Molecular analysis of PKU-associated PAH mutations: a fast and simple genotyping test

Abstract: Neonatal screening for phenylketonuria (PKU, OMIM: 261600) was introduced at the end of the 1960s. We developed a rapid and simple molecular test for the most frequent phenylalanine hydroxylase (PAH, Gene ID: 5053) mutations. Using this method to detect the 18 most frequent mutations, it is possible to achieve a 75% detection rate in Italian population. The variants selected also reach a high detection rate in other populations, for example, 70% in southern Germany, 68% in western Germany, 76% in Denmark, 68% in Sweden, 63% in Poland, and 60% in Bulgaria. We successfully applied this confirmation test in neonatal screening for hyperphenylalaninemias using dried blood spots and obtained the genotype in approximately 48 h. The method was found to be suitable as second tier test in neonatal screening for hyperphenylalaninemias in neonates with a positive screening test. This test can also be useful for carrier screening because it can bypass the entire coding sequence and intron–exon boundaries sequencing, thereby overcoming the questions that this approach implies, such as new variant interpretations

Use of Bone Marrow Aspirate Concentrate (BMAC) Associated with Hyperbaric Oxygenation Therapy in Maxillary Appositional Bone Reconstruction. A Randomized Clinical Trial

Objectives: The objective of this study was to evaluate bone reconstruction using xenograft alone and associated with bone marrow aspirate concentrate (BMAC) and hyperbaric oxygen therapy. Material and Methods: Twenty-four maxillary edentulous patients were randomly assigned into three groups: Control group (CG)—xenograft bone alone (n = 8); Group 1 (G1)—xenogeneic bone block combined with BMAC (n = 8), and Group 2 (G2)—xenogeneic bone block combined with BMAC and hyperbaric oxygenation (n = 8). Bone biopsies were harvested 6 months after grafting. Vital Mineralized Tissue (VMT), Non-vital Mineralized Tissue (NVMT), and Non-Mineralized Tissue (NMT) were measured. Computed tomography was also performed on three occasions T0 (preoperative), T4 (4 months postoperative), and T8 (8 months postoperative). The difference between T4 and T8 values with respect to T0 was used to determine the thickness level gain after 4 and 8 months, respectively. Results: The tomographic evaluation did not show significant differences between the groups either at 4 or at the 8 months postoperatively. Regarding the histomorphometric analysis, CG had the lowest percentages of VMT (36.58 ± 9.56%), whereas G1 and G2 had similar results (55.64 ± 2.83% and 55.30 ± 1.41%, respectively). Concerning NMT and NVMT levels, the opposite was observed, with CG levels of 51.21 ± 11.54% and 11.16 ± 2.37%, G1 of 39.76 ± 11.48% and 3.65 ± 0.87%, and G2 of 40.3 ± 11.48% and 4.10 ± 0.87%, respectively. Conclusions: The use of bone block xenograft associated with BMAC resulted in a significant increase of bone neoformation when compared to the xenograft alone, though hyperbaric oxygenation did not enhance the results.Odontologí

A European Database of Fusarium graminearum and F-culmorum Trichothecene Genotypes

Fusarium species, particularly Fusarium graminearum and F culmorum, are the main cause of trichothecene type B contamination in cereals. Data on the distribution of Fusarium trichothecene genotypes in cereals in Europe are scattered in time and space. Furthermore, a common core set of related variables (sampling method, host cultivar, previous crop, etc.) that would allow more effective analysis of factors influencing the spatial and temporal population distribution, is lacking. Consequently, based on the available data, it is difficult to identify factors influencing chemotype distribution and spread at the European level. Here we describe the results of a collaborative integrated work which aims (1) to characterize the trichothecene genotypes of strains from three Fusarium species, collected over the period 2000-2013 and (2) to enhance the standardization of epidemiological data collection. Information on host plant, country of origin, sampling location, year of sampling and previous crop of 1147 F graminearurn, 479 F culmorum, and 3 F cortaderiae strains obtained from 17 European countries was compiled and a map of trichothecene type B genotype distribution was plotted for each species. All information on the strains was collected in a freely accessible and updatable database (www.catalogueeu.luxmcc.lu), which will serve as a starting point for epidemiological analysis of potential spatial and temporal trichothecene genotype shifts in Europe. The analysis of the currently available European dataset showed that in F. grarninearum, the predominant genotype was 15-acetyldeoxynivalenol (15-ADON) (82.9%), followed by 3-acetyldeoxynivalenol (3-ADON) (13.6%), and nivalenol (NIV) (3.5%). In F culmorum, the prevalent genotype was 3-ADON (59.9%), while the NIV genotype accounted for the remaining 40.1%. Both, geographical and temporal patterns of trichothecene genotypes distribution were identified.Ministere de l'Agriculture, de la Viticulture et de la Protection des Consommateurs-Administration des Services Techniques de l'Agriculture; M.I.U.R. Project AGROGEN (Laboratory of GENomics for traits of AGROnomic importance in durum wheat: Identification of useful genes, functional analysis and assisted selection by biological markers for the development of the national seed chain) [602/Ric]; Felix Thornley Cobbold Trust; John Oldacre Foundation; Ministry of Agriculture of the Czech RepublicMinistry of Agriculture, Czech Republic [800415]; Spanish Ministry MINECOSpanish Government [AGL201.4-53928-C2-2-R]; Ministry of Agriculture and Food, Norway; Federal Ministry of Education and Research (BMBF) (GABI-KANADA), BonnFederal Ministry of Education & Research (BMBF) [FKZ 0313711A]; German Academic Exchange Service (DAAD), BonnDeutscher Akademischer Austausch Dienst (DAAD) [A/06/92183]; Finnish Ministry of Agriculture and Forestry; Direction Generale de l'Agriculture, Direction de la Recherche [D31-3159, D31-1162, D31-7055]; P.O.R. SARDEGNA F.S.; Danish Directorate for Food, Fisheries and Agri Business [FFS05-3]; Academy of FinlandAcademy of Finland [126917, 131957, 250904, 252162, 267188, 266984]; Olvi Foundation; Turku University Foundation; CIMO travel grant; Nordic network project New Emerging Mycotoxins and Secondary Metabolites in Toxigenic Fungi of Northern Europe - Nordic Research Board [090014]The Luxembourg institute of Science and Technology, LU, acknowledges the Ministere de l'Agriculture, de la Viticulture et de la Protection des Consommateurs-Administration des Services Techniques de l'Agriculture for financially supporting the Sentinelle project. The work on Italian strains has been financially supported through the M.I.U.R. Project AGROGEN (Laboratory of GENomics for traits of AGROnomic importance in durum wheat: Identification of useful genes, functional analysis and assisted selection by biological markers for the development of the national seed chain) (D. D. 14.03.2005 n. 602/Ric). Funding for the research of Ryan Basler was provided by Felix Thornley Cobbold Trust and the John Oldacre Foundation.; The work of JC was supported by the Ministry of Agriculture of the Czech Republic, Project No. 800415. The research of MG and PG was supported by the Spanish Ministry MINECO (AGL201.4-53928-C2-2-R). The Ministry of Agriculture and Food, Norway funded the work of IH. The research of TM was funded by the Federal Ministry of Education and Research (BMBF) (GABI-KANADA #FKZ 0313711A), Bonn and by the German Academic Exchange Service (DAAD), Bonn (code no.: A/06/92183). PP acknowledges the Finnish Ministry of Agriculture and Forestry for funding the project FinMyco on Fusarium and mycotoxins in Finland. The research of JS was funded by the Direction Generale de l'Agriculture, Direction de la Recherche (ref. D31-3159, D31-1162, D31-7055), in the framework of a project entitled Caracterization et dynamique des fusarioses sur mais en Region Wallonne. BS acknowledges support by P.O.R. SARDEGNA F.S.E. 2007-2013-Obiettivo competitivita regionale e occupazione, Asse IV Capitale umano, Linea di Attivita 1.3.1 (research project Identification of natural and natural-like molecules inhibiting mycotoxin biosynthesis by Fusaria pathogenic on cereals). UT thanks the Danish Directorate for Food, Fisheries and Agri Business grant FFS05-3 for financial support. The work of TY was financially supported by the Academy of Finland (no. 126917, 131957, 250904, 252162, 267188, and 266984), Olvi Foundation, Turku University Foundation, a CIMO travel grant to Taha Hussien, and the Nordic network project New Emerging Mycotoxins and Secondary Metabolites in Toxigenic Fungi of Northern Europe (project 090014), which was funded by the Nordic Research Board