Uranous ion oxidation and carbon dioxide fixation byThiobacillus ferrooxidans

The stoichiometric oxidation of uranous-to uranyl-uranium byThiobacllus ferrooxidans is demonstrated. Fixation of¹⁴CO₂ and the effect of inhibitors demonstrate that energy is conserved during the oxidation and used for energy-dependent reverse electron flow and carbon dioxide fixation.Abbreviations HOQNO 2-heptyl-4-hydroxyquinoline-N-oxide - 8-HQ 8-hydroxyquinoline - TTFA thenoyltrifluoroacetone     

An endo-1,4-β-glucanase expressed at high levels in rapidly expanding tissues

Plant developmental processes involving modifications to cell wall structure, such as cell expansion, organ abscission and fruit ripening, are accompanied by increased enzyme activity and mRNA abundance of endo-1,4--glucanases (EGases). An EGase cDNA clone, Ce14, isolated from tomato (Lycopersicon esculentum) has been shown to be identical to a tomato pistil-predominant EGase cDNA, TPP18. In addition to its previously reported expression during certain stages of early pistil development, Ce14 mRNA was also detected at high levels in the growing zones of etiolated hypocotyls (about 2.5-fold less than in pistils) and in young expanding leaves (about 3.5-fold less than in pistils). The abundance of Ce14 mRNA declined precipitously in older tissues as cells became fully expanded, and was barely detectable in mature vegetative tissues. Ce14 mRNA abundance was also low in abscission zones, and did not increase as abscission progressed. In fruit, Ce14 mRNA was present at low levels during fruit expansion, but was essentially absent during subsequent fruit development and ripening. Treatment of etiolated hypocotyls with ethylene or high concentrations of auxin sufficient to induce rapid lateral cell expansion and hypocotyl swelling also brought about an approximate doubling of Ce14 mRNA abundance, suggesting that Ce14 mRNA accumulation may be promoted directly or indirectly by ethylene. Thus, accumulation of Ce14 mRNA was found to be correlated with rapid cell expansion in pistils, hypocotyls and leaves.     

Alterations in growth and body composition during puberty. I.Comparing multicompartment body composition models

Roemmich, James N., Pamela A. Clark, Arthur Weltman, andAlan D. Rogol. Alterations in growth and body composition duringpuberty. I. Comparing multicompartment body composition models.J. Appl. Physiol. 83(3): 927-935, 1997.A four-compartment (4C) model of body composition was used as acriterion to determine the accuracy of three-compartment (3C) andtwo-compartment (2C) models to estimate percent body fat (%BF) inprepubertal and pubertal boys (genital I & II,n = 17; genital III & IV,n = 7) and girls (breast I & II, n = 8; breast III & IV,n = 15). The 3C water-density (3C-H₂O) and 3C mineral-densitymodels, dual-energy X-ray absorptiometry, the Lohman age-adjustedequations, the Slaughter et al. skinfold equations, and the Houtkooperet al. and Boileau bioelectrical impedance equations wereevaluated. Agreement with the 4C model increased with thenumber of compartments (i.e., body water, bone mineral) measured.Except for the 3C-H₂O model, thelimits of agreement were large and did not perform well forindividuals. The mean %BF by dual-energy X-ray absorptiometry (23.6%)was greater than that of the criterion 4C method (21.7%).For the field methods, the Slaughter et al. skinfold equationsperformed better than did the Houtkooper et al. and Boileaubioimpedance equations. The hydration of the fat-free mass decreased(genital I & II = 75.7%, genital III & IV = 74.8%, breast I & II = 75.5%, breast III & IV = 74.4%) and the mineral content increased(genital I & II = 4.9%, genital III & IV = 5.0%, breast I & II = 5.1%, breast III & IV = 5.7%) with maturation. The densityof the fat-free mass also increased (genital I & II = 1.084 g/ml,genital III & IV = 1.087 g/ml, breast I & II = 1.086 g/ml, breast III & IV = 1.091 g/ml) with maturation. All of the models reduced the %BF overprediction of the Siri 2C model, but only the 4C and3C-H₂O models should be used ascriterion methods for body composition validation in children andadolescents.

     

Evaluation of cDNA Libraries from Different Developmental Stages of Schistosoma mansoni for Production of Expressed Sequence Tags (ESTs)

A comparative study of the gene expression profile in differentdevelopmental stages of Schistosoma mansoni has been initiatedbased on the expressed sequence tag(EST) approach. A total of1401 ESTs were generated from seven different cDNA librariesconstructed from four distinct stages of the parasite life cycle.The libraries were first evaluated for their quality for a large-scalecDNA sequencing program. Most of them were shown to have lessthan 20% useless clones and more than 50% new genes. The redundancyof each library was also analyzed, showing that one adult wormcDNA library was composed of a small number of highly frequentgenes. When comparing ESTs from distinct libraries, we coulddetect that most genes were present only in a single library,but others were expressed in more than one developmental stageand may represent housekeeping genes in the parasite. When consideringonly once the genes present in more than one library, a totalof 466 unique genes were obtained, corresponding to 427 newS. mansoni genes. From the total of unique genes, 20.2% wereidentified based on homology with genes from other organisms,8.3% matched S. mansoni characterized genes and 71.5% representunknown genes.     

Functional studies in living animals using multiphoton microscopy

In vivo microscopy is a powerful method for studying fundamental issues of physiology and pathophysiology. The recent development of multiphoton fluorescence microscopy has extended the reach of in vivo microscopy, supporting high-resolution imaging deep into the tissues and organs of living animals. As compared with other in vivo imaging techniques, multiphoton microscopy is uniquely capable of providing a window into cellular and subcellular processes in the context of the intact, functioning animal. In addition, the ability to collect multiple colors of fluorescence from the same sample makes in vivo microscopy uniquely capable of characterizing up to three parameters from the same volume, supporting powerful correlative analyses. Since its invention in 1990, multiphoton microscopy has been increasingly applied to numerous areas of medical investigation, providing invaluable insights into cell physiology and pathology. However, researchers have only begun to realize the true potential of this powerful technology as it has proliferated beyond the laboratories of a relatively few pioneers. In this article we present an overview of the advantages and limitations of multiphoton microscopy as applied to in vivo imaging. We also review specific examples of the application of in vivo multiphoton microscopy to studies of physiology and pathology in a variety of organs including the brain, skin, skeletal muscle, tumors, immune cells, and visceral organs.     

Cytochrome P450-type hydroxylation and epoxidation in a tyrosine-liganded hemoprotein, catalase-related allene oxide synthase

The ability of hemoproteins to catalyze epoxidation or hydroxylation reactions is usually associated with a cysteine as the proximal ligand to the heme, as in cytochrome P450 or nitric oxide synthase. Catalase-related allene oxide synthase (cAOS) from the coral Plexaura homomalla, like catalase itself, has tyrosine as the proximal heme ligand. Its natural reaction is to convert 8R-hydroperoxy-eicosatetraenoic acid (8R-HPETE) to an allene epoxide, a reaction activated by the ferric heme, forming product via the Fe(IV)-OH intermediate, Compound II. Here we oxidized cAOS to Compound I (Fe(V)=O) using the oxygen donor iodosylbenzene and investigated the catalytic competence of the enzyme. 8R-hydroxyeicosatetraenoic acid (8R-HETE), the hydroxy analog of the natural substrate, normally unreactive with cAOS, was thereby epoxidized stereospecifically on the 9,10 double bond to form 8R-hydroxy-9R,10R-trans-epoxy-eicosa-5Z,11Z,14Z-trienoic acid as the predominant product; the turnover was 1/s using 100 μm iodosylbenzene. The enantiomer, 8S-HETE, was epoxidized stereospecifically, although with less regiospecificity, and was hydroxylated on the 13- and 16-carbons. Arachidonic acid was converted to two major products, 8R-HETE and 8R,9S-eicosatrienoic acid (8R,9S-EET), plus other chiral monoepoxides and bis-allylic 10S-HETE. Linoleic acid was epoxidized, whereas stearic acid was not metabolized. We conclude that when cAOS is charged with an oxygen donor, it can act as a stereospecific monooxygenase. Our results indicate that in the tyrosine-liganded cAOS, a catalase-related hemoprotein in which a polyunsaturated fatty acid can enter the active site, the enzyme has the potential to mimic the activities of typical P450 epoxygenases and some capabilities of P450 hydroxylases.     

When a virus is not a parasite: the beneficial effects of prophages on bacterial fitness

Most organisms on the planet have viruses that infect them. Viral infection may lead to cell death, or to a symbiotic relationship where the genomes of both virus and host replicate together. In the symbiotic state, both virus and cell potentially experience increased fitness as a result of the other. The viruses that infect bacteria, called bacteriophages (or phages), well exemplify the symbiotic relationships that can develop between viruses and their host. In this review, we will discuss the many ways that prophages, which are phage genomes integrated into the genomes of their hosts, influence bacterial behavior and virulence.     

Genome scans for transmission ratio distortion regions in mice

Transmission ratio distortion (TRD) is the departure from the expected genotypic frequencies under Mendelian inheritance. This departure can be due to multiple physiological mechanisms during gametogenesis, fertilization, fetal and embryonic development, and early neonatal life. Although a few TRD loci have been reported in mouse, inheritance patterns have never been evaluated for TRD. In this article, we developed a Bayesian binomial model accounting for additive and dominant deviation TRD mechanisms. Moreover, this model was used to perform genome-wide scans for TRD quantitative trait loci (QTL) on six F2 mouse crosses involving between 296 and 541 mice and between 72 and 1854 genetic markers. Statistical significance of each model was checked at each genetic marker with Bayes factors. Genome scans revealed overdominance TRD QTL located in mouse chromosomes 1, 2, 12, 13, and 14 and additive TRD QTL in mouse chromosomes 2, 3, and 15, although these results did not replicate across mouse crosses. This research contributes new statistical tools for the analysis of specific genetic patterns involved in TRD in F2 populations, our results suggesting a relevant incidence of TRD phenomena in mouse with important implications for both statistical analyses and biological research.     

Lymphatic function is regulated by a coordinated expression of lymphangiogenic and anti-lymphangiogenic cytokines

Lymphangiogenic cytokines such as vascular endothelial growth factor-C (VEGF-C) are critically required for lymphatic regeneration; however, in some circumstances, lymphatic function is impaired despite normal or elevated levels of these cytokines. The recent identification of anti-lymphangiogenic molecules such as interferon-γ (IFN-γ), transforming growth factor-β1, and endostatin has led us to hypothesize that impaired lymphatic function may represent a dysregulated balance in the expression of pro/anti-lymphangiogenic stimuli. We observed that nude mice have significantly improved lymphatic function compared with wild-type mice in a tail model of lymphedema. We show that gradients of lymphatic fluid stasis regulate the expression of lymphangiogenic cytokines (VEGF-A, VEGF-C, and hepatocyte growth factor) and that paradoxically the expression of these molecules is increased in wild-type mice. More importantly, we show that as a consequence of T-cell-mediated inflammation, these same gradients also regulate expression patterns of anti-lymphangiogenic molecules corresponding temporally and spatially with impaired lymphatic function in wild-type mice. We show that neutralization of IFN-γ significantly increases inflammatory lymph node lymphangiogenesis independently of changes in VEGF-A or VEGF-C expression, suggesting that alterations in the balance of pro- and anti-lymphangiogenic cytokine expression can regulate lymphatic vessel formation. In conclusion, we show that gradients of lymphatic fluid stasis regulate not only the expression of pro-lymphangiogenic cytokines but also potent suppressors of lymphangiogenesis as a consequence of T-cell inflammation and that modulation of the balance between these stimuli can regulate lymphatic function.