Palate vault morphology in Down syndrome

Moire topography was used to quantitatively determine the shape of the palatal vault in 57 Spanish patients with Down syndrome (DS) (38 males and 19 females; age range 18-36 years) and in 100 normal controls (N) (76 males and 24 females; age range 20-29 years). The topographic image for each palatal vault was analyzed and approximately 40 sets of coordinates (x, y, z) were recorded. Other parameters, including length, width and maximum height, were recorded from the palate vault images. No appreciable sexual dimorphism in palate dimensions was observed in DS subjects versus the situation in N subjects. Globally, the average N dimensions were significantly greater than those in the DS patients (P < 0.005). A special palatal morphology was found to be associated with DS, with different ratios for the three dimensions (mean factor 0.88 for length, 0.81 for width and 0.73 for height), versus the healthy palatal vaults. It is concluded that palatal morphology in DS fits an elliptic paraboloid. On the other hand, no lineal correlation was observed between height, width and length in the DS and N groups. The scatter plots of bivariate data exhibited a shapeless morphology. The lineal correlation coefficients ranged from 0.008 to 0.33 for the DS and N groups.     

Nitrogen availability influences the biochemical composition and photosynthesis of tank-cultivated Ulva rigida (Chlorophyta)

Physiological and biochemical changes in relation to inorganic nitrogen availability were studied for tank-cultivated Ulva rigida grown under nitrogen- enriched and nitrogen-depleted seawater. U. rigida was initially cultivated in nitrogen-enriched seawater (daily concentrations of NH4+ and NO3- + NO2- ranged between 0.5–1.7 and 0.06–0.15 mg L-1, respectively), then transferred to nitrogen-depleted seawater where photosynthetic capacity decreased to zero after 23 d. At the time (14 d) when photosynthetic rates were lower than 2.0 μmol O2 g-1 FW min-1 and strong bleaching had occurred, some algae were returned to the initial nitrogen-enriched seawater to study recovery from N-limited growth. Data on biochemical composition (chlorophylls, ash, caloric content, fatty acids and dietary fibres) and colouration varied significantly depending on the nitrogen conditions. C:N ratios correlated significantly with biochemical parameters. Fatty acid (FA) synthesis continued during the N-starvation period; saturated and mono-unsaturated FA increased to a maximun of 72.2%, while poly-unsaturated fatty acids (PUFA) decreased to 27.7%. During the N-enriched recovery period, the reverse was found. C:N ratios above 10 correlated with carbohydrate synthesis as shown by the dietary fibre level. Under nitrogen enriched conditions, C:N ratios decreased along with a decrease in fibre level. Under controlled conditions, nitrogen represents a major influence on the development of intensive tank cultivation of Ulva rigida, not only by affecting parameters closely related to nitrogen metabolism but also some clearly influenced by carbon uptake. This revised version was published online in June 2006 with corrections to the Cover Date.     

Quality Control Test for Sequence-Phenotype Assignments

Relating a gene mutation to a phenotype is a common task in different disciplines such as protein biochemistry. In this endeavour, it is common to find false relationships arising from mutations introduced by cells that may be depurated using a phenotypic assay; yet, such phenotypic assays may introduce additional false relationships arising from experimental errors. Here we introduce the use of high-throughput DNA sequencers and statistical analysis aimed to identify incorrect DNA sequence-phenotype assignments and observed that 10–20% of these false assignments are expected in large screenings aimed to identify critical residues for protein function. We further show that this level of incorrect DNA sequence-phenotype assignments may significantly alter our understanding about the structure-function relationship of proteins. We have made available an implementation of our method at http://bis.ifc.unam.mx/en/software/chispas.     

Dissecting drug and vehicle metabolic effects in rats by a metabonomic approach

A combined application of high resolution (1)H NMR spectroscopy and multivariate statistical techniques focused on establishing a consistent statistical approach to metabonomic studies was tested. The data reduction, which is preliminary to the application of multivariate analysis to NMR spectra, was carried out by means of two complementary methods: pure Pattern Recognition (PR) and Assigned Signal Analysis (ASA). The simultaneous use of both approaches allowed us to obtain additional information in the analysis of metabonomic data, compared to the use of PR alone. This additional information consists in the possibility of a biochemical interpretation of the effects induced by treatment with xenobiotics, such as drugs or drug vehicles, on the metabolic networks of the systems under investigation. This approach allowed us to ascertain that a single-dose treatment with ST1959 vehicled by Sesame oil affects the production of hepatic glucose associated to an increment of the amino acid ketogenic process.     

Outdoor cultivation of microalgae for carotenoid production: current state and perspectives

Microalgae are a major natural source for a vast array of valuable compounds, including a diversity of pigments, for which these photosynthetic microorganisms represent an almost exclusive biological resource. Yellow, orange, and red carotenoids have an industrial use in food products and cosmetics as vitamin supplements and health food products and as feed additives for poultry, livestock, fish, and crustaceans. The growing worldwide market value of carotenoids is projected to reach over US$1,000 million by the end of the decade. The nutraceutical boom has also integrated carotenoids mainly on the claim of their proven antioxidant properties. Recently established benefits in human health open new uses for some carotenoids, especially lutein, an effective agent for the prevention and treatment of a variety of degenerative diseases. Consumers’ demand for natural products favors development of pigments from biological sources, thus increasing opportunities for microalgae. The biotechnology of microalgae has gained considerable progress and relevance in recent decades, with carotenoid production representing one of its most successful domains. In this paper, we review the most relevant features of microalgal biotechnology related to the production of different carotenoids outdoors, with a main focus on β-carotene from Dunaliella, astaxanthin from Haematococcus, and lutein from chlorophycean strains. We compare the current state of the corresponding production technologies, based on either open-pond systems or closed photobioreactors. The potential of scientific and technological advances for improvements in yield and reduction in production costs for carotenoids from microalgae is also discussed.     

Outdoor cultivation of lutein-rich cells of <Emphasis Type="Italic">Muriellopsis</Emphasis> sp. in open ponds

The growth performance of the chlorophycean microalga Muriellopsis sp. outdoors in open tanks agitated with a paddlewheel and its ability to accumulate carotenoids have been evaluated throughout the year. The cells grown in the open system had free lutein as the main carotenoid, with violaxanthin, β-carotene, and neoxanthin also present. Lutein content of the dry biomass ranged from 0.4 to 0.6%, depending on the growth and environmental conditions. In addition, the biomass of Muriellopsis sp. had a high content in both protein and lipids with about half of the fatty acids being of the polyunsaturated type, with α-linolenic acid accounting for almost 30% of the total fatty acids. The effect of determinant parameters on the performance of the cultures in open tanks was evaluated. Operating conditions that allow the maintenance of productive cultures were established under semicontinuous regime for 9 months throughout the year. Biomass and lutein yields in the open system were not far from those in closed tubular photobioreactors, and reached productivity values of 20 g dry biomass, containing around 100 mg lutein m⁻² day⁻¹ in summer. The outdoor culture of Muriellopsis sp. in open ponds thus represents a real alternative to established systems for the production of lutein.     

Involvement of DEAD-box proteins in group I and group II intron splicing. Biochemical characterization of Mss116p, ATP hydrolysis-dependent and -independent mechanisms, and general RNA chaperone activity

The RNA-catalyzed splicing of group I and group II introns is facilitated by proteins that stabilize the active RNA structure or act as RNA chaperones to disrupt stable inactive structures that are kinetic traps in RNA folding. In Neurospora crassa and Saccharomyces cerevisiae, the latter function is fulfilled by specific DEAD-box proteins, denoted CYT-19 and Mss116p, respectively. Previous studies showed that purified CYT-19 stimulates the in vitro splicing of structurally diverse group I and group II introns, and uses the energy of ATP binding or hydrolysis to resolve kinetic traps. Here, we purified Mss116p and show that it has RNA-dependent ATPase activity, unwinds RNA duplexes in a non-polar fashion, and promotes ATP-independent strand-annealing. Further, we show that Mss116p binds RNA non-specifically and promotes in vitro splicing of both group I and group II intron RNAs, as well as RNA cleavage by the aI5gamma-derived D135 ribozyme. However, Mss116p also has ATP hydrolysis-independent effects on some of these reactions, which are not shared by CYT-19 and may reflect differences in its RNA-binding properties. We also show that a non-mitochondrial DEAD-box protein, yeast Ded1p, can function almost as efficiently as CYT-19 and Mss116p in splicing the yeast aI5gamma group II intron and less efficiently in splicing the bI1 group II intron. Together, our results show that Mss116p, like CYT-19, can act broadly as an RNA chaperone to stimulate the splicing of diverse group I and group II introns, and that Ded1p also has an RNA chaperone activity that can be assayed by its effect on splicing mitochondrial introns. Nevertheless, these DEAD-box protein RNA chaperones are not completely interchangeable and appear to function in somewhat different ways, using biochemical activities that have likely been tuned by coevolution to function optimally on specific RNA substrates.     

A fluorescence assay for peptide translocation into mitochondria

Translocation of the presequence is an early event in import of preproteins across the mitochondrial inner membrane by the TIM23 complex. Import of signal peptides, whose sequences mimic mitochondrial import presequences, was measured using a novel, qualitative, fluorescence assay in about 1h. This peptide assay was used in conjunction with classical protein import analyses and electrophysiological approaches to examine the mechanisms underlying the functional effects of depleting two TIM23 complex components. Tim23p forms, at least in part, the pore of this complex while Tim44p forms part of the translocation motor. Depletion of Tim23p eliminates TIM23 channel activity, which interferes with both peptide and preprotein translocation. In contrast, depletion of Tim44p disrupts preprotein but not peptide translocation, which has no effect on TIM23 channel activity. Two conclusions were made. First, this fluorescence peptide assay was validated as two different mutants were accurately identified. Hence, this assay could provide a rapid means of screening mutants to identify those that fail an initial step in import, i.e., translocation of the presequence. Second, translocation of signal peptides required normal channel activity and disruption of the presequence translocase-associated motor complex did not modify TIM23 channel activity nor prevent presequence translocation.