptibility and within-host pathogen dynamics, potentially imposing distinct selective pressures on viruses. However, the evolutionary consequences of host age structure remain poorly understood. We experimentally evolved turnip mosaic potyvirus for 5 passages in Arabidopsis thaliana populations spanning 7 demographic regimes, from juvenile- to mature-dominated cohorts. We quantified disease progression, symptom severity, and viral load, cross-inoculated evolved lineages across host stages to construct infection matrices, and performed whole-population sequencing at passages 1 and 5. Disease traits changed markedly with passage, demography, and their interaction. Disease progression evolved faster in older populations, whereas symptom severity was independent of median age, indicating demographic reweighting of virulence components. Viral load increased across passages and positively correlated with severity, linking within-host fitness to symptoms. Cross-inoculation assays revealed a modular infection network: juvenile-evolved lineages specialized on juvenile hosts, whereas lineages from intermediate and older populations were more generalist. Genomically, we detected both parallel and demography-specific adaptations, including recurrent changes in the viral protein VPg (involved in translation, replication, and host interactions) as well as synonymous variants showing consistent or opposing selection across host population stage structures. Overall, host age structure emerges as a major ecological driver of virulence evolution, shaping tradeoffs between disease progression and severity and determining specialization versus generalism. These results integrate phenotypic and genomic responses and suggest that manipulating crop age structure could steer virus evolution toward less damaging outcomes.

Virulence↗

Fetal lung development: compressibility as a measure of maturity.

Fetal lung maturation involves a number of separate developmental processes. An ultrasonic technique is reported for grading the dynamic behavior of the right middle lobe or lingula in the second or third trimesters on ultrasound (US) study. Fetal lungs are stiff initially. Lung compressibility during diastole is typical after 36 weeks' gestational age, when phospholipid profiles of the amniotic fluid also indicate lung-tissue maturity. Compressibility also occurs earlier in the third trimester and has indicated, in preliminary observations on a small subgroup of infants delivered prematurely, a low risk of subsequent neonatal respiratory distress syndrome. High-speed magnification US study of fetal lung compressibility can be used to evaluate fetal lung development, possibly independently of gestational age and of conventionally measured lecithin/sphingomyelin values in amniotic fluid.

Female↗

A milestone for normal development of the infantile brain detected by functional MRI.

OBJECTIVE: To investigate the relationship between cerebral cortical function and white matter myelination in the visual pathway in the evaluation of normal brain development. METHODS: The authors performed quantitative analysis of white matter myelination detected with conventional T1-weighted spin echo (SE) MRI and brain functional MRI (fMRI) using echoplanar imaging with photic stimulation in 27 neurologically normal infants (age range, 0 to 22 weeks). RESULTS: An age-dependent gradual increase in signal intensity was observed in optic radiation on the T1-weighted SE images, indicating progression of white matter myelination. A rapid age-dependent reverse in signal response was observed on fMRI. Infants older than 8 weeks showed a stimulus-induced signal decrease in the visual cortex, whereas infants younger than 7 weeks showed a signal increase. CONCLUSIONS: A rapid inversion of response revealed by fMRI with photic stimulation in infants suggests a change in oxygen consumption during neuronal activation, which is related to rapid synapse formation and accompanying increased metabolism. fMRI can detect dynamic metabolic changes during brain maturation, which is a different developmental process from white matter myelination. The metabolic changes detected by fMRI provide a milestone for the evaluation of normal brain development.

Brain↗

A probabilistic model of mosaicism based on the histological analysis of chimaeric rat liver.

The analysis of pattern development in mosaic and chimaeric animals has provided insight into a number of developmental problems. In order to aid the understanding of the dynamics of the development of mosaic tissues, a computer simulation of the generation of a mosaic tissue was created using simple probabilistic decisions. Results of quantitative analysis of the simulated mosaicism were compared with chimaeric liver. Chimaeric animals were produced by morula aggregation between histologically distinguishable strains of congenic rats. The livers of these animals revealed a pattern of patchy mosaicism unrelated to either acinar or lobular architecture of the organ. Independent quantifiable parameters were correlated and compared between the simulation and chimaeric liver tissue. This analysis showed that extensive cell migration is not required to develop finely variegated mosaic tissue and that the patterns of mosaicism observed could have resulted from tissue development in which as few as three reiterated decisions were required. First, the simulation established anlagen of two cell types of various specified proportions with randomly chosen placement. Second, in each generation of the simulation the order in which the cells divided was established randomly. Third, there was a random choice of the direction of placement of the daughter cell. The quantitative relationships between the proportion of cell types, the area of patches and the number of patches per unit area was consistent between the simulation and the chimaeric tissue.

Animals↗

Growth or differentiation? Adaptive regeneration in the brittlestar Amphiura filiformis.

Amphiura filiformis is a burrowing brittlestar, which extends arms in the water column when suspension feeding. In previous studies, unexpectedly high variability was observed in regeneration rate between individuals even when experiments were performed under identical conditions. The aims of this work were to understand this variability and interpret the observed variability in terms of adaptation to sublethal predation. Our experiments on the dynamics of arm regeneration in A. filiformis revealed that the developmental program during regeneration is well adapted to its burrowing life style. We demonstrate that there is a trade-off between regeneration in length and functional recovery for feeding (differentiation index). The amount of tissue lost (length lost), which represents the quantity of tissue needed to completely regenerate an intact arm with no previous history of regeneration, determines whether the arm will invest more energy in growth and/or in differentiation, which must be a reflection of the ability to differentially regulate developmental programs during regeneration. We show that combining regeneration rate with differentiation index provides an ideal tool for the definition of a standard temporal framework for both field and laboratory studies of regeneration.

Adaptation, Physiological↗

Three new Drosophila markers of intracellular membranes.

The need for cellular markers that permit a quick and accurate evaluation of a protein's subcellular localization has increased with the surge of new data generated by the Drosophila genome project. In this report, we present three ubiquitously expressed Drosophila transgenes that expressed a green fluorescent protein variant (enhanced yellow fluorescent protein) that has been targeted to different intracellular membrane targets: the Golgi apparatus, mitochondria, and endoplasmic reticulum. These markers serve as an internal standard for characterizing a protein's subcellular localization or as a means of tracking the dynamics of intracellular organelles during normal or abnormal cellular or developmental processes. We have also examined fixation artifacts using these constructs to illustrate the effects that fixation and permeabilization have on intracellular membrane organization.

Animals↗

Intramedullary rodding in type III osteogenesis imperfecta. Effects on neuromotor development in 10 children.

We studied retrospectively gross motor development and the impact of intramedullary rodding in 10 children with type III osteogenesis imperfecta (OI). There was a pronounced delay in motor development and the order in achieving gross motor milestones differed from the normal developmental sequence. Static milestones developed at an earlier stage than dynamic milestones. Intramedullary rodding of the lower extremities prior to the age of 3.5 years enhanced neuromotor development, especially regarding the milestones supported standing, rolling from prone to supine and crawling with abdomen on the floor. The different sequence in achieving gross motor milestones should have implications for future rehabilitation programs and for orthopedic surgery.

Body Height↗

Children with encopresis: A descriptive analysis.

During an 18-month period, 102 children with encopresis were seen in a general pediatric outpatient clinic. Through interviews and questionnaires, extensive historical information was obtained. Children were compared with respect to demographic characteristics, bowel-function histories, early training and management, and present manifestations of bowel dysfunction. A wide variety of historical backgrounds and manifestations was observed. It was found that many of the traditional generalizations about encopresis could not be substantiated. It is suggested that a very individualized approach to this problem be adopted and that children with encopresis be investigated carefully to uncover early developmental problems, critical life events, and the precise physiology and dynamics of present bowel dysfunction.

Adolescent↗