Niemann-Pick type C disease mutations of NPC1 gene and evidence of abnormal expression of some mutant alleles in fibroblasts

We analyzed Niemann-Pick type C disease 1 (NP44406) gene in 12 patients with Niemann-Pick type C disease by sequencing both cDNA obtained from fibroblasts and genomic DNA. All the patients were compound heterozygotes. We found 15 mutations, eight of which previously unreported. The comparison of cDNA and genomic DNA revealed discrepancies in some subjects. In two unrelated patients carrying the same mutations (P474L and nt 2972del2) only one mutant allele (P474L), was expressed in fibroblasts. The mRNA corresponding to the other allele was not detected even in cells incubated with cycloheximide. The promoter variants (−1026T/G and −1186T/C or −238 C/G), found to be in linkage with 2972del2 allele do not explain the lack of expression of this allele, as they were also found in control subjects. In another patient, (N1156S/Q922X) the N1156S allele was expressed in fibroblasts while the expression of the other allele was hardly detectable. In a fourth patient cDNA analysis revealed a point mutation in exon 20 (P1007A) and a 56 nt deletion in exon 22 leading to a frameshift and a premature stop codon. The first mutation was confirmed in genomic DNA; the second turned out to be a T→G transversion in exon 22, predicted to cause a missense mutation (V1141G). In fact, this transversion generates a donor splice site in exon 22, which causes an abnormal pre-mRNA splicing leading to a partial deletion of this exon. In some NPC patients, therefore, the comparison between cDNA and genomic DNA may reveal an unexpected expression of some mutant alleles of NPC1 gene

DESENVOLVIMENTO E VALIDAÇÃO DE UM DISPOSITIVO PARA MENSURAÇÃO DA FLEXIBILIDADE POR MEIO DO ACELERÔMETRO EM CRIANÇAS

The objectives of this study were to develop and validate a computerized device to measure joint flexibility; to measure flexibility in children in elementary school aged 8 to 10 years using the Wells bank; to compare the data obtained by the Wells bank and the computerized device developed in the present study and evaluate the usability of the device. 215 volunteers participated in this research, aged between 8 and 10 years, 63.72% (137) were female and 36.28% (78) were male, duly enrolled in an educational institution in the Alto Tietê region. For data collection, the volunteers performed the flexibility test using the Wells bank and the computerized device developed in this research to assess flexibility. The computerized device it was made on a 3D printing technology printer. A usability evaluation was carried out with 15 Physical Education teachers for the product developed through a usability questionnaire based on the System Usability Scale (SUS) form, which contains 10 multiple-choice questions. The data obtained by the objective measures determined were evaluated in terms of frequency, mean and standard deviation. The arithmetic means of the three measurements performed at each stage were calculated. Mann-Whitney and Kruskal-Wallis tests and Spearman's correlation were performed for the investigation. For all analyses, significant differences were considered p<0.05. It is concluded that the computerized device to measure joint flexibility is capable of evaluating the range of motion in degrees, and was qualified as easy to handle by professionals in the field of physical education.Los objetivos de este estudio fueron desarrollar y validar un dispositivo computarizado para medir la flexibilidad articular; medir la flexibilidad en niños en la escuela primaria de 8 a 10 años a través del banco Wells; comparar los datos obtenidos por la base de datos Wells y el dispositivo computarizado desarrollado en el presente estudio y evaluar la usabilidad del dispositivo. Un total de 215 voluntarios participaron en esta investigación, con edades de 8 a 10 años, 63,72% (137) mujeres y 36,28% (78) hombres, debidamente matriculados en una institución educativa de la región del Alto Tietê. Para la recolección de datos, los voluntarios realizaron la prueba de flexibilidad utilizando la base de datos Wells y el dispositivo computarizado desarrollado en esta investigación para evaluar la flexibilidad. El dispositivo fue hecho de una impresora de tecnología de impresión 3D. Se realizó una evaluación de usabilidad con 15 profesores de Educación Física para el producto desarrollado a través del cuestionario de usabilidad basado en el formulario System Usability Scale (SUS) que contiene 10 preguntas de opción múltiple. Los datos obtenidos por las mediciones objetivas determinadas fueron evaluados para la frecuencia, media y desviación estándar. Se calcularon las medias aritméticas de las tres mediciones realizadas en cada etapa. Las pruebas de Mann-Whitney y Kruskal-Wallis y la correlación de Spearman se realizaron para la investigación. Para todos los análisis, se consideraron diferencias significativas p<0,05. Se concluye que el dispositivo computarizado para medir la flexibilidad articular es capaz de evaluar el rango de movimiento en grados, y fue calificado fácil de manejar por profesionales de la educación física.Os objetivos deste estudo foram desenvolver e validar um dispositivo computadorizado para mensurar a flexibilidade articular; mensurar a flexibilidade em crianças no ensino fundamental de 8 a 10 anos por meio do banco de Wells; comparar os dados alcançados pelo banco de Wells e o dispositivo computadorizado desenvolvido no presente estudo e avaliar a usabilidade do dispositivo. Participaram desta pesquisa 215 voluntários, com idade de 8 a 10 anos, sendo 63,72% (137) do sexo feminino e 36,28% (78) do sexo masculino, devidamente matriculados em uma instituição de ensino na região do Alto Tietê. Para coleta de dados, os voluntários realizaram o teste de flexibilidade utilizando o banco de Wells e o dispositivo computadorizado desenvolvido nessa pesquisa para avaliar a flexibilidade.O dispositivo foi confeccionado numa impressora de tecnologia de impressão 3D. Foi realizada uma avaliação da usabilidade com 15 professores de Educação Física para o produto desenvolvido através da interação de usabilidade baseada no formulário System Sustainability Scale (SUS) que contém 10 questões de múltipla-escolha. Os dados obtidos por medidas objetivas determinadas foram avaliadas quanto à frequência, média e desvio padrão. Foram seguidas como médias aritméticas das três medidas realizadas em cada etapa. Para investigar realizou-se os testes de Mann-Whitney e de Kruskal-Wallis e a tradução de Spearman. Para todas as análises foram consideradas diferenças significativas p<0,05.Conclui-se que o dispositivo computadorizado para medir a flexibilidade articular é capaz de avaliar em graus a amplitude de movimento

Budding yeast ATM/ATR control meiotic double-strand break (DSB) levels by down-regulating Rec114, an essential component of the DSB-machinery

An essential feature of meiosis is Spo11 catalysis of programmed DNA double strand breaks (DSBs). Evidence suggests that the number of DSBs generated per meiosis is genetically determined and that this ability to maintain a pre-determined DSB level, or "DSB homeostasis", might be a property of the meiotic program. Here, we present direct evidence that Rec114, an evolutionarily conserved essential component of the meiotic DSB-machinery, interacts with DSB hotspot DNA, and that Tel1 and Mec1, the budding yeast ATM and ATR, respectively, down-regulate Rec114 upon meiotic DSB formation through phosphorylation. Mimicking constitutive phosphorylation reduces the interaction between Rec114 and DSB hotspot DNA, resulting in a reduction and/or delay in DSB formation. Conversely, a non-phosphorylatable rec114 allele confers a genome-wide increase in both DSB levels and in the interaction between Rec114 and the DSB hotspot DNA. These observations strongly suggest that Tel1 and/or Mec1 phosphorylation of Rec114 following Spo11 catalysis down-regulates DSB formation by limiting the interaction between Rec114 and DSB hotspots. We also present evidence that Ndt80, a meiosis specific transcription factor, contributes to Rec114 degradation, consistent with its requirement for complete cessation of DSB formation. Loss of Rec114 foci from chromatin is associated with homolog synapsis but independent of Ndt80 or Tel1/Mec1 phosphorylation. Taken together, we present evidence for three independent ways of regulating Rec114 activity, which likely contribute to meiotic DSBs-homeostasis in maintaining genetically determined levels of breaks

First report in italy of the exotic mosquito species Aedes (Finlaya) koreicus, a potential vector of arboviruses and filariae

BACKGROUND: In the Veneto region (north-eastern Italy) an entomological surveillance system has been implemented since the introduction of the Asian tiger mosquito (Aedes albopictus) in 1991. During the routine monitoring activity in a tiger mosquito-free area, an unexpected mosquito was noticed, which clearly did not belong to the recorded Italian fauna. FINDINGS: At the end of May 2011, twelve larvae and pupae were collected in a small village in Belluno province (Veneto region) from a single manhole. Ten adults reared in the laboratory were morphologically and genetically identified as Aedes (Finlaya) koreicus (Edwards, 1917), a species native to Southeast Asia. The subsequent investigations carried out in the following months in the same village provided evidence that this species had become established locally. Entomological and epidemiological investigations are currently ongoing in the surrounding area, to verify the eventual extension of the species outside the village and to trace back the route of entry into Italy. CONCLUSIONS: This is the first report in Italy of the introduction of the exotic mosquito Ae. koreicus. This species has been shown experimentally to be competent in the transmission of the Japanese encephalitis virus and of the dog heartworm Dirofilaria immitis and is considered a potential vector of other arboviruses. Thus, the establishment of this species may increase the current risk or pose new potential threats, for human and animal health. This finding considerably complicates the entomological monitoring of the Asian tiger mosquito Ae. albopictus in Italy and stresses the importance of implementing the entomological surveillance for the early detection of and the rapid response against invasive mosquito species

Exchange of nutrients and oxygen across the sediment-water interface below a Sparus aurata marine fish farm in the north-western Mediterranean Sea

Purpose: This study analyzes the effects of aquaculture activities in open seawater in the north-western coastal waters of the Mediterranean Sea. It is the first of its kind to be based on benthic flux data gathered in situ below fish farms for this particular area. Materials and methods: Samples were collected on four sampling campaigns over a 1-year cycle under a Sparus aurata fish farm facility where benthic fluxes were measured in situ using light and dark benthic chambers. Bottom water and sediment samples were also collected. Data were compared to those for a nearby control station. Results and discussion: Significant differences were found (ANOVA, p < 0. 05) between concentrations of organic matter (OM), total phosphorus and redox potentials in sediments located under the cages and those of the control station. The consumption of dissolved oxygen (DO) by sediment and positive ammonium (NH4 +) fluxes was stimulated by OM content, with correlations of r = -0. 60 (p < 0. 01) and r = 0. 70 (p < 0. 01), respectively. The OM content of sediments was found to be consistently higher under the cages than at the control station, with the highest value (1. 8 ± 0. 7 %) under the cages observed during the early summer; values of DO and NH4 + fluxes were -64 ± 17 and 12. 7 ± 1. 0 mmol m-2 day-1, respectively. PO4 3- fluxes were consistently higher in the fish farm sediments (between 0. 58 and 0. 98 mmol m-2 day-1) than those observed at the control station. Nitrate (NO3 -) fluxes were found to be consistently negative due to denitrification occurring in the sediments and were related to the concentration of NO3 - in bottom waters (r = 0. 92, p < 0. 01). Si fluxes were shown to be associated with water temperature (r = 0. 59, p < 0. 05). Conclusions: The results imply that sediments located below cages accumulate organic matter originating from aquaculture activities, especially during summer months when this activity increases. Sediments undergo biogeochemical changes that mainly affect fluxes of DO, NH4 + and soluble reactive phosphorus, although these do not seem to have a significant impact on the quality of the water column due to the hydrodynamic characteristics of the area. © 2012 Springer-Verlag.We would like to thank the Caja del Mediterraneo for a predoctoral fellowship fund for this research and Antonio Asuncion Acuigroup Maremar manager for the facilities and support in conducting the study. The translation of this paper was funded by the Universidad Politecnica de Valencia, Spain. We are grateful for the valuable comments of the anonymous reviewers on previous versions of the manuscript.Morata Higón, T.; Sospedra, J.; Falco Giaccaglia, SL.; Rodilla Alama, M. (2012). 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Serum IgG against Simian Virus 40 antigens are hampered by high levels of sHLA-G in patients affected by inflammatory neurological diseases, as multiple sclerosis

Background: Many investigators detected the simian polyomavirus SV40 footprints in human brain tumors and neurologic diseases and recently it has been indicated that SV40 seems to be associated with multiple sclerosis (MS) disease. Interestingly, SV40 interacts with human leukocyte antigen (HLA) class I molecules for cell entry. HLA class I antigens, in particular non-classical HLA-G molecules, characterized by an immune-regulatory function, are involved in MS disease, and the levels of these molecules are modified according with the disease status. Objective: We investigated in serum samples, from Italian patients affected by MS, other inflammatory diseases (OIND), non-inflammatory neurological diseases (NIND) and healthy subjects (HS), SV40-antibody and soluble sHLA-G and the association between SV40-prevalence and sHLA-G levels. Methods: ELISA tests were used for SV40-antibodies detection and sHLA-G quantitation in serum samples. Results: The presence of SV40 antibodies was observed in 6 % of patients affected by MS (N = 4/63), 10 % of OIND (N = 8/77) and 15 % of NIND (N = 9/59), which is suggestive of a lower prevalence in respect to HS (22 %, N = 18/83). MS patients are characterized by higher sHLA-G serum levels (13.9 \ub1 0.9 ng/ml; mean \ub1 St. Error) in comparison with OIND (6.7 \ub1 0.8 ng/ml), NIND (2.9 \ub1 0.4 ng/ml) and HS (2.6 \ub1 0.7 ng/ml) subjects. Interestingly, we observed an inverse correlation between SV40 antibody prevalence and sHLA-G serum levels in MS patients. Conclusion: The data obtained showed a low prevalence of SV40 antibodies in MS patients. These results seems to be due to a generalized status of inability to counteract SV40 infection via antibody production. In particular, we hypothesize that SV40 immune-inhibitory direct effect and the presence of high levels of the immune-inhibitory HLA-G molecules could co-operate in impairing B lymphocyte activation towards SV40 specific peptides