The role of the kidney in the biosynthesis of carnitine in the rat.

Intravenous administration of L-[methyl-3H]-labeled trimethyllysine to rats results in a very rapid accumulation of radioactivity by the kidneys, while the incorporation of the label into the liver occurs at approximately 1% of this rate when calculated per g of wet tissue. The kidneys convert a substantial portion of the trimethyllysine taken up to butyrobetaine and to beta-hydroxytrimethyllysine, a precursor of butyrobetaine, but fail to synthesize carnitine. Significant amounts of radioactivity are recovered in both carnitine and butyrobetaine of hepatic tissue after longer time periods, while the level of labeled trimethyllysine in this organ remains very low. Bilateral nephrectomy results in a marked decrease in the incorporation of label into the liver. These results indicate that in rats, the initial conversion of trimethyllysine to butyrobetaine occurs predominantly in kidney and that the liver capacity for this transformation is considerably smaller than its capacity to synthesize carnitine from butyrobetaine.     

The role of a beta-bulge in the folding of the beta-hairpin structure in ubiquitin

It is known that the peptide corresponding to the N-terminal beta-hairpin of ubiquitin, U(1-17), can populate the monomeric beta-hairpin conformation in aqueous solution. In this study, we show that the Gly-10 that forms the bulge of the beta-turn in this hairpin is very important to the stability of the hairpin. The deletion of this residue to desG10(1-16) unfolds the structure of the peptide in water. Even under denaturing conditions, this bulge appears to be important in maintaining the residual structure of ubiquitin, which involves tertiary interactions within the sequence 1 to 34 in the denatured state. We surmise that this residual structure functions as one of the nucleation centers in the folding process and is important in stabilizing the transition state. In accordance with this idea, deleting Gly-10 slows down the refolding and unfolding rate by about one half.     

Expression of Transferrin Binding Protein in the Capillaries of the Brain in the Developing Chick Embryo

Transferrin-binding protein (TfBP) has been shown to be a novel protein, structurally related to the chicken heat shock protein 108. The physiological function of this protein, however, has not yet been established. Antiserum to TfBP selectively stains transferrin- and iron-rich oligodendrocytes and choroidal epithelium in the adult and embryonic chick brain, suggesting a role for this protein in transferrin and iron storage in these cells. In this study, we further demonstrate TfBP-immunoreactivity (IR) in the blood vessels of the embryonic chick central nervous system. A strong TfBP-IR was present in blood vessels from E6, declined from E10 and was absent by E18. Thus, the expression of the TfBP in the blood vessels precedes its expression in the oligodendrocytes. At the subcellular level, TfBP-IR was confined to the cytoplasm of capillary pericytes while the Tf-receptor IR was associated with the capillary endothelium of the brain. The up-regulated expression of TfBP, together with the Tf-receptor of the brain capillaries, suggests that pericytes may be associated with the high iron uptake required for the metabolic demands of the developing brain. D. W. Kim and H. N. Lee contributed equally to this work.     

Cranial cartilages: Players in the evolution of the cranium during evolution of the chordates in general and of the vertebrates in particular

The present contribution is chiefly a review, augmented by some new results on amphioxus and lamprey anatomy, that draws on paleontological and developmental data to suggest a scenario for cranial cartilage evolution in the phylum chordata. Consideration is given to the cartilage-related tissues of invertebrate chordates (amphioxus and some fossil groups like vetulicolians) as well as in the two major divisions of the subphylum Vertebrata (namely, agnathans, and gnathostomes). In the invertebrate chordates, which can be considered plausible proxy ancestors of the vertebrates, only a viscerocranium is present, whereas a neurocranium is absent. For this situation, we examine how cartilage-related tissues of this head region prefigure the cellular cartilage types in the vertebrates. We then focus on the vertebrate neurocranium, where cyclostomes evidently lack neural-crest derived trabecular cartilage (although this point needs to be established more firmly). In the more complex gnathostome, several neural-crest derived cartilage types are present: namely, the trabecular cartilages of the prechordal region and the parachordal cartilage the chordal region. In sum, we present an evolutionary framework for cranial cartilage evolution in chordates and suggest aspects of the subject that should profit from additional study.     

The Role of the Root in the Induction of Xylem Differentiation in Peas

It is known that growing parts of the shoot induce the differentiationof vascular tissues below them and that this induction is dueto the production of auxin. The problem dealt with here is whythe formation of xylem proceeds in the growing roots. The redifferentiationof parenchyma to tracheary elements in grafts of pea plantswas used in this study. It is proved that this is not due tostimuli coming from the root tip but rather to the movementof a stimulus coming from the shoot into the root. The polarityof movement is maintained even in thin sections, but it canbe reversed by a strong shoot influence.     

Role of transposable elements in the propagation of minisatellites in the rice genome

A survey of minisatellites (MSs) in 5.3 Mb of randomly selected rice DNA sequences from public databases was carried out to clarify the role of transposable elements (TEs) in the dispersal of MSs in the rice genome. The estimated frequency of MSs in this sample was one per 23.4 kb, and this frequency is approximately equivalent to that of Class I microsatellites in the rice genome. Of the MSs in the 5.3-Mb sequence sample, 82% were found to be present in multiple copies in the rice genome, and all of these were a part of TE sequences. In this study at least 61 TE groups were identified as MS carriers. It was also shown that the GC-rich MS pOs6.2H, which was previously reported to be one of the interspersed MSs in the rice genome, is a component of an En / Spm -like element. These results indicate that the majority of MSs in the rice genome are maintained in TEs, and amplified and dispersed as components of the TEs. The G+C content of the multi-locus MS sequences reflected that of the TE sequences containing those MSs, but no obvious bias towards the high G+C content of DNA was observed. Single locus MSs also did not show any obvious bias towards the high G+C content of DNA in the rice genome. In this respect, the MSs in the rice genome are quite different from those in the human genome: in the latter, the majority of MSs show an obvious bias towards the high G+C content of DNA.Electronic Supplementary Material Supplementary material is available in the online version of this article at Communicated by M.-A. Grandbastien     

Variation in the expressivity of the ocular retardation gene in mice

Variation in the expressivity was studied of the gene for ocular retardation (or) in mice. It is shown that the gene or suppresses with a high expressivity the growth of the optic vesicle in homozygotes, this resulting in anophthalmia and microphthalmia with aphakia. In cases of low expressivity, the gene or inhibits the growth of retina anlage, this leading to microphthalmia with a cataract of the lens. Variation in the expressivity of the gene or is due to an influence of modifier genes.     

Times of crisis in the evolution of the cephalopoda

The evolutionary history of the ectocochlian cephalopods is punctuated by a number of severe crises during each of which this class came very close to extinction. The crisis events follow each other at intervals of from seven to almost 300 million years and, with one exception, were not synchronous for Nautiloidea and Ammonoidea. Only at the end of the Triassic period did both groups simultaneously face the danger of extinction. Generally, the survivors of crisis situations have simple shell forms and are strikingly similar to each other. To trace the details of cephalopod evolution, the family on the taxonomic level and the stratigraphic stage on the chronological level do not provide scales fine enough to reconstruct the true course of this process. The causes of crises and “mass extinctions” are not yet understood. Most authors have approached this problem in a simplistic manner, searching for a single cause for any, or all, events of this kind. It seems that we do not even have begun to understand what the problems are.     

On the evolution of saline tolerance in the larvae of mosquitoes in the genus Ochlerotatus

We examined physiological and morphological parameters associated with saline tolerance in the larvae of 11 species of mosquito in the genus Ochlerotatus that were collected from the wild in a variety of sites around North America. Saline tolerance was assayed, and all of the species were osmoregulators. Six of the species examined were found to be physiologically restricted to freshwater habitats, while the other five could successfully osmoregulate in both freshwater and saline water, including seawater. All larvae that were obligate freshwater forms had only one rectal segment, while all of the euryhaline osmoregulators had two. We were interested in the evolutionary pathway by which saline tolerance arose in this lineage. DNA sequence data were obtained by polymerase chain reaction amplification and sequencing of the D2 region of the 28s rDNA gene in all of the freshwater and saline-tolerant Ochlerotatus species we studied. When the morphological and physiological characters were mapped on the resultant cladogram, they revealed a complex pattern, with freshwater and saline-water forms being adjacent and interspersed through the tree. The data also demonstrate that saline tolerance has been gained and then lost at least once in this lineage. Two possible evolutionary scenarios are presented, but the one we favor is that saline tolerance arose one time in this lineage and repeated reversions to the freshwater condition have occurred.     

Dynamics of cypermethrin resistance in the field in the horn fly,Haematobia irritans

The toxicity of cypermethrin to the horn fly Haematobia irritans (L.) (Diptera: Muscidae) was determined for samples collected from untreated herds at a farm in central Argentina from October 1997 to May 2001. Field tests of the efficacy of cypermethrin against horn flies were first carried out at this farm in 1993, when the fly was shown to be susceptible to pyrethroids. Subsequently the horn fly populations on this farm were shown to have become resistant and, since 1997, the use of cypermethrin has been restricted to experimental purposes. In this study, fly samples collected in 1999, 2000 and 2001 were subjected to a polymerase chain reaction (PCR) to detect the presence of a specific nucleotide substitution in the sodium channel gene sequence, which has been associated with target site insensitivity to pyrethroids. This analysis showed that the level of cypermethrin resistance had diminished between 1997 and 2001. However, this was not sufficient to restore the efficacy of this pyrethroid to the level found prior to the onset of resistance. Heterozygous and homozygous resistant flies were detected in all samples of flies subjected to PCR diagnosis of alleles conferring target site resistance.     

Prospects for the involvement of cancer stem cells in the pathogenesis of osteosarcoma

Osteosarcoma (OS) is one of the most common bone tumors in children and adolescents that cause a high rate of mortality in this age group and tends to be metastatic, in spite of chemotherapy and surgery. The main reason for this can be returned to a small group of malignant cells called cancer stem cells (CSCs). OS-CSCs play a key role in the resistance to treatment and relapse and metastasis through self-renewal and differentiation abilities. In this review, we intend to go through the different aspects of this malignant disease, including the cancer stem cell-phenotype, methods for isolating CSCs, signaling pathways, and molecular markers in this disease, and drugs showing resistance in treatment efforts of OS.     

Endopeptidase-24.11 in the adenohypophysis of the pig is localized in the gonadotrophic cells

Endopeptidase-24.11, an ectoenzyme with a key role in metabolizing peptides at cell surfaces, is present in the adenohypophysis. A specific polyclonal antibody to the endopeptidase has been used to explore its localization in cryostat sections of pig pituitary glands by an immunoperoxidase method. Immunoreactivity was symmetrically but not uniformly distributed over the anterior lobe, with the highest intensity a zone just beneath the capsule along the anterior surface. In detail, the staining was observed to be in the cell membrane, but in some cells a small area of intense paranuclear staining was also observed. Serial 5 micron sections were immunostained alternately for endopeptidase-24.11 and for pituitary proteohormone. Luteinizing hormone (LH), follicular stimulating hormone (FSH), thyrotropin, adrenocorticotropin, prolactin and growth hormone were studied in this way. It was possible to identify groups of cells in adjacent sections and a good correlation was observed for endopeptidase-24.11-immunoreactivity with that for LH and FSH. The association of the endopeptidase with gonadotrophs was confirmed by double labelling. No evidence of colocalization was observed with the other proteohormone antibodies. We conclude that among the cells of the adenohypophysis only the gonadotrophs express endopeptidase-24.11 and discuss the possible significance of this observation in regard to the termination of peptide signals, such as that of luteinizing hormone-releasing hormone (LHRH) acting at this site.     

On the participation of RecB in the induction of mini-Tn10 precise excision in a dnaB thermosensitive mutant

Precise excision of transposons Tn10 and mini-Tn10 is increased in the dnaB252 thermosensitive mutant of Escherichia coli K12, at the permissive temperature. DNA repair proteins like Pol II, RecF, Ruv and RecA were found to participate, to different extents, in this induced excision event. In this work we report that DNA repair-recombination protein RecBCD has a predominant role in this deletion process. The role of this and other repair proteins in DNA replication of the dnaB mutant in relation to the excision of the transposon is analyzed.     

Participation of the liver gluconeogenesis in the glibenclamide-induced hypoglycaemia in rats

We previously demonstrated an increased liver gluconeogenesis (LG) during insulin‐induced hypoglycaemia. Thus, an expected effect of sulphonylureas induced hypoglycaemia (SIH) could be the activation of LG. However, sulphonylureas infused directly in to the liver inhibits LG. Considering these opposite effects we investigated herein LG in rats submitted to SIH. For this purpose, 24 h fasted rats that received glibenclamide (10 mg kg^?1) were used (SIH group). Control group received oral saline. Glycaemia at 30, 60, 90, 120 and 150 min after oral administration of glibenclamide were evaluated. Since the lowest glycaemia was obtained 120 min after glibenclamide administration, this time was chosen to investigate LG in situ perfused livers. The gluconeogenesis from precursors that enters in this metabolic pathway before the mitochondrial step, i.e. L ‐alanine (5 mM), L ‐lactate (2 mM), pyruvate (5 mM) and L ‐glutamine were decreased (p < 0·05). However, the gluconeogenic activity using glycerol (2 mM), which enters in the gluconeogenesis after the mitochondrial step was maintained. Taken together, the results suggest that the inhibition of LG promoted by SIH overcome the activation of this metabolic pathway promoted by IIH and could be attributed, at least in part, to its effect on mitochondrial function. Copyright © 2011 John Wiley & Sons, Ltd.     

Diversity of Phytases in the Rumen

Examples of a new class of phytase related to protein tyrosine phosphatases (PTP) were recently isolated from several anaerobic bacteria from the rumen of cattle. In this study, the diversity of PTP-like phytase gene sequences in the rumen was surveyed by using the polymerase chain reaction (PCR). Two sets of degenerate primers were used to amplify sequences from rumen fluid total community DNA and genomic DNA from nine bacterial isolates. Four novel PTP-like phytase sequences were retrieved from rumen fluid, whereas all nine of the anaerobic bacterial isolates investigated in this work contained PTP-like phytase sequences. One isolate, Selenomonas lacticifex, contained two distinct PTP-like phytase sequences, suggesting that multiple phytate hydrolyzing enzymes are present in this bacterium. The degenerate primer and PCR conditions described here, as well as novel sequences obtained in this study, will provide a valuable resource for future studies on this new class of phytase. The observed diversity of microbial phytases in the rumen may account for the ability of ruminants to derive a significant proportion of their phosphorus requirements from phytate.     

Problems in the Psychology of Activity

My purpose today is to sum up a number of the postulates concerning activity as it is used in psychology, without introducing any new ones; and I shall do this with the following in mind. Until now I have used a system of concepts I proposed earlier with respect to an analysis of activity; of course, I should like to work out a position, first and foremost my own, on this system, and to reexamine it. On the other hand, I should like to pose a number of questions, such as: Is this system of concepts of any significance, i.e., is it able to work in psychology? Obviously, this system must be developed, which essentially has not been done in recent years. This system of concepts is frozen, without any movement. I personally was quite alone in this respect. All movement takes place with regard to various problems contiguous with the problem of activity, some more, some less; but in my opinion, the concept of activity has been developed highly unsatisfactorily. That is why I thought I should try, today, to sum up very briefly what I think is important.     

Localization of the Outer Membrane Protein OmpA2 in Caulobacter crescentus Depends on the Position of the Gene in the Chromosome

The outer membrane of Gram-negative bacteria is an essential structure involved in nutrient uptake, protection against harmful substances, and cell growth. Different proteins keep the outer membrane from blebbing out by simultaneously interacting with it and with the cell wall. These proteins have been mainly studied in enterobacteria, where OmpA and the Braun and Pal lipoproteins stabilize the outer membrane. Some degree of functional redundancy exists between these proteins, since none of them is essential but the absence of two of them results in a severe phenotype. Caulobacter crescentus has a different strategy to maintain its outer membrane, since it lacks the Braun lipoprotein and Pal is essential. In this work, we characterized OmpA2, an OmpA-like protein, in this bacterium. Our results showed that this protein is required for normal stalk growth and that it plays a minor role in the stability of the outer membrane. An OmpA2 fluorescent fusion protein showed that the concentration of this protein decreases from the stalk to the new pole. This localization pattern is important for its function, and it depends on the position of the gene locus in the chromosome and, as a consequence, in the cell. This result suggests that little diffusion occurs from the moment that the gene is transcribed until the mature protein attaches to the cell wall in the periplasm. This mechanism reveals the integration of different levels of information from protein function down to genome arrangement that allows the cell to self-organize.     

Early steps in the development of the forebrain

The tremendous complexity of the adult forebrain makes it a challenging task to elucidate how this structure forms during embryonic development. Nevertheless, we are beginning to understand how a simple epithelial sheet of ectoderm gives rise to the labyrinthine network of cells that constitutes the functional forebrain. Here, we discuss early events in forebrain development--those that lead to the establishment of the anterior neural plate and the regional subdivision of this territory into the different domains of the prospective forebrain.     

Analysis of the DDE Motif in the Mutator Superfamily

The eukaryotic Mutator family of transposable elements is widespread in plants. Active or potentially active copies are also found in fungi and protozoans, and sequences related to this family have been detected in metazoans as well. Members of this family are called Mutator-like elements (MULE s). They encode transposases, which contain a region conserved with transposases of the IS256 prokaryotic family, known to harbor a DDE catalytic domain. Different DDE or D34E motifs have been proposed in some groups of eukaryotic MULEs based on primary sequence conservation. On a large number of protein sequences related to, and representative of, all MULE families, we analyzed global conservation, the close environment of different acidic residues and the secondary structure. This allowed us to identify a potential DDE motif that is likely to be homologous to the one in IS256-like transposases. The characteristics of this motif are depicted in each known family of MULEs. Different hypotheses about the evolution of this triad are discussed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.