Jorge A. Saraiva

Haploinsufficiency of TCF4 causes syndromal mental retardation with intermittent hyperventilation (Pitt-Hopkins syndrome).

Pitt-Hopkins syndrome is a rarely reported syndrome of so-far-unknown etiology characterized by mental retardation, wide mouth, and intermittent hyperventilation. By molecular karyotyping with GeneChip Human Mapping 100K SNP arrays, we detected a 1.2-Mb deletion on 18q21.2 in one patient. Sequencing of the TCF4 transcription factor gene, which is contained in the deletion region, in 30 patients with significant phenotypic overlap revealed heterozygous stop, splice, and missense mutations in five further patients with severe mental retardation and remarkable facial resemblance. Thus, we establish the Pitt-Hopkins syndrome as a distinct but probably heterogeneous entity caused by autosomal dominant de novo mutations in TCF4. Because of its phenotypic overlap, Pitt-Hopkins syndrome evolves as an important differential diagnosis to Angelman and Rett syndromes. Both null and missense mutations impaired the interaction of TCF4 with ASCL1 from the PHOX-RET pathway in transactivating an E box-containing reporter construct; therefore, hyperventilation and Hirschsprung disease in patients with Pitt-Hopkins syndrome might be explained by altered development of noradrenergic derivatives.

Effect of the ionic liquid [bmim]Cl and high pressure on the activity of cellulase

The effect of the ionic liquid 1-butyl-3-methylimidazolium chloride ([bmim]Cl) and of high pressure on the activity of cellulase from Aspergillus niger were studied separately and in combination. The enzyme activity decreased with increasing concentrations of [bmim]Cl, reaching 50% the value in aqueous buffer with 20% [bmim]Cl. However, when the enzyme is held in 10% [bmim]Cl and is then assayed in 1% [bmim]Cl, it showed only 8% reduction of activity. These results can be explained by the fact that the activity of the enzyme in [bmim]Cl is linearly correlated with the decrease of the thermodynamic water activity (aw). Under pressure the enzyme activity varied from less 60% (at 200MPa) to equal (at 400 MPa), compared to atmospheric pressure. In 10% [bmim]Cl under pressure, cellulase activity is improved compared to atmospheric pressure, varying from equal (at 600 MPa) to 1.7-fold higher (at 100 MPa). This opens the possibility to improve cellulase activity in ionic liquids, and possibly of other enzymes, by carrying out the reaction under pressure.

Chemical composition and bioactivity of different oregano (Origanum vulgare) extracts and essential oil

BACKGROUND There is a growing interest in industry to replace synthetic chemicals by natural products with bioactive properties. Aromatic plants are excellent sources of bioactive compounds that can be extracted using several processes. As far as oregano is concerned, studies are lacking addressing the effect of extraction processes in bioactivity of extracts. This study aimed to characterise the in vitro antioxidant and antibacterial properties of oregano (Origanum vulgare) essential oil and extracts (in hot and cold water, and ethanol), and the chemical composition of its essential oil. RESULTS The major components of oregano essential oil were carvacrol, β-fenchyl alcohol, thymol, and γ-terpinene. Hot water extract had the strongest antioxidant properties and the highest phenolic content. All extracts were ineffective in inhibiting the growth of the seven tested bacteria. In contrast, the essential oil inhibited the growth of all bacteria, causing greater reductions on both Listeria strains (L. monocytogenes and L. innocua). CONCLUSION O. vulgare extracts and essential oil from Portuguese origin are strong candidates to replace synthetic chemicals used by the industry.

Chemical and physical methodologies for the replacement/reduction of sulfur dioxide use during winemaking: review of their potentialities and limitations

Sulfur dioxide (SO2) is probably one of the most versatile and efficient additives used in winemaking due to its antiseptic and antioxidant properties. This compound is also important for minimizing phenolic polymerization rate and color loss during wine aging. However, allergies caused by SO2-derived compounds, namely the sulfites, are becoming more frequent, causing symptoms such as headaches, nausea, gastric irritation, and breathing difficulties in asthma patients. Consequently, the legislated maximum concentration of SO2 allowed in wines has been gradually reduced. For this reason, it is crucial in a competitive global winemaking market strategy, to reduce or even eliminate the use of SO2 as a preservative and to search for new healthier and safe strategies. This work gives an overview of the main methodologies that have been proposed so far and that have potential to be used in winemaking as an alternative to SO2. The addition of compounds such as dimethyl dicarbonate, bacteriocins, phenolic compounds, and lysozyme, and the use of physical methods, namely pulsed electric fields, ultrasound, ultraviolet radiation, and high pressure are discussed and critically evaluated.

Microorganisms under high pressure — Adaptation, growth and biotechnological potential

Hydrostatic pressure is a well-known physical parameter which is now considered an important variable of life, since organisms have the ability to adapt to pressure changes, by the development of resistance against this variable. In the past decades a huge interest in high hydrostatic pressure (HHP) technology is increasingly emerging among food and biosciences researchers. Microbial specific stress responses to HHP are currently being investigated, through the evaluation of pressure effects on biomolecules, cell structure, metabolic behavior, growth and viability. The knowledge development in this field allows a better comprehension of pressure resistance mechanisms acquired at sub-lethal pressures. In addition, new applications of HHP could arise from these studies, particularly in what concerns to biotechnology. For instance, the modulation of microbial metabolic pathways, as a response to different pressure conditions, may lead to the production of novel compounds with potential biotechnological and industrial applications. Considering pressure as an extreme life condition, this review intends to present the main findings so far reported in the scientific literature, focusing on microorganisms with the ability to withstand and to grow in high pressure conditions, whether they have innated or acquired resistance, and show the potential of the application of HHP technology for microbial biotechnology.

Landmarks in the historical development of twenty first century food processing technologies

Over a course of centuries, various food processing technologies have been explored and implemented to provide safe, fresher-tasting and nutritive food products. Among these technologies, application of emerging food processes (e.g., cold plasma, pressurized fluids, pulsed electric fields, ohmic heating, radiofrequency electric fields, ultrasonics and megasonics, high hydrostatic pressure, high pressure homogenization, hyperbaric storage, and negative pressure cavitation extraction) have attracted much attention in the past decades. This is because, compared to their conventional counterparts, novel food processes allow a significant reduction in the overall processing times with savings in energy consumption, while ensuring food safety, and ample benefits for the industry. Noteworthily, industry and university teams have made extensive efforts for the development of novel technologies, with sound scientific knowledge of their effects on different food materials. The main objective of this review is to provide a historical account of the extensive efforts and inventions in the field of emerging food processing technologies since their inception to present day.

Concentration effect of hydrophilic ionic liquids on the enzymatic activity of Candida antarctica lipase B

A systematic study of the effects of hydrophilic ionic liquids concentration and nature (alkyl chain length and type of anion) on the activity of Candida antarctica lipase B is here reported. The increase in the concentration of the studied ionic liquids is shown to cause a decrease of the enzyme activity, but the effect is dependent on the ionic liquid used. This behavior is partially due to the ionic liquid impact on the thermodynamic water activity, but direct interactions between the hydrophilic ionic liquid and the enzyme are also disclosed. Cations with longer alkyl chains decrease the enzyme activity by obstruction of its non-polar active site, while direct interactions established between the enzyme and the anions, dominated by dispersion forces and hydrogen-bonding, contribute also for the loss of activity observed.

Ionic liquids microemulsions: the key to Candida antarctica lipase B superactivity

The activity of the commercial enzyme Candida antarctica lipase B (CaLB), in aqueous solutions of the ionic liquid 1-decyl-3-methylimidazolium chloride, [C10mim]Cl, at pH 7.0 was investigated. The relative enzyme activity (ActIL/ActBf) for ionic liquid molar concentrations ranging from 0.000–0.150 mol L−1 show an increase up to six fold in ActIL/ActBf, with the ionic liquid molar concentration. This phenomenon is shown to be related with the formation of micelles originated by the self-aggregation of [C10mim]Cl. The enzyme activation energy is not affected by the ionic liquid presence, suggesting the absence of structural changes in the enzyme induced by the ionic liquid. It is here demonstrated, for the first time, that it is possible to significantly increase the activity of an enzyme simply by using aqueous solutions of ionic liquids.

Visual phenotype in Williams-Beuren syndrome challenges magnocellular theories explaining human neurodevelopmental visual cortical disorders

Williams-Beuren syndrome (WBS), a neurodevelopmental genetic disorder whose manifestations include visuospatial impairment, provides a unique model to link genetically determined loss of neural cell populations at different levels of the nervous system with neural circuits and visual behavior. Given that several of the genes deleted in WBS are also involved in eye development and the differentiation of retinal layers, we examined the retinal phenotype in WBS patients and its functional relation to global motion perception. We discovered a low-level visual phenotype characterized by decreased retinal thickness, abnormal optic disk concavity, and impaired visual responses in WBS patients compared with age-matched controls by using electrophysiology, confocal and coherence in vivo imaging with cellular resolution, and psychophysics. These mechanisms of impairment are related to the magnocellular pathway, which is involved in the detection of temporal changes in the visual scene. Low-level magnocellular performance did not predict high-level deficits in the integration of motion and 3D information at higher levels, thereby demonstrating independent mechanisms of dysfunction in WBS that will require remediation strategies different from those used in other visuospatial disorders. These findings challenge neurodevelopmental theories that explain cortical deficits based on low-level magnocellular impairment, such as regarding dyslexia.

Effect of high pressure treatments on the physicochemical properties of a sulphur dioxide-free red wine.

The application of high hydrostatic pressure (HHP) in winemaking for substitution of the use of sulphur dioxide is still at a very early stage of development, since knowledge about the effect on physicochemical and sensorial characteristics of the wine during storage is very scarce. In this work, the evolution of colour, antioxidant activity and total phenolic compounds of SO2-free red wines treated by HHP and aged in bottles was followed for 12 months. The pressurised wines were compared with wine samples prepared with addition of 40 ppm of SO2 and without any of these two treatments. After 12 months, the pressurised wines presented higher values of CIELab parameters (a(*), b(*), and L(*)) and a lower monomeric anthocyanin content (45-61%) when compared to the unpressurised ones. The pressurised wines showed also a better global sensorial assessment, with the pressure treatments imparting aged-like characteristics to the wines. The wine deposits of pressurised wines had higher total phenolic content, namely proanthocyanidins (3- to 10-fold). The results demonstrate that HHP can influence long term red wine physicochemical and sensorial characteristics, hypothesised to be due to an increase of condensation reactions of phenolic compounds, forming compounds with higher degree of polymerisation that became insoluble in wine along storage.