An Untargeted Metabolomics Approach to Characterize Short-Term and Long-Term Metabolic Changes after Bariatric Surgery.

Bariatric surgery is currently one of the most effective treatments for obesity and leads to significant weight reduction, improved cardiovascular risk factors and overall survival in treated patients. To date, most studies focused on short-term effects of bariatric surgery on the metabolic profile and found high variation in the individual responses to surgery. The aim of this study was to identify relevant metabolic changes not only shortly after bariatric surgery (Roux-en-Y gastric bypass) but also up to one year after the intervention by using untargeted metabolomics.

Surgical management of cross-bites in orthognathic surgery: Surgically assisted rapid maxillary expansion (SARME) versus two-piece maxilla.

Purpose: The surgical treatment of cross-bites includes surgically-assisted maxillary expansion (SARME) or maxillary-bipartition during bimaxillary surgery. This study evaluates and compares the changes in the teeth and lower nasal passage, as well as the stability of the expansion.

Patients And Methods: The measurements were performed on the cone-beam computed tomography (CBCT) scans of 32 patients with transverse (width) deficiencies of the maxilla.

IPO: a tool for automated optimization of XCMS parameters.

Background: Untargeted metabolomics generates a huge amount of data. Software packages for automated data processing are crucial to successfully process these data. A variety of such software packages exist, but the outcome of data processing strongly depends on algorithm parameter settings.

Enhanced doxorubicin delivery to the brain administered through glutathione PEGylated liposomal doxorubicin (2B3-101) as compared with generic Caelyx,(®)/Doxil(®)--a cerebral open flow microperfusion pilot study.

The neuroprotective blood-brain barrier (BBB) keeps many drug candidates below therapeutic levels in the central nervous system. Glutathione PEGylated liposomal doxorubicin (2B3-101) has been developed to safely enhance the delivery of doxorubicin to brain tumors. However, doxorubicin concentration in extracellular brain fluid cannot yet be reliably measured using conventional techniques.

Polyamines in biological samples: rapid and robust quantification by solid-phase extraction online-coupled to liquid chromatography-tandem mass spectrometry.

Polyamines are ubiquitous active biogenic amines which contribute to basic cellular functions. Hence, their quantification in samples of diverse biological origins is essential for understanding how they function, especially in disease-relevant conditions. We present here a robust, high-throughput solid-phase extraction online coupled to a liquid chromatography-tandem mass spectrometry (SPE-LC/MS/MS) approach for the simultaneous quantification of eight polyamines in various biological samples.

Effect of ceramide on nonraft proteins.

The currently accepted model of biological membranes involves a heterogeneous, highly dynamic organization, where certain lipids and proteins associate to form cooperative platforms ("rafts") for cellular signaling or transport processes. Ceramides, a lipid species occurring under conditions of cellular stress and apoptosis, are considered to stabilize these platforms, thus modulating cellular function. The present study focuses on a previously unrecognized effect of ceramide generation.

A cDNA sequence coding for a glutamic acid-rich protein is differentially expressed in cassava storage roots.

We report the isolation and characterization of a cDNA sequence (Mec1) coding for a glutamic acid-rich protein (Pt2L4) from cassava storage roots. Comparative sequence analysis showed a high identity of Pt2L4 with cassava protein C54, which is expressed in vascular tissues of storage roots. Northern blot analysis showed that the Mec1 transcript expression pattern might be related to the maturation of the storage parenchyma cells.

An opaque-2-like transcription factor from pearl millet.

Pearl millet (Pennisetum glaucum L.), a species of the Poaceae family, is an important food crop in Africa, Asia and South America. Its nutritional value is due to storage prolamins accumulated in the seeds.

Towards the identification of cassava root protein genes.

The protein population of cassava root layers was characterized by SDS-PAGE and bidimensional polyacrylamide gel electrophoresis. SDS-Page revealed the presence of a protein population in the molecular weight range between 94 and 20 kDa. The expression pattern of these proteins was well-defined within the different layers.

Characterization of pearl millet prolamins.

We report the physical-chemical characterization of the major alcohol-soluble proteins present in seeds of pearl millet (Pennisetum glaucum) by SDS-PAGE, bidimensional gel electrophoresis, MALDI-TOF/MS and RP-HPLC. We demonstrate the presence of three major prolamins, called A-, B- and C-pennisetin with mass values around 27, 22 and 12 kDa, respectively. We present partial amino acid sequences of these major proteins, which should allow the posterior isolation of the respective genes.

ACGT and vicilin core sequences in a promoter domain required for seed-specific expression of a 2S storage protein gene are recognized by the opaque-2 regulatory protein.

The expression of Brazil nut storage albumin genes is highly regulated during seed development. Several sequences in the promoter of one of these genes show homologies with the target sites of the maize O2 bZIP regulatory protein. We therefore asked whether the O2 protein would recognize these promoter sequences.

Genome DNA sequencing around the EF-1 alpha multigene locus of Arabidopsis thaliana indicates a high gene density and a shuffling of noncoding regions.

In Arabidopsis thaliana, EF-1 alpha proteins are encoded by a multigene family of four members. Three of them are clustered at the same locus, which was positioned 24 cM from the top of chromosome 1. A region of DNA spanning 63 kb around these locus was sequenced and analyzed.

Modified 2S albumins with improved tryptophan content are correctly expressed in transgenic tobacco plants.

Brazil nut 2S albumins lack the essential amino acid tryptophan. In order to improve the protein's nutritional value and create a basis for structural investigations, three separate modified Brazil nut 2S albumin genes were constructed. The first mutant contains five consecutive tryptophan codons, while the other two modified genes encode proteins carrying single tryptophan residues at sites that will allow confirmation of the predicted protein structure through fluorescence quenching techniques.

A rapid and sensitive dot-blot hybridization assay for the detection of citrus exocortis viroid in Citrus medica with digoxigenin-labelled RNA probes.

A rapid and sensitive dot-blot hybridization assay using in vitro-transcribed digoxigenin-labelled RNA probes (riboprobes) was developed aiming at detection of citrus exocortis viroid (CEVd) in crude sap of infected Citrus medica plants. The protocol includes a very quick and simple preparation of RNA extracts from samples using a denaturation step with formaldehyde. From our results, the employment of this step is highly recommended because the hybridization signals in formaldehyde-denatured samples were significantly stronger when compared with that of extracts without formaldehyde treatment.

Transgenic plants of ramie (Boehmeria nivea Gaud.) obtained by Agrobacterium mediated transformation.

A regeneration and transformation protocol for ramie (Boehmeria nivea Gaud.) is presented. Regeneration was obtained from leaf discs placed on solid B-5 medium (Gamborg et al.

Particle bombardment-mediated transient expression of a Brazil nut methionine-rich albumin in bean (Phaseolus vulgaris L.).

Bean (Phaseolus vulgaris L.) mature embryos were transformed using biolistic methods with a plasmid containing 2S albumin and beta-glucuronidase structural sequences, both under the control of the 35S CaMV promoter. We have shown that chimaeric tissues could be obtained and that both structural sequences were expressed to similar levels.

Cis and trans-acting elements involved in the activation of Arabidopsis thaliana A1 gene encoding the translation elongation factor EF-1 alpha.

In A. thaliana the translation elongation factor EF-1 alpha is encoded by a small multigenic family of four members (A1-A4). The A1 gene promoter has been dissected and examined in a transient expression system using the GUS reporter gene.

Isolation, characterization and expression of a gene coding for a 2S albumin from Bertholletia excelsa (Brazil nut).

Two genes, BE2S1 and BE2S2, coding for methionine-rich albumins of Brazil nut (Bertholletia excelsa H.B.K.

Expression of the 2S albumin from Bertholletia excelsa in Brassica napus.

The methionine rich 2S albumin seed storage protein of Bertholletia excelsa has been expressed in seeds of Brassica napus (rapeseed). A chimeric gene driven by the soybean lectin 5' flanking regions was used to produce a fusion protein consisting of the soybean lectin signal peptide and the propeptide of the Brazil nut 2S albumin. Several transgenic plants were studied at the RNA and protein levels; in each case the chimeric gene was expressed and the protein detected at levels ranging from 0.

The compaction pattern of the chromatin of trypanosomes.

Protozoa of the family Trypanosomatidae are pathogenic agents of human and animal diseases. Fine structure, compaction pattern, and histone content of the soluble chromatin were investigated in procyclic forms of Trypanosoma cruzi (Chagas disease, S. America) and T.

Protein synthesis in purified trypo- and epimastigote forms of Trypanosoma cruzi.

In order to develop the experimental background for studies on the differentiation of the medically important lower eukaryote, Trypanosoma cruzi, the polypeptides synthesized by purified epimastigotes and trypomastigotes were examined. The in vivo synthesized proteins were compared with polypeptides synthesized in reticulocyte lysate systems programmed with total RNA of the two forms. Qualitative and quantitative differences between the protein populations of the two forms were detected.

Optimization of a protein synthesizing lysate system from Trypanosoma cruzi.

A protein-synthesizing lysate system from Trypanosoma cruzi epimastigotes analogous to the rabbit reticulocyte lysate system was established. The system was optimized by the 'classical' method where one of the factors is varied while the others are kept constant. With this the following optima were found: [Mg2+]: 1.

Morphologic and biochemical characterization of Crithidia brasiliensis sp. n.

A trypanosomatid with a choanomastigote stage and, therefore, belonging to the genus Crithidia, was isolated in culture from the alimentary tract of the hemipteran genus Zelus. The trypanosomatid was able to grow at 37 C, a characteristic reported to date from only 2 other members of Crithidia, C hutneri and C. luciliae thermophila.