Etude de l'activité antivirale d'extraits d'Euphorbia de Corse : recherche de nouveaux diterpènes d'intérêt biologique

Chikungunya fever is caused by an arthropod-borne virus that is associated with massive epidemics and severe morbidity (virus-induced arthralgia, fever, myalgia and rashes). Worldwide expansion of the mosquito vectors, such as Aedes albopictus ("Tiger moquito) is responsible for the spread of Chikungunya virus (CHIKV) throughout the world. A. albopictus has spread throughout Mediterranean areas, which could lead to epidemics outbreaks. Currently, no antiviral drugs or vaccines are available for the treatment or prevention of CHIKV infection. Since ten years, however, recent results showed that diterpene esters from Trignostemon (Euphorbiaceae) possess inhibiting activity of CHIKV replication.With the objective to discover new compounds with antiviral activities, 45 extracts from various plant parts of 11 Euphorbiaceae species native to Corsica were evaluated for selective inhibition of CHIKV replication. In collaboration with Dr. Leyssen (KU Leuven, Belgium), several extracts made from 10 Euphorbia species exhibited significant and selective anti-CHIKV activity in a virus-cell-based assay. The antiviral activities of 29 commercially available phorboïds were studied. Some phorboïds were potent inhibitors of CHIKV and human immunodeficiency virus (VIH) replication. Results allowed drawing new structure-activity relationships, which supported the hypothesis that PKC may be an important target in CHIKV replication. In order to confirm or infirm the presence of phorboïds with anti-CHIKV activity in Euphorbia extracts, a liquid chromatography (LC) coupled to linear ion trap mass spectrometry (MSn) method was developed using standard compounds. Application of this methodology indicated that none anti-CHIKV phorboïds was present in Euphorbia extracts. A second LC-MSn procedure was developed to profile untargeted phorboïdes. Results suggested that numerous other diterpene esters were present in the Euphorbia extracts. The species Euphorbia amygdaloides ssp. semiperfoliata was selected to perform a bioassay-guided purification procedure, which led to the isolation and identification of 14 jatrophane esters, including eight new components. Among them, antiviral evaluation indicated that one jatrophane ester was possessing anti-CHIKV and anti-HIV activities. Furthermore, the structure of an atypical jatrophane ester derivative, jatrohemiketal, was determined unambiguously through an original strategy combining NMR spectroscopy and molecular modeling. Finally, an original tandem mass spectrometry (MS/MS)-targeted supercritical fluid chromatography (SFC) method was developed and used to study bioactive fractions of E. amygdaloïdes ssp. semiperfoliata. The MS/MS data were analyzed by molecular networking. Thanks to this approach, four 4-deoxyphorbol esters and two new jatrophane esters were targeted, isolated and identified. Collaboration with the group of Pr. Alcami (CNM, Espagne) was started to explore the anti-HIV properties of the isolated diterpene esters. Thus, one compound derived from 4-deoxyphorbol esters proved to possess a potent inhibiting activity of HIV-1 replication (IC50 = 8 nM, and selectivity index > 6250). The investigation of the mechanism of this component indicated that it acted like prostratin, but with antiviral effect more than 28-fold. Furthermore, the evaluation of the anti-CHIKV activity indicated that another 4-deoxyphorbol derivative was one of the strongest inhibitor of CHIKV replication isolated up to date (EC50 = 0.34 ± 0.12 µM and selectivity index > 638).Le chikungunya est une maladie transmise par des moustiques du genre Aedes (dont A. albopictus, dit "moustique tigre"). Cette maladie provoque d'intenses fièvres et des douleurs articulaires chroniques fortement invalidantes. Les moustiques potentiellement vecteurs du virus du chikungunya (CHIKV) sont des espèces invasives qui, à la faveur du réchauffement climatique, se sont récemment implantés dans plusieurs régions du monde, dont la région méditerranéenne. Sa présence constitue un terreau favorable à la survenue d'épidémie. A l'heure actuelle, il n'existe ni vaccin, ni traitement médicamenteux efficace. Toutefois, des articles scientifiques ont récemment rapporté que des esters de diterpène isolés du genre Trigonostemon (Euphorbiaceae), avaient une activité inhibitrice de la réplication du CHIKV.Dans le cadre de ces travaux de thèse, des extraits de plantes du genre Euphorbia de Corse ont été étudiés dans le but d'isoler de nouvelles molécules douées d'activité antivirale sur la réplication du CHIKV. En collaboration avec le Dr. P. Leyssen (KU Leuven, Belgique), l'évaluation de l'activité anti-CHIKV de 45 extraits, obtenus à partir de 11 Euphorbiaceae de Corse, a permis de mettre en évidence la forte activité inhibitrice et sélective de des extraits d'espèces du genre Euphorbia in cellulo. L'activité antivirale d'une série de 27 diterpènes de type phorboïde, disponibles commercialement, a également été étudiée. Les résultats ont montré que certains dérivés avaient une forte activité inhibitrice de la réplication du CHIKV, mais aussi sur celle du virus de l'immunodéficience humaine (VIH). Ces études ont permis d'une part, de déduire des relations structure-activité inédites et d'autre part, de soutenir l'hypothèse d'un mécanisme d'action anti-CHIKV impliquant la modulation des protéines kinases C (PKCs) par les phorboïdes. Dans le but de confirmer ou d'infirmer la présence des phorboïdes dans les extraits d'Euphorbia, une première méthode utilisant la chromatographie liquide (LC) haute performance couplée à un spectromètre de masse à trappe d'ions (MSn), a été développée à partir des composés standards. L'application de cette méthodologie a révélé qu'aucun des phorboïdes ciblés n'était présent dans les extraits d'Euphorbia. Ainsi, une seconde procédure LC-MSn a été mise en œuvre afin de détecter - de manière non ciblée - différents types d’esters diterpéniques. L'utilisation de cette approche a révélé que de nombreux diterpènes, non-apparentés aux phorboïdes, étaient présents dans les extraits. Un extrait de l'espèce Euphorbia amygdaloides subsp. semiperfoliata a été sélectionné pour réaliser un fractionnement bio-guidé, aboutissant à l'isolement et l'identification de 14 esters de jatrophane, dont neuf nouveaux composés. Parmi eux, l'un s'est avéré inhiber la réplication du CHIKV et du VIH. Par ailleurs, la structure d'un ester de jatrophane atypique, le jatrohémicétal, a été élucidée grâce à une approche originale combinant modélisation moléculaire et spectroscopie par résonance magnétique nucléaire (RMN). Enfin, une nouvelle procédure de purification ciblée par spectrométrie de masse tandem (MS/MS) en chromatographie en phase fluide supercritique (SFC) a été développée et appliquée sur des fractions bioactives d'E. amygdaloïdes subsp. semiperfoliata. L’interprétation des données MS/MS s’est appuyée sur la génération de réseaux moléculaires. Par cette méthodologie, quatre nouvelles molécules ont pu être détectées, purifiées et identifiées ; il s’agit de deux nouveaux esters de jatrophane et de quatre esters dérivés du 4-déoxyphorbol. L'activité anti-VIH des constituants isolés a également pu être explorée dans le cadre d’une collaboration avec l'équipe du Pr. Alcami (CNM, Espagne). Ainsi, l’un des esters de 4-deoxyphorbol s’est révélé être doué d'un exceptionnel pouvoir inhibiteur de la réplication du VIH-1 (IC50 = 8 nM et index de sélectivité > 6250). Son mécanisme d'action semble s'apparenter à celui de la prostratine (molécule antivirale de référence) mais avec des propriétés antivirales environ 28 fois supérieur. Un deuxième ester de 4-déoxyphorbol s’est avéré être un des plus puissants inhibiteurs du CHIKV isolé à ce jour (EC50 = 0,34 ± 0,12 µM and SI > 638). Mots clés : Euphorbia, activité antivirale, chikungunya, diterpène, spectrométrie de mass

Performance and Carcass Characteristics of Feedlot Steers: Effects of Delayed Implanting and Programmed Feeding During the Growing Period

This experiment was conducted to determine the effect of programming the rate of gain and delaying the first implant in feedlot steers on feedlot performance and carcass characteristics. Ninety-six growing steers (269 ± 16.2 kg) were assigned to 12 pens in a completely randomized design. Treatments were implant (Synovex-S®; 20 mg estradiol benzoate and 200 mg progesterone; Fort Dodge Animal Health, Overland Park, KS) on d 1 or no implant and programmed feeding to gain at a slow (0.68 kg/d) or fast (1.14 kg/d) rate during the growing period; these treatments were randomly assigned (n = 8) to pens of steers in a 2 × 2 factorial arrangement. Steers were fed a growing diet and after 88 and 60 d (for steers fed to gain at a slow or fast rate, respectively), steers were transitioned to ad libitum consumption of a high concentrate finishing diet. Growing period implant treatments did not affect ADG but did affect (P\u3c0.01) gain efficiency during the finishing period. Feeding steers for a slow rate of BW gain during the growing period improved (P=0.062) gain efficiency in the finishing period (169 vs 145 g gain/kg feed). Correlation coefficients between fat thickness and marbling score obtained via ultrasound and fat thickness and marbling score measured at harvest were greater the closer the ultrasound measurements were made to the final harvest date. These data indicate that feeding level prior to the start of the finishing period may affect BW gain efficiency during the finishing period

Effect of Long-Term Nutrient Management Strategies for Pastures on Phosphorus in Surface Runoff and Soil Quality

Manure, whether mechanically applied or deposited by grazing animals, has been associated with increased risk of non-point source pollution, especially phosphorus. This is especially true in areas where the industry, especially poultry, has been concentrated in geographical areas that are grain deficient, resulting in a reliance on imported grain for poultry feed. Intensification has resulted in the production of large quantities of poultry manure, within relatively small geographical areas. Surplus litter is typically land applied as a nutrient source or used as an animal feed. The objective of this project was to evaluate the effects of long-term nutrient management strategies using poultry litter as a feed and fertiliser for grazed pasture systems in the Shenandoah Valley of Virginia on soil quality, selected soil chemical characteristics and P losses in surface runoff

Development of a tomato pomace biorefinery based on a CO2-supercritical extraction process for the production of a high value lycopene product, bioenergy and digestate

Tomato peels and seeds (TP) are the most abundant canning industry waste actually used to produce biogas. TP is rich in lycopene (lyc) and represent a more sustainable feedstock than tomato fruits actually employed. It was therefore chosen as feedstock together with supercritical CO2 extraction (SFE-CO2) technology to develop a TP-SFE-CO2 biorefinery, topic scarcely investigated. Two TP were tested and although TP-SFE-CO2 parameters were the same, lyc recoveries depended by peel structure changes occurred during pre -SFE-CO2 drying step. Higher moisture (102.7 g kg-1 wet weight) permitted 97 % lyc recovery and gave a water-in-oil emulsion as extract. Mass balance confirmed that lyc isomerisation did not cause lyc losses. After a significant oil extraction, exhaust TP showed a biodegradability 64% higher than the raw one, attributable to fibre structure disruption. The biorefinery proposed (SFE_CO2+anaerobic digestion) determined positive economic revenue (+787.9 \u20ac t-1 TP) on the contrary of the actual TP management

Dynamic optimization based on Fourier. Application to the biodiesel process

[EN] This work presents a novel methodology for the dynamic optimization of the biodiesel production process from vegetable oils in discontinuous mode. The proposed methodology has the particularity of using the Fourier series for the parameterization of the control action, and evolutionary algorithms for the optimization of parameters. The main advantages of this strategy are, on the one hand, that the profiles obtained are smooth, that is, continuous and differentiable, therefore they can be directly implemented in real systems, without the need to filter or soften the control signal; on the other hand, a minimum amount of parameters is required for optimization, avoiding over-parameterization, which can decrease the quality of the response. The proposed algorithms have been evaluated through simulations, obtaining very satisfactory results compared to those published in the literature.[ES] Este trabajo presenta una novedosa metodología para la optimización dinámica del proceso de producción de biodiesel a partir de aceites vegetales en modo discontinuo. La metodología propuesta tiene la particularidad de emplear la serie de Fourier para la parametrización de la acción de control, y algoritmos evolutivos para la optimización de parámetros. Las ventajas principales de esta estrategia son, por un lado, que los perfiles obtenidos son suaves, es decir, continuos y diferenciables, por lo tanto pueden implementarse directamente en sistemas reales, sin necesidad de filtrar o suavizar la señal de control; por otro lado, se requiere una mínima cantidad de parámetros para la optimización, evitando la sobre-parametrización, la cual puede disminuir la calidad de la respuesta. Los algoritmos propuestos han sido evaluados a través de simulaciones, obteniendo resultados muy satisfactorios comparados con los existentes en bibliografía.Agradecemos al Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET) por financiar este proyecto, y al Instituto de Ingeniería Química (IIQ) de la Universidad Nacional de San Juan (UNSJ) por su continua colaboración.Pantano, MN.; Fernández, MC.; Rodríguez, L.; Scaglia, GJ. (2020). Optimización dinámica basada en Fourier. Aplicación al proceso de biodiesel. Revista Iberoamericana de Automática e Informática industrial. 18(1):32-38. https://doi.org/10.4995/riai.2020.12920OJS3238181Benavides, P. T. & Diwekar, U., 2012a. Optimal control of biodiesel production in a batch reactor: Part I: Deterministic control. Fuel,94, 211- 217. https://doi.org/10.1016/j.fuel.2011.08.035Benavides, P. T. & Diwekar, U., 2012b. Optimal control of biodiesel production in a batch reactor: Part II: Stochastic control. Fuel,94, 218-226. https://doi.org/10.1016/j.fuel.2011.08.033Brásio, A. S., Romanenko, A., Leal, J., Santos, L. O. & Fernandes, N. C., 2013. Nonlinear model predictive control of biodiesel production via transesterification of used vegetable oils. Journal of Process Control,10,23, 1471-1479. https://doi.org/10.1016/j.jprocont.2013.09.023Cantrell, D. G., Gillie, L. J., Lee, A. F. & Wilson, K., 2005. Structure-reactivity correlations in MgAl hydrotalcite catalysts for biodiesel synthesis. Applied Catalysis A: General,2,287, 183-190. https://doi.org/10.1016/j.apcata.2005.03.027Fernández, Cecilia, M., Nadia Pantano, M., Rossomando, F. G., Alberto Ortiz, O. & Scaglia, G. J., 2019. State estimation and trajectory tracking control for a nonlinear and multivariable bioethanol production system. Brazilian Journal of Chemical Engineering,1,36, 421-437. https://doi.org/10.1590/0104-6632.20190361s20170379Fernández, C., Pantano, N., Godoy, S., Serrano, E. & Scaglia, G., 2019a. Optimización de Parámetros Utilizando los Métodos de Monte Carlo y Algoritmos Evolutivos. Aplicación a un Controlador de Seguimiento de Trayectoria en Sistemas no Lineales. Revista Iberoamericana de Automática e Informática industrial,1,16, 89-99. https://doi.org/10.4995/riai.2018.8796Fernández M. C., P. M. N., Rodriguez L., Scaglia G., 2020. State Estimation and Nonlinear Tracking Control Simulation Approach. Application to a Bioethanol Production System. Bioprocess and Biosystems Engineering,In press.Fernández, M. C., Pantano, M. N., Machado, R. A. F., Ortiz, O. A. & Scaglia, G. J., 2019b. Nonlinear multivariable tracking control: application to an ethanol process. International Journal of Automation and Control,4,13, 440-468. https://doi.org/10.1504/IJAAC.2019.10020240Fernández, M. C., Pantano, M. N., Rómoli, S., Patiño, H. D., Ortiz, O. A. & Scaglia, G. J., 2019c. An algebra approach for nonlinear multivariable fedbatch bioprocess control. International Journal of Industrial and Systems Engineering,1,33, 38-57. https://doi.org/10.1504/IJISE.2019.10023564Fernández, M. C., Pantano, M. N., Serrano, E. & Scaglia, G., 2020. Multivariable Tracking Control of a Bioethanol Process under Uncertainties. Mathematical Problems in Engineering,2020. https://doi.org/10.1155/2020/8263690Ho, Y., Mjalli, F. & Yeoh, H., 2010. Multivariable adaptive predictive model based control of a biodiesel transesterification reactor. Journal of Applied Sciences,12,10, 1019-1027. https://doi.org/10.3923/jas.2010.1019.1027Ignat, R. M. & Kiss, A. A., 2013. Optimal design, dynamics and control of a reactive DWC for biodiesel production. Chemical Engineering Research and Design,9,91, 1760-1767. https://doi.org/10.1016/j.cherd.2013.02.009Kreyszig, E. 1978. Introductory functional analysis with applications, Wiley New York.Mjalli, F. S., Kim San, L., Chai Yin, K. & Azlan Hussain, M., 2009. Dynamics and control of a biodiesel transesterification reactor. Chemical Engineering & Technology,1,32, 13-26. https://doi.org/10.1002/ceat.200800243Nagle, R. K., Saff, E. B. & Snider, A. D. 2001. Ecuaciones diferenciales y problemas con valores en la frontera, Pearson Educación.Nasir, N., Daud, W. R. W., Kamarudin, S. & Yaakob, Z., 2013. Process system engineering in biodiesel production: A review. Renewable and Sustainable Energy Reviews,22, 631-639. https://doi.org/10.1016/j.rser.2013.01.036Nearing, J., 2006. Mathematical tools for physics.Pantano, M. N., Fernández, M. C., Serrano, M. E., Ortiz, O. A. & Scaglia, G. J. E., 2018. Tracking Control of Optimal Profiles in a Nonlinear Fed-Batch Bioprocess under Parametric Uncertainty and Process Disturbances. Industrial & Engineering Chemistry Research,32,57, 11130-11140.https://doi.org/10.1021/acs.iecr.8b01791Pantano, M. N., Serrano, M. E., Fernández, M. C., Rossomando, F. G., Ortiz, O. A. & Scaglia, G. J., 2017. Multivariable Control for Tracking Optimal Profiles in a Nonlinear Fed-Batch Bioprocess Integrated with State Estimation. Industrial & Engineering Chemistry Research,20,56, 6043- 6056. https://doi.org/10.1021/acs.iecr.7b00831Rajarathinam, K., Gomm, J. B., Yu, D.-L. & Abdelhadi, A. S., 2016. PID controller tuning for a multivariable glass furnace process by genetic algorithm. International Journal of Automation and Computing,1,13, 64- 72. https://doi.org/10.1007/s11633-015-0910-1Salvi, B. & Panwar, N., 2012. Biodiesel resources and production technologies-A review. Renewable and Sustainable Energy Reviews,6,16, 3680-3689. https://doi.org/10.1016/j.rser.2012.03.050Santori, G., Di Nicola, G., Moglie, M. & Polonara, F., 2012. A review analyzing the industrial biodiesel production practice starting from vegetable oil refining. Applied energy,92, 109-132. https://doi.org/10.1016/j.apenergy.2011.10.031Tempo, R. & Ishii, H., 2007. Monte Carlo and Las Vegas Randomized Algorithms for Systems and Control: An Introduction. European Journal of Control,2-3,13, 189-203. https://doi.org/10.3166/ejc.13.189-203Wali, W., Al-Shamma'a, A., Hassan, K. H. & Cullen, J., 2012. Online geneticANFIS temperature control for advanced microwave biodiesel reactor. Journal of Process Control,7,22, 1256-1272. https://doi.org/10.1016/j.jprocont.2012.05.013Wali, W., Hassan, K., Cullen, J., Shaw, A. & Al-Shamma'a, A., 2013. Real time monitoring and intelligent control for novel advanced microwave biodiesel reactor. Measurement,1,46, 823-839. https://doi.org/10.1016/j.measurement.2012.10.004Zhang, M., Gao, Z., Zheng, T., Ma, Y., Wang, Q., Gao, M. & Sun, X., 2016. A bibliometric analysis of biodiesel research during 1991-2015. Journal of Material Cycles and Waste Management, 1-9. https://doi.org/10.1007/s10163-016-0575-zZhang, Y., Dube, M., McLean, D. & Kates, M., 2003. Biodiesel production from waste cooking oil: 1. 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Inhibition of StearoylCoA Desaturase Activity Blocks Cell Cycle Progression and Induces Programmed Cell Death in Lung Cancer Cells

Lung cancer is the most frequent form of cancer. The survival rate for patients with metastatic lung cancer is ∼5%, hence alternative therapeutic strategies to treat this disease are critically needed. Recent studies suggest that lipid biosynthetic pathways, particularly fatty acid synthesis and desaturation, are promising molecular targets for cancer therapy. We have previously reported that inhibition of stearoylCoA desaturase-1 (SCD1), the enzyme that produces monounsaturated fatty acids (MUFA), impairs lung cancer cell proliferation, survival and invasiveness, and dramatically reduces tumor formation in mice. In this report, we show that inhibition of SCD activity in human lung cancer cells with the small molecule SCD inhibitor CVT-11127 reduced lipid synthesis and impaired proliferation by blocking the progression of cell cycle through the G1/S boundary and by triggering programmed cell death. These alterations resulting from SCD blockade were fully reversed by either oleic (18:1n-9), palmitoleic acid (16:1n-7) or cis-vaccenic acid (18:1n-7) demonstrating that cis-MUFA are key molecules for cancer cell proliferation. Additionally, co-treatment of cells with CVT-11127 and CP-640186, a specific acetylCoA carboxylase (ACC) inhibitor, did not potentiate the growth inhibitory effect of these compounds, suggesting that inhibition of ACC or SCD1 affects a similar target critical for cell proliferation, likely MUFA, the common fatty acid product in the pathway. This hypothesis was further reinforced by the observation that exogenous oleic acid reverses the anti-growth effect of SCD and ACC inhibitors. Finally, exogenous oleic acid restored the globally decreased levels of cell lipids in cells undergoing a blockade of SCD activity, indicating that active lipid synthesis is required for the fatty acid-mediated restoration of proliferation in SCD1-inhibited cells. Altogether, these observations suggest that SCD1 controls cell cycle progression and apoptosis and, consequently, the overall rate of proliferation in cancer cells through MUFA-mediated activation of lipid synthesis

Proceso de enseñanza-entrenamiento de jóvenes en el fútbol: posibilidades a partir de un modelo ondulatorio

The design of a youth athlete development process in soccer must be coherent with the major context, considering challenges and potentialities, and qualifying tactical-technical competences. Considering that, this theoretical study aims to present a “wavy model” for soccer, adapted from Ibáñez (2011), as a supporter for the organization and systematization of the tactical-technical defensive and offensive game phases, as much as insure the coaching and assessment process.El proceso de enseñanza-entrenamiento del fútbol debe ser coherente con el contexto en que se realiza, considerando los problemas y potencialidades, en busca de calificar las acciones técnico-tácticas y proporcionar la mejora en el juego. En esta perspectiva, este estudio de naturaleza teórica tiene como objetivo presentar un modelo ondulatorio para la enseñanza-entrenamiento del fútbol. Así, se presenta el modelo ondulatorio para el fútbol, adaptado de Ibáñez (2011), como una manera de ayudar en la organización y sistematización de los contenidos táctico-técnicos de ataque y defensa durante las etapas de formación, así como orientar la aplicación y evaluación de este proceso

Neonatal presentation of ventricular tachycardia and a Reye-like syndrome episode associated with disturbed mitochondrial energy metabolism

BACKGROUND: Hyperammonemia, hypoglycemia, hepatopathy, and ventricular tachycardia are common presenting features of carnitine-acylcarnitine translocase deficiency (Mendelian Inheritance in Man database: *212138), a mitochondrial fatty acid oxidation disorder with a lethal prognosis. These features have not been identified as the presenting features of mitochondrial cytopathy in the neonatal period. CASE PRESENTATION: We describe an atypical presentation of mitochondrial cytopathy in a 2 day-old neonate. She presented with a Reye-like syndrome episode, premature ventricular contractions and ventricular tachycardia. Initial laboratory evaluation exhibited a large amount of 3-methylglutaconic acid on urine organic acid analysis, mild orotic aciduria and a nonspecific abnormal acylcarnitine profile. The evaluation for carnitine-acylcarnitine translocase deficiency and other fatty acid oxidation disorders was negative. The patient later developed a hypertrophic cardiomyopathy and continued to be affected by recurrent Reye-like syndrome episodes triggered by infections. A muscle biopsy exhibited signs of a mitochondrial cytopathy. During the course of her disease, her Reye-like syndrome episodes have subsided; however, cardiomyopathy has persisted along with fatigue and exercise intolerance. CONCLUSIONS: This case illustrates that, in the neonatal period, hyperammonemia and ventricular tachycardia may be the presenting features of a lethal carnitine-acylcarnitine translocase deficiency or of a mitochondrial cytopathy, associated with a milder clinical course. This association broadens the spectrum of presenting phenotypes observed in patients with disturbed mitochondrial energy metabolism. Also, the presence of 3-methylglutaconic aciduria suggests mitochondrial dysfunction and mild orotic aciduria could potentially be used as a marker of mitochondrial disease

Postnatal Development of Numbers and Mean Sizes of Pancreatic Islets and Beta-Cells in Healthy Mice and GIPRdn Transgenic Diabetic Mice

The aim of this study was to examine postnatal islet and beta-cell expansion in healthy female control mice and its disturbances in diabetic GIPRdn transgenic mice, which exhibit an early reduction of beta-cell mass. Pancreata of female control and GIPRdn transgenic mice, aged 10, 45, 90 and 180 days were examined, using state-of-the-art quantitative-stereological methods. Total islet and beta-cell volumes, as well as their absolute numbers increased significantly until 90 days in control mice, and remained stable thereafter. The mean islet volumes of controls also increased slightly but significantly between 10 and 45 days of age, and then remained stable until 180 days. The total volume of isolated beta-cells, an indicator of islet neogenesis, and the number of proliferating (BrdU-positive) islet cells were highest in 10-day-old controls and declined significantly between 10 and 45 days. In GIPRdn transgenic mice, the numbers of islets and beta-cells were significantly reduced from 10 days of age onwards vs. controls, and no postnatal expansion of total islet and beta-cell volumes occurred due to a reduction in islet neogenesis whereas early islet-cell proliferation and apoptosis were unchanged as compared to control mice. Insulin secretion in response to pharmacological doses of GIP was preserved in GIPRdn transgenic mice, and serum insulin to pancreatic insulin content in response to GLP-1 and arginine was significantly higher in GIPRdn transgenic mice vs. controls. We could show that the increase in islet number is mainly responsible for expansion of islet and beta-cell mass in healthy control mice. GIPRdn transgenic mice show a disturbed expansion of the endocrine pancreas, due to perturbed islet neogenesis

Genotype–phenotype correlations in individuals with pathogenic RERE variants

Heterozygous variants in the arginine-glutamic acid dipeptide repeats gene (RERE) have been shown to cause neurodevelopmental disorder with or without anomalies of the brain, eye, or heart (NEDBEH). Here, we report nine individuals with NEDBEH who carry partial deletions or deleterious sequence variants in RERE. These variants were found to be de novo in all cases in which parental samples were available. An analysis of data from individuals with NEDBEH suggests that point mutations affecting the Atrophin-1 domain of RERE are associated with an increased risk of structural eye defects, congenital heart defects, renal anomalies, and sensorineural hearing loss when compared with loss-of-function variants that are likely to lead to haploinsufficiency. A high percentage of RERE pathogenic variants affect a histidine-rich region in the Atrophin-1 domain. We have also identified a recurrent two-amino-acid duplication in this region that is associated with the development of a CHARGE syndrome-like phenotype. We conclude that mutations affecting RERE result in a spectrum of clinical phenotypes. Genotype–phenotype correlations exist and can be used to guide medical decision making. Consideration should also be given to screening for RERE variants in individuals who fulfill diagnostic criteria for CHARGE syndrome but do not carry pathogenic variants in CHD7