Generation of FGF reporter transgenic zebrafish and their utility in chemical screens

<p>Abstract</p> <p>Background</p> <p>Fibroblast Growth Factors (FGFs) represent a large family of secreted proteins that are required for proper development and physiological processes. Mutations in mouse and zebrafish FGFs result in abnormal embryogenesis and lethality. A key to understanding the precise role for these factors is to determine their spatial and temporal activity during embryogenesis.</p> <p>Results</p> <p>Expression of <it>Dual Specificity Phosphatase 6 </it>(<it>dusp6</it>, also known as <it>Mkp3</it>) is controlled by FGF signalling throughout development. The <it>Dusp6 </it>promoter was isolated from zebrafish and used to drive expression of destabilized green fluorescent protein (<it>d2EGFP</it>) in transgenic embryos (<it>Tg(Dusp6:d2EGFP)</it>). Expression of d2EGFP is initiated as early as 4 hours post-fertilization (hpf) within the future dorsal region of the embryo, where <it>fgf3 </it>and <it>fgf8 </it>are initially expressed. At later stages, d2EGFP is detected within structures that correlate with the expression of <it>Fgf </it>ligands and their receptors. This includes the mid-hindbrain boundary (MHB), pharyngeal endoderm, otic vesicle, hindbrain, and Kupffer's vesicle. The expression of d2EGFP is under the control of FGF signalling as treatment with FGF Receptor (FGFR) inhibitors results in the suppression of d2EGFP expression. In a pilot screen of commercially available small molecules we have evaluated the effectiveness of the transgenic lines to identify specific FGF inhibitors within the class of indolinones. These compounds were counter screened with the transgenic line <it>Tg(Fli1:EGFP)</it>^<it>y</it>1, that serves as an indirect read-out for Vascular Endothelial Growth Factor (VEGF) signalling in order to determine the specificity between related receptor tyrosine kinases (RTKs). From these assays it is possible to determine the specificity of these indolinones towards specific RTK signalling pathways. This has enabled the identification of compounds that can block specifically the VEGFR or the FGFR signalling pathway.</p> <p>Conclusion</p> <p>The generation of transgenic reporter zebrafish lines has allowed direct visualization of FGF signalling within the developing embryo. These FGF reporter transgenic lines provide a tool to screen for specific compounds that can distinguish between two conserved members of the RTK family.</p

RELATIONSHIP BETWEEN UPPER EXTREMITY MASS DISTRIBUTION AND PERFORMANCE IN COLLEGIATE TENNIS PLAYERS

The purpose of this study was to assess the relationship between upper extremity mass distribution and performance in tennis. Eighteen healthy collegiate tennis players consented to participate in this study. The study consisted of body mass distribution analysis using a dual X-ray absorptiometry scan and a simulated performance assessment to gauge accuracy and velocity of various tennis shots. The data collected suggests that mass distribution in the upper extremity is related to both shot velocity and accuracy

Parkinsonian motor impairment predicts personality domains related to genetic risk and treatment outcomes in schizophrenia

Identifying endophenotypes of schizophrenia is of critical importance and has profound implications on clinical practice. Here we propose an innovative approach to clarify the mechanims through which temperament and character deviance relates to risk for schizophrenia and predict long-term treatment outcomes. We recruited 61 antipsychotic naïve subjects with chronic schizophrenia, 99 unaffected relatives, and 68 healthy controls from rural communities in the Central Andes. Diagnosis was ascertained with the Schedules of Clinical Assessment in Neuropsychiatry; parkinsonian motor impairment was measured with the Unified Parkinson’s Disease Rating Scale; mesencephalic parenchyma was evaluated with transcranial ultrasound; and personality traits were assessed using the Temperament and Character Inventory. Ten-year outcome data was available for ~40% of the index cases. Patients with schizophrenia had higher harm avoidance and self-transcendence (ST), and lower reward dependence (RD), cooperativeness (CO), and self-directedness (SD). Unaffected relatives had higher ST and lower CO and SD. Parkinsonism reliably predicted RD, CO, and SD after correcting for age and sex. The average duration of untreated psychosis (DUP) was over 5 years. Further, SD was anticorrelated with DUP and antipsychotic dosing at follow-up. Baseline DUP was related to antipsychotic dose-years. Further, ‘explosive/borderline’, ‘methodical/obsessive’, and ‘disorganized/schizotypal’ personality profiles were associated with increased risk of schizophrenia. Parkinsonism predicts core personality features and treatment outcomes in schizophrenia. Our study suggests that RD, CO, and SD are endophenotypes of the disease that may, in part, be mediated by dopaminergic function. Further, SD is an important determinant of treatment course and outcome

Resignificando el sentido de la extensión a través del programa universitario en gestión bancaria

La Facultad de Ciencias Económica ha desarrollado, desde mediados del año 2017 y durante el año 2018, una experiencia de capacitación virtual cuyos destinatarios fueron hasta el momento 1060 trabajadores del Banco de la Provincia de Córdoba.Este artículo tiene como objetivo describir la propuesta, mostrando el modelo de educación a distancia que generamosen consonancia con el significado que otorgamos a la extensión universitaria. Además, trata de sistematizar la experiencia dando cuenta de los sentidos que se fueron construyendo durante su implementación, es decir, los efectos inimaginables que fueron dando forma a los intercambios y al trabajo colectivo entre los múltiples actores involucrados.El Programa Universitario de Gestión Bancaria es mucho más que un curso virtual. Ha sido la posibilidad de poner en acto un paradigma extensionista que contribuye al diálogo de saberes, a la democratización del conocimiento académico y a la revalorización del saber construido desde la práctica. Intercambio de saberes que priorizó el encuentro entre personas, que abrió puertas de instituciones tradicionalmente distantes y que le otorgó un sentido político a las prácticas extensionistas en la Facultad de Ciencias Económicas

EVALUACIÓN IN VITRO DE LA REMOCIÓN DE PLOMO EN AGUAS RESIDUALES POR Photobacterium damselae

Con la finalidad de mitigar los impactos ambientales ocasionados por plomo en aguas residuales de sectores como: minero, petroquímico, metalúrgico, otros, se realizó una evaluación in vitro de la remoción de plomo utilizando Photobacterium damselae. En la fase de aislamiento y bioestimulación las cepas obtenidas fueron sometidas a un proceso de selección en un medio de cultivo modificado, al cual se añadieron concentraciones de 20 y 100 ppm de Pb, obteniendo finalmente la cepa pura que mostró resistencia y/o tolerancia al Pb. La determinación de la capacidad para remover Pb en aguas residuales se observó en dos condiciones: incubación en temperatura controlada (25 C) e incubación en temperatura ambiente de Quito-Ecuador (zona sur a 2 800 msnm). La caracterización bioquímica de la bacteria fue realizada utilizando el Kit de Microgen GN-ID A+B. En la elaboración de las curvas de cinética de crecimiento bacteriano y remoción de Pb, se utilizaron técnicas de turbidimetría y absorción atómica, se destaca que Photobacterium damselae presentó una mayor facilidad de crecimiento a un máximo de 72 horas y a una concentración de 20 ppm en incubación a temperatura ambiente lográndose una remoción de hasta el 69% del plomo en el medio. De lo cual se infiere el potencial que tiene esta bacteria y se abren oportunidades para continuar estudios a futuro.// In order to mitigate the environmental impacts caused by lead in wastewater from sectors such as: mining, petrochemical, metallurgical and others, an in vitro evaluation of lead removal using Photobacterium damselae was carried out. Considering isolation and biostimulation phase, obtained isolate was subjected to a selection process in a modified culture medium, to which concentrations of 20 and 100 ppm of Pb were added, finally obtaining the pure strain that showed resistance and /or tolerance to Pb. To determinate the remotionÂŽs capacity of Pb in wastewater two conditions were observed: incubation at controlled temperature (25 C) and incubation at room temperature in Quito- Ecuador (southern zone at 2800msnm). Biochemical characterization of the bacteria was performed using the GN-ID A + B Microgen Kit. In the development of bacterial growth kinetics curves and Pb removal curves, turbidimetry and atomic absorption techniques were used, it was noted that Photobacterium damselae presented a greater rate of growth to a maximum of 72 hours and a concentration of 20 ppm in incubation at room temperature achieving a removal rate of 69% of the lead in the medium. From this information, the potential of this bacterium is inferred and opportunities are opened to continue studies in the future

Validation and reconstruction of flow meter data in the Barcelona water distribution network

12 páginas, 16 figuras, 1 tabla.-- El PDF es la versión pre-print.-- et al.This paper presents a signal analysis methodology to validate (detect) and reconstruct the missing and false data of a large set of flow meters in the telecontrol system of a water distribution network. The proposed methodology is based on two time-scale forecasting models: a daily model based on a ARIMA time series, while the 10-min model is based on distributing the daily flow using a 10-min demand pattern. The demand patterns have been determined using two methods: correlation analysis and an unsupervised fuzzy logic classification, named LAMDA algorithm. Finally, the proposed methodology has been applied to the Barcelona water distribution network, providing very good results.This work is part of a applied research project granted by ADASA and AGBAR companies. The authors also wish to thank the support received by the Research Commission of the Generalitat of Catalunya (Group SAC Ref. 2009 SGR 1491) and by CICYT (Ref. HYFA DPI2008-01996 and WATMAN DPI2009-13744) of Spanish Ministry of Education.Peer reviewe

Validation and reconstruction of flow meter data in the Barcelona water distribution network

This paper presents a signal analysis methodology to validate (detect) and reconstruct the missing and false data of a large set of flow meters in the telecontrol system of a water distribution network. The proposed methodology is based on two time-scale forecasting models: a daily model based on a ARIMA time series, while the 10-min model is based on distributing the daily flow using a 10-min demand pattern. The demand patterns have been determined using two methods: correlation analysis and an unsupervised fuzzy logic classification, named LAMDA algorithm. Finally, the proposed methodology has been applied to the Barcelona water distribution network, providing very good results.Peer ReviewedPostprint (author’s final draft

Variability in chemical profile and bioactivities of the flesh of greek pumpkin landraces

The aim of this study was to evaluate the chemical profile of the flesh and yield parameters of Greek pumpkin genotypes, including nine local landraces and two commercially available cultivars, focusing on valorizing the genetic pool of Cucurbita sp. with high added value products. Yield parameters (mean fruit weight and total fruit yield) recorded high variability with genotypes V8 and V2 showing the highest fruit yield. Moreover, genotype V11 was the most abundant in glucose and total sugars and scored the highest sweetness index suggesting good taste and promising marketing attributes. The highest antioxidant activity (OxHLIA assay) was assessed in the V8 genotype, while the V2 genotype showed the highest -, - and total tocopherols content. Oxalic acid was the main organic acid, followed by malic and citric acids, while organic acid composition varied among the tested genotypes. Moreover, the flesh extracts showed varied antimicrobial activity against several bacteria and fungi, while no toxicity against non-tumor cells was recorded. In conclusion, our results make evident the presence of high innate variability in terms of crop performance, chemical composition and bioactive properties not only between the different genotypes but also at the intrapopulational level. This finding is of high importance for the valorization of the local genetic pool of Cucurbita species through the selection of elite genotypes with high yield and quality of fruit, contributing to the conservation of valuable genetic material and limitation of the risk of genetic erosion due to neglect of local landraces.This research was funded by the Foundation for Science and Technology (FCT), P.I. (FCT, Portugal) through national funds FCT/MCTES to CIMO (UIDB/00690/2020 and UIDP/00690/2020), SusTEC (LA/P/0007/2021) and UIDB/50006/2020. National funding by FCT, P.I., through the institutional scientific employment program contract for M.I.D., R.C.C., C.P. and L.B. contracts, and M.G. and A.K.M. PhD grants (2020.06706.BD and 2020.06231.BD, respectively); FCT, P.I., within the scope of the Project PRIMA Section 2—Multi-topic 2019: PulpIng (PRIMA/0007/2019); the MICINN for the Juan de la Cierva Formación contract for T.C.S.P.P. (FJC20120-045405-I); and the General Secretariat for Research and Technology of the Ministry of Development and Investments under the PRIMA Program. PRIMA is an Art.185 initiative supported and co-funded under Horizon 2020, the European Union’s Program for Research and Plants 2022, 11, 800 16 of 18 Innovation (Prima2019-08).info:eu-repo/semantics/publishedVersio

Mutational screening of the TPO and DUOX2 genes in Argentinian children with congenital hypothyroidism due to thyroid dyshormonogenesis

[Purpose]: Primary congenital hypothyroidism (CH) is the most common endocrine disease in children and one of the preventable causes of both cognitive and motor deficits. We present a genetic and bioinformatics investigation of rational clinical design in 17 Argentine patients suspected of CH due to thyroid dyshormonogenesis (TDH). [Methods]: Next-Generation Sequencing approach was used to identify variants in Thyroid Peroxidase (TPO) and Dual Oxidase 2 (DUOX2) genes. A custom panel targeting 7 genes associated with TDH [(TPO), Iodothyrosine Deiodinase I (IYD), Solute Carrier Family 26 Member 4 (SLC26A4), Thyroglobulin (TG), DUOX2, Dual Oxidase Maturation Factor 2 (DUOXA2), Solute Carrier Family 5 Member 5 (SLC5A5)] and 4 associated with thyroid dysembryogenesis [PAX8, FOXE1, NKX2-1, Thyroid Stimulating Hormone Receptor (TSHR)] has been designed. Additionally, bioinformatic analysis and structural modeling were carried out to predict the disease-causing potential variants. [Results]: Four novel variants have been identified, two in TPO: c.2749-2 A > C and c.2752_2753delAG, [p.Ser918Cysfs*62] and two variants in DUOX2 gene: c.425 C > G [p.Pro142Arg] and c.2695delC [p.Gln899Serfs*21]. Eighteen identified TPO, DUOX2 and IYD variants were previously described. We identified potentially pahogenic biallelic variants in TPO and DUOX2 in 7 and 2 patients, respectively. We also detected a potentially pathogenic monoallelic variant in TPO and DUOX2 in 7 and 1 patients respectively. [Conclusions]: 22 variants have been identified associated with TDH. All described novel mutations occur in domains important for protein structure and function, predicting the TDH phenotype.This study was funded by grants from the Fondo para la Investigación Científica y Tecnológica (FONCyT-ANPCyT-MINCyT, PICT 2014-1193 to CMR, PICT 2015-1811 and PICT-2018-02146 to H.M.T.), CONICET (PIP 2015-11220150100499 to C.M.R.), Universidad de Buenos Aires (UBACyT 2016-20020150100099BA and 2020-20020190100050BA to C.M.R.) and Fondo de Investigación Sanitaria/FEDER (PI16/01920 and PI20/01589 to R.G.-S.)

Natural Distribution of Parasitoids of Larvae of the Fall Armyworm, Spodoptera frugiperda, in Argentina

To develop a better understanding of the natural distribution of the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), and to update the knowledge of the incidence of its complex of parasitoids. S. frugiperda, samplings in whorl-stage corn were carried out in provinces of Argentina from 1999 to 2003. S. frugiperda larvae were collected from corn in localities of the provinces of Tucumán, Salta, Jujuy, Santiago del Estero, La Rioja, Córdoba, San Luis, Chaco and Misiones. In each locality 30 corn plants were sampled and only larvae located in those plants were collected. The parasitoids that emerged from S. frugiperda larvae were identified and counted. The abundance of the parasitoids and the parasitism rate were estimated. The S. frugiperda parasitoids collected were Campoletis grioti (Blanchard) (Hymenoptera: Ichneumonidae), Chelonus insularis (Cresson) (Hymenoptera: Braconidae), Archytas marmoratus (Townsend) (Diptera Tachinidae) and/or A. incertus (Macquart), Ophion sp. (Hymenoptera: Ichneumonidae), Euplectrus platyhypenae Howard (Hymenoptera: Eulophidae), and Incamyia chilensis (Aldrich) (Diptera Tachinidae). C. grioti was the most abundant and frequent during the five-year survey. Similar diversity of parasitoids was obtained in all the provinces, with the exception of I. chilensis and E. platyhypenae that were recovered only in the province of Salta. In the Northwestern region, in Tucumán, C. grioti and species of Archytas were the most abundant and frequent parasitoids. On the contrary, in Salta and Jujuy Ch. insularis was the parasitoid most abundant and frequently recovered. The parasitism rate obtained in Tucumán, Salta and Jujuy provinces were 21.96%, 17.87% and 6.63% respectively with an average of 18.93%. These results demonstrate that hymenopteran and dipteran parasitoids of S. frugiperda occurred differentially throughout the Argentinian provinces and played an important role on the natural control of the S. frugiperda larval population