Age-dependent changes in porcine alveolar macrophage function during the postnatal period of alveolarization.

During early postnatal ontogeny in most mammals, the lung is structurally and functionally immature. In some species with relatively altricial lung morphology, there is evidence of a coupling between functional maturity of the pulmonary cellular immune system and alveolar maturation. Herein, we examine changes in alveolar macrophage (AM) number and function occurring during alveolarization in a more precocial species, the pig, to determine if heightened oxidative metabolism and phagocytic ability is similarly delayed until completion of lung morphogenesis. We assessed cell differential in lavage fluid and evaluated two main functional parameters of AM phagocytic response, the generation of reactive oxygen species (ROS), and particle internalization. AM functional maturation occurred mainly during the first postnatal week: the proportion of AMs, ROS generation, and phagocytosis all increased significantly. These results suggest maturational improvement of the impaired AM-based pulmonary immune system of the neonate piglet occurs during the postnatal period of rapid alveolarization.

Finite element 3D reconstruction of the pulmonary acinus imaged by synchrotron X-ray tomography.

The alveolated structure of the pulmonary acinus plays a vital role in gas exchange function. Three-dimensional (3D) analysis of the parenchymal region is fundamental to understanding this structure-function relationship, but only a limited number of attempts have been conducted in the past because of technical limitations. In this study, we developed a new image processing methodology based on finite element (FE) analysis for accurate 3D structural reconstruction of the gas exchange regions of the lung. Stereologically well characterized rat lung samples (Pediatr Res 53: 72-80, 2003) were imaged using high-resolution synchrotron radiation-based X-ray tomographic microscopy. A stack of 1,024 images (each slice: 1024 x 1024 pixels) with resolution of 1.4 mum(3) per voxel were generated. For the development of FE algorithm, regions of interest (ROI), containing approximately 7.5 million voxels, were further extracted as a working subunit. 3D FEs were created overlaying the voxel map using a grid-based hexahedral algorithm. A proper threshold value for appropriate segmentation was iteratively determined to match the calculated volume density of tissue to the stereologically determined value (Pediatr Res 53: 72-80, 2003). The resulting 3D FEs are ready to be used for 3D structural analysis as well as for subsequent FE computational analyses like fluid dynamics and skeletonization.

Effects of fluoride emissions from a modern primary aluminum smelter on a local population of white-tailed deer (Odocoileus virginianus).

The influence of fluoride emissions from a modern aluminum smelter on concentrations of skeletal fluoride and dental fluorosis in a resident population of white-tailed deer was studied. The smelter was located on Mount Holly Plantation in South Carolina, and concentrations of skeletal fluoride in the deer collected at Mount Holly increased approximately five-fold 3 yr after the operation began. Increases in skeletal fluoride of less than two-fold were observed in deer obtained from Medway Plantation which has its nearest boundary 1.6 km from the smelter site. No dental fluorosis was observed in deer collected at Medway Plantation, but mild dental fluorosis was observed in a significant number of deer collected at Mount Holly Plantation. The dental fluorosis that was observed was not associated with incisor wear or with fluoride-induced molar wear. Osteofluorosis of mandibles or metacarpals was not observed in any of the deer obtained from either plantation. The data obtained from this study indicated that the presence of a modern aluminum smelter caused a detectable increase in concentration of skeletal fluoride in the resident population of white-tailed deer, but that no adverse health effects were seen.

Kinetic output: A conceptual, dimensional and empirical analysis.

A dimensional analysis permitted behavioral frequency, duration and available time to be collapsed into a singular dimensionless expression called Kinetic output (K(o)). An empirical analysis across several behaviors in some human subjects showed that the derived index of behavioral output, K(o), was more likely to exhibit less variability than rate or duration measures of the behavior. As a reduction in variability is synonymous with finding an increased order, the results tentatively support the notion that behavior may exhibit conformity at a previously undetected level. One major implication of the K(o) formulation is that divergent frequency and duration performances are interpretable as reflecting alternate forms of the same equivalence.

Three-dimensional visualization of microvessel architecture of whole-mount tissue by confocal microscopy.

The three-dimensional architecture of the nascent microvascular network is a critical determinant of vascular perfusion in the setting of regenerative growth, vasculopathies and cancer. Current methods for microvessel visualization are limited by insufficient penetration and instability of endothelial immunolabels, inadequate vascular perfusion by the high-viscosity polymers used for vascular casting, and destruction of tissue stroma during the processing required for scanning electron microscopy. The aim of this study was to develop whole-mount tissue processing methods for 3D in situ visualization of the microvasculature that were also compatible with supplementary labeling for other structures of interest in the tissue microenvironment. Here, we present techniques that allow imaging of the microvasculature by confocal microscopy, to depths of up to 1500 mum below the specimen surface. Our approach includes labeling luminal surfaces of endothelial cells by i.v. injection of fluorescently conjugated lectin and filling the microvasculature with carbon or fluorescent nanoparticles/Mercox, followed by optical clearing of thick tissue sections to reduce light scatter and permit 3D visualization of microvessel morphology deep into the sample. Notably, tissue stroma is preserved, allowing simultaneous labeling of other structures by immunohistochemistry or nuclear dyes. Results are presented for various murine tissues including fat, muscle, heart and brain under conditions of normal health, as well as in the setting of a glioma model growing in the subcutaneous space or orthotopically in the brain parenchyma.

Oviduct structure and function and reproductive modes in amphibians.

The structure and function of the oviducts of members of the three Orders of the Class Amphibia (Anura, frogs and toads; Urodela, salamanders and newts; Gymnophiona, caecilians) are well described for only a few species. Further, the majority of such descriptions relate only to temperate species that breed in water, lay their eggs there, and have free-living larvae, the presumed ancestral condition of oviparity. Many species of amphibians have derived reproductive modes. Such modes include breeding terrestrially and arboreally, making foam nests, parental transport of eggs and/or tadpoles, direct development (copulating on land, laying the eggs in terrestrial sites, fully metamorphosed juveniles hatching, obviating the free-living larval stage). Other derived modes are ovoviviparity (developing embryos retained in the oviducts, born at a diversity stages of development, no maternal nutrition in addition to yolk) and viviparity (oviductal retention of developing young, maternal nutrition after yolk is resorbed, young born as fully metamorphosed juveniles). The amphibian oviduct is regionally differentiated to secrete varying numbers of layers of material around each egg, which function in fertilization, etc.; it is responsive to endocrine output and environmental mediation during the reproductive cycle; and it maintains developing embryos in some members of all three orders, some with oviductal epithelial secretion of nutrients. However, little is known of the structure-function relationships of the oviduct in species with derived reproductive modes. A comparison of oviduct morphology, function, endocrinology, ecology and phylogeny in amphibians with diverse reproductive modes suggests a number of highly productive avenues of investigation.