The Δ4-desaturation pathway for DHA biosynthesis is operative in the human species: Differences between normal controls and children with the Zellweger syndrome

<p>Abstract</p> <p>Background</p> <p>Docosahexaenoic acid (DHA, 22:6ω3) is a fundamental component of cell membranes, especially in the brain and retina. In the experimental animal, DHA deficiency leads to suboptimal neurological performance and visual deficiencies. Children with the Zellweger syndrome (ZS) have a profound DHA deficiency and symptoms that can be attributed to their extremely low DHA levels. These children seem to have a metabolic defect in DHA biosynthesis, which has never been totally elucidated. Treatment with DHA ethyl ester greatly improves these patients, but if we could normalize their endogenous DHA production we could get additional benefits. We examined whether DHA biosynthesis by Δ4-desaturation could be enhanced in the human species by transfecting the enzyme, and if this could normalize the DHA levels in cells from ZS patients.</p> <p>Results</p> <p>We showed that the Δ4-desaturase gene (<it>Fad4</it>) from <it>Thraustochytrium sp</it>, which can be expressed by heterologous transfection in other plant and yeast cells, can also be transfected into human lymphocytes, and that it expresses the enzyme (FAD4, Δ4-desaturase) by producing DHA from direct Δ4-desaturation of 22:5ω3. We also found that the other substrate for Δ4-desaturase, 22:4ω6, was parallely desaturated to 22:5ω6.</p> <p>Conclusions</p> <p>The present "in vitro" study demonstrates that Δ4-desaturase can be transfected into human cells and synthesize DHA (as well as 22:5ω6, DPA) from 22:5ω3 and 22:4ω6, respectively, by putative Δ4-desaturation. Even if this pathway may not be the physiological route for DHA biosynthesis "in vivo", the present study opens new perspectives for the treatment of patients within the ZS spectrum.</p

UTP affects the Schwannoma cell line proteome through P2Y receptors leading to cytoskeletal reorganisation

Glial cells in the peripheral nervous system, such as Schwann cells, respond to nucleotides, which play an important role in axonal regeneration and myelination. Metabotropic P2Y receptor agonists are promising therapeutic molecules for peripheral neuropathies. Nevertheless, the proteomic mechanisms involved in nucleotide action on Schwann cells remain unknown. Here, we studied intracellular protein changes in RT4-D6P2T Schwann cells after treatment with nucleotides and Nucleo CMP Forte (CMPF), a nucleotide-based drug. After treatment with CMPF, 2-D DIGE revealed 11 differential gel spots, which were all upregulated. Among these, six different proteins were identified by MS. Some of these proteins are involved in actin remodelling (actin-related protein, Arp3), membrane vesicle transport (Rab GDP dissociation inhibitor β, Rab GDI), and the endoplasmic reticulum stress response (protein disulfide isomerase A3, PDI), which are hallmarks of a possible P2Y receptor signalling pathway. Expression of P2Y receptors in RT4-D6P2T cells was demonstrated by RT-PCR and a transient elevation of intracellular calcium measured in response to UTP. Actin reorganisation was visualized after UTP treatment using phalloidin-FITC staining and was blocked by the P2Y antagonist suramin, which also inhibited Arp3, Rab GDI, and PDI protein upregulation. Our data indicate that extracellular UTP interacts with Schwann P2Y receptors and activates molecular machinery that induces changes in the glial cell cytoskeleton. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.This research was supported by an unrestricted research grant from Ferrer S.A. (Barcelona).Peer Reviewe

Is Ankyrin a specific genetic risk factor for psychiatric phenotypes?

Background: Genome wide association studies reported two single nucleotide polymorphisms in ANK3 (rs9804190 and rs10994336) as independent genetic risk factors for bipolar disorder. Another SNP in ANK3 (rs10761482) was associated with schizophrenia in a large European sample. Within the debate on common susceptibility genes for schizophrenia and bipolar disorder, we tried to investigate common findings by analyzing association of ANK3 with schizophrenia, bipolar disorder and unipolar depression. Methods: We genotyped three single nucleotide polymorphisms (SNPs) in ANK3 (rs9804190, rs10994336, and rs10761482) in a case-control sample of German descent including 920 patients with schizophrenia, 400 with bipolar affective disorder, 220 patients with unipolar depression according to ICD 10 and 480 healthy controls. Sample was further differentiated according to Leonhard’s classification featuring disease entities with specific combination of bipolar and psychotic syndromes. Results: We found no association of rs9804190 and rs10994336 with bipolar disorder, unipolar depression or schizophrenia. In contrast to previous findings rs10761482 was associated with bipolar disorder (p = 0.015) but not with schizophrenia or unipolar depression. We observed no association with disease entities according to Leonhard’s classification. Conclusion: Our results support a specific genetic contribution of ANK3 to bipolar disorder though we failed to replicate findings for schizophrenia. We cannot confirm ANK3 as a common risk factor for different diseases

Assessment of greenhouse emissions of the green bean through the static enclosure technique

Unidad de excelencia María de Maeztu CEX2019-000940-MUrban green installations are extensively promoted to increase sustainable and accessible food production and simultaneously improve the environmental performance and liveability of city buildings. In addition to the multiple benefits of plant retrofitting, these installations may lead to a consistent increase in biogenic volatile organic compounds (BVOCs) in the urban environment, especially indoors. Accordingly, health concerns could limit the implementation of building-integrated agriculture. In a building-integrated rooftop greenhouse (i-RTG), throughout the whole hydroponic cycle, green bean emissions were dynamically collected in a static enclosure. Four representative BVOCs, α-pinene (monoterpene), β-caryophyllene (sesquiterpene), linalool (oxygenated monoterpene) and cis-3-hexenol (LOX derivate), were investigated in the samples collected from two equivalent sections of a static enclosure, one empty and one occupied by the i-RTG plants, to estimate the volatile emission factor (EF). Throughout the season, extremely variable BVOC levels between 0.04 and 5.36 ppb were found with occasional but not significant (P > 0.05) variations between the two sections. The highest emission rates were observed during plant vegetative development, with EFs equivalent to 78.97, 75.85 and 51.34 ng g−1 h−1 for cis-3-hexenol, α-pinene, and linalool, respectively; at plant maturity, all volatiles were either close to the LLOQ (lowest limit of quantitation) or not detected. Consistent with previous studies significant relationships (r ≥ 0.92; P < 0.05) were individuated within volatiles and temperature and relative humidity of the sections. However, correlations were all negative and were mainly attributed to the relevant effect of the enclosure on the final sampling conditions. Overall, levels found were at least 15 folds lower than the given Risk and LCI values of the EU-LCI protocol for indoor environments, suggesting low BVOC exposure in the i-RTG. Statistical outcomes demonstrated the applicability of the static enclosure technique for fast BVOC emissions survey inside green retrofitted spaces. However, providing high sampling performance over entire BVOCs collection is recommended to reduce sampling error and incorrect estimation of the emissions

Can enzyme kinetics of prooxidants teach us a lesson about the treatment of Alzheimer's disease: a pilot post-mortem study

OBJECTIVES: Oxidative stress (OS), is defined as an imbalance of pro- and antioxidants, leading to increased production of free radicals, which can lead to cell damage and death, has been postulated as important factors in the pathogenesis of neurodegenerative disorders such as Alzheimer's disease (AD). Most research has concentrated on the antioxidant system, for the first time, this proof of concept study examines the prooxidant system by investigating kinetic parameters of the free radical producing enzyme xanthine oxidase directly in post mortem brain tissue. METHODS: We determined the Michaelis-Menten constant (K(M)) and the maximal velocity (V(Max)) of xanthine oxidase (XO) in the cortico-limbic system of patients with AD using activity assays. RESULTS: We found the Michaelis-Menton constant of XO significantly decreased in hippocampus of patients with AD compared to controls. None of the other brain regions showed any significant alterations of these parameters. CONCLUSIONS: These results add further evidence to the amount of research indicating that OS plays an important role in AD. Moreover, these results should encourage more research in this field and it maybe speculated that this might open new avenues for treatment and prevention in AD

Schizophrenia: from the brain to peripheral markers. A consensus paper of the WFSBP task force on biological markers

Objective. The phenotypic complexity, together with the multifarious nature of the so-called "schizophrenic psychoses", limits our ability to form a simple and logical biologically based hypothesis for the disease group. Biological markers are defined as biochemical, physiological or anatomical traits that are specific to particular conditions. An important aim of biomarker discovery is the detection of disease correlates that can be used as diagnostic tools. Method. A selective review of the WFSBP Task Force on Biological Markers in schizophrenia is provided from the central nervous system to phenotypes, functional brain systems, chromosomal loci with potential genetic markers to the peripheral systems. Results. A number of biological measures have been proposed to be correlated with schizophrenia. At present, not a single biological trait in schizophrenia is available which achieves sufficient specificity, selectivity and is based on causal pathology and predictive validity to be recommended as diagnostic marker. Conclusions. With the emergence of new technologies and rigorous phenotypic subclassification the identification of genetic bases and assessment of dynamic disease related alterations will hopefully come to a new stage in the complex field of psychiatric research