Role of Beta2 adrenergic receptor polymorphism (rs1042714) on body weight and glucose metabolism response to a meal-replacement hypocaloric diet

Producción CientíficaThe Beta2-adrenergic receptor (ADRB2) is involved in energy balance regulation. The objective of our study was to evaluate the role of rs1042714 genetic variant of ADRB2 gene on weight loss, body composition and metabolic changes secondary to partial Meal replacement (pMR) hypocaloric diet in women with obesity. G allele of rs rs1042714 predicts the magnitude of weight loss resulting from a pMR diet. These adiposity improvements produce a better improvement of insulin resistance and percentage of impaired glucose metabolism in G allele carriers

Role of the rs10401670 variant in the resistin gene on the metabolic response after weight loss secondary to a high‐fat hypocaloric diet with a Mediterranean pattern

Producción CientíficaBackground:The single nucleotide polymorphism (SNP) (rs10401670) of theRETNgene has been associated with metabolic disorder in obese subjects andhas scarcely been evaluated after dietary interventions. The present studyaimed to analyse the effects of thers10401670 RETNgene polymorphismon metabolic changes secondary to weight loss and secondary to a high‐fathypocaloric diet with a Mediterranean dietary pattern.Methods:A Caucasian population comprising 284 obese patients withoutdiabetes mellitus was analysed. Before and after 3 months of a high‐fathypocaloric diet with a Mediterranean pattern, an anthropometric evaluation,an assessment of nutritional intake and a biochemical analysis were per-formed. A statistical analysis was conducted for the combinedCTandTTas agroup and for wild‐typeCCas a second group.Results:Decreases in weight, body mass index (BMI), fat mass, systolic bloodpressure and waist circumference were similar in both genotypes groups. InTallele carriers, insulin, homeostatic model assessment for insulin resistance(HOMA‐IR), triglycerides and C‐reactive protein levels were decreased.The decrease in these parameters was statistically significant for triglycerides(−22.3 ± 9.3 mg dl–1:p= 0.03), C‐reactive protein (−2.8 ± 0.5 mg dl–1:p= 0.03), insulin (−7.4 ± 2.9 mUI L–1:p= 0.03) and HOMA‐IR (−2.4 ± 1.0:p= 0.02). Leptin levels were decreased in both genotypes groups after thehypocaloric diet, as well as the anthropometric parameters BMI, weight, waistcircumference and fat mass. Resistin and adiponectin levels remained un-changed in both groups.Conclusions:In the present study, we have detected a significant associationbetween theTallele of this SNP and a better response of insulin resistance,triglycerides and C‐reactive protein compared to nonTallele carriers afterweight loss with a high‐fat hypocaloric diet and a Mediterranean diet

Adiponectin gene variant rs3774261, effects on lipid profile and adiponectin levels after a high polyunsaturated fat hypocaloric diet with Mediterranean pattern

Producción CientíficaThe role of ADIPOQ gene variants on metabolic improvements after weight change secondary to different hypocaloric diets remained unclear. We evaluate the effect of rs3774261 of ADIPOQ gene polymorphism on biochemical improvements and weight change after high polyunsaturated fat hypocaloric diet with a Mediterranean dietary pattern for 12 weeks. A population of 361 obese subjects was enrolled in an intervention trial with a calorie restriction of 500 calories over the usual intake and 45.7% of carbohydrates, 34.4% of fats, and 19.9% of proteins. The percentages of different fats was; 21.8% of monounsaturated fats, 55.5% of saturated fats, and 22.7% of polyunsaturated fats. Before and after intervention, an anthropometric study, an evaluation of nutritional intake and a biochemical evaluation were realized. All patients lost weight regardless of genotype and diet used. After 12 weeks with a similar improvement in weight loss (AA vs. AG vs. GG); total cholesterol (delta: −28.1 ± 2.1 mg/dL vs. −14.2 ± 4.1 mg/dL vs. −11.0 ± 3.9 mg/dL; p = 0.02), LDL cholesterol (delta: −17.1 ± 2.1 mg/dL vs. −6.1 ± 1.9 mg/dL vs. −6.0 ± 2.3 mg/dL; p = 0.01), triglyceride levels (delta: −35.0 ± 3.6 mg/dL vs. 10.1 ± 3.2 mg/dL vs. −9.7 ± 3.1 mg/dL; p = 0.02), C reactive protein (CRP) (delta: −2.3 ± 0.1 mg/dL vs. −0.2 ± 0.1 mg/dL vs. −0.2 ± 0.1 mg/dL; p = 0.02), serum adiponectin (delta: 11.6 ± 2.9 ng/dL vs. 2.1 ± 1.3 ng/dL vs. 3.3 ± 1.1 ng/dL; p = 0.02) and adiponectin/leptin ratio (delta: 1.5 ± 0.1 ng/dL vs. 0.3 ± 0.2 ng/dL vs. 0.4 ± 0.3 ng/dL; p = 0.03), improved only in AA group. AA genotype of ADIPOQ variant (rs3774261) is related with a significant increase in serum levels of adiponectin and ratio adiponectin/leptin and decrease on lipid profile and C-reactive protein (CRP)

Effect of Two Meal Replacement strategies on Cardiovascular Risk Parameters in Advanced Age Patients with Obesity and Osteoarthritis

Producción CientíficaBackground and aims: Meal replacement diets consist of replacing one or more meals with an artificial nutritional supplement. The objective of this study was to compare the effect of one against two meal replacement strategies on body composition and cardiovascular risk parameters in patients with obesity. Methods: A randomized clinical trial was designed with a modified hypocaloric diet with an artificial nutritional preparation replacing one or two meals for three months in patients with obesity and osteoarthritis pending orthopedic surgery. An anthropometric evaluation and a measurement of the body composition were done with bioelectrical impedance measurement at the beginning and at three months. Results: A total of 112 patients were recruited. Fifty-two patients (46.4%) were randomized to one replacement and 60 patients (53.6%) to two meal replacements. Eighty-one patients (72.3%) were women, and the average age was 61 (11.03) years. The percentage of weight loss at three months was 8.27 (4.79)% (one meal replacement: 7.98 (5.97)%; two meal replacements: 8.50 (3.48)%; p = 0.56). A decrease in fat mass measured by the fat mass index (FMI) was detected (one meal replacement: −2.15 (1.45) kg/m2 vs. two meal replacements: −2.78 (2.55) kg/m2; p > 0.05), and a relative increase in fat-free mass was observed (one meal replacement: +3.57 (4.61)% vs. two meal replacements: +2.14 (4.45)%; p > 0.05). A decrease in HOMA-IR, systolic blood pressure (SBP), and total cholesterol was observed in both groups without differences between them. Conclusions: The substitution strategies of one or two meal replacements were effective in weight loss and fat mass decrease without differences between the two groups. An improvement in lipid parameters, glycemic control, and systolic blood pressure was observed without differences between strategies

Effects of a snack enriched with carob and Undaria pinnatifida (wakame) on metabolic parameters in a double blind, randomized clinical trial in obese patients

Introduction: Introduction: the composition of snack foods likely influences the overall effect that snacking has on metabolism and obesity. The objective of the current study was to assess the responses to two different snacks, one of them supplemented with wakame and carobs, on cardiovascular risk factors, satiety, and subsequent food intake in obese subjects with metabolic syndrome. Material and Methods: forty patients were randomized in a clinical trial (NCT03420989, clinicaltrial.gov) to group I (enriched snack, n = 16) or group II (control snack, n = 16). At baseline and after 8 weeks biochemical parameters, dietary intakes, and nutritional status were assessed. The subjects also rated their feelings of satiety/hunger with a test meal. Results: no differences were detected in anthropometric parameters between both snacks. Changes in other parameters were detected in patients with enriched snacks, with a significant decrease in LDL-cholesterol by 7.4 % (intervention snack, -8.9 ± 2.3 mg/dL vs control snack, -0.9 ± 3.3 mg/dL; p = 0.03), in total cholesterol by 5.8 % (intervention snack, -10.4 ± 2.9 mg/dL vs control snack, -1.4 ± 3.2 mg/dL; p = 0.02), and in resistin level by 15.9 % (intervention snack, -1.0 ± 0.2 mg/dL vs control snack, -0.1 ± 0.3 mg/dL: p = 0.03). After the test meal, satiety scores (after 20 min and 40 min) were higher than fasting levels in both groups. The same results were obtained with the 100-mm, 5-point visual satiety scale. Conclusion: our study indicates that a wakame- and carob-enriched snack induces a significant decrease in total cholesterol, LDL-cholesterol, and resistin levels when compared to a control snack, without effects on food consumption, other cardiovascular parameters, or anthropometric parameters.Introduccion: la composicion de los snacks probablemente influya en el efecto que produce su consumo sobre los marcadores metabolicos y la obesidad. El objetivo fue evaluar respuestas a dos snacks, uno de ellos suplementado con wakame y algarroba, sobre factores de riesgo cardiovascular, saciedad y posterior ingesta de alimentos, en sujetos obesos con sindrome metabolico. Material y metodos: se aleatorizaron 40 pacientes en el ensayo clinico NCT03420989 (clinicaltrial.gov) para participar en el grupo I (snack enriquecido, n = 16) o el grupo II (snack de control, n = 16). Antes y despues de 8 semanas se determinaron parametros bioquimicos, ingestas dieteticas y estado nutricional. A los sujetos tambien se les evaluo saciedad y apetito con comida de prueba. Resultados: no se detectaron diferencias en parametros antropometricos con ambos snacks. Se detectaron cambios en parametros bioquimicos de pacientes que recibieron snacks enriquecidos, con disminucion significativa del colesterol-LDL del 7,4 % (snack de intervencion, -8,9 ± 2,3 mg/dl vs. snack de control, -0,9 ± 3,3 mg/dl; p = 0,03), del colesterol total del 5,8 % (snack de intervencion, -10,4 ± 2,9 mg/dl vs. snack de control, -1,4 ± 3,2 mg/dl; p = 0,02) y de niveles de resistina del 15,9 % (snack de intervencion, -1,0 ± 0,2 mg/dl vs. snack de control, -0,1 ± 0,3 mg/dl; p = 0,03). Despues de la comida de prueba, las puntuaciones de saciedad (a los 20 min y 40 min) fueron mas altas que nivel de ayuno en ambos grupos. Los resultados fueron similares con escala de saciedad visual de 5 puntos y 100 mm. Conclusion: nuestro estudio muestra que un snack enriquecido con wakame y algarroba produce disminucion significativa de los niveles de colesterol total, colesterol-LDL y resistina frente a un snack de control, sin efectos sobre el consumo de alimentos, otros parametros cardiovasculares y los parametros antropometricos

Obesidad en España y Castilla y León: ¿qué nos dicen los estudios?

La obesidad, es la epidemia del siglo XXI, se define como un peso corporal desproporcionado para la altura con una acumulación excesiva de tejido adiposo que se acompaña habitualmente de una inflamación sistémica crónica leve. Para clasificar a los pacientes con obesidad se utiliza el índice de Quetelet o índice de masa corporal, que se obtiene dividiendo el peso del individuo en Kg entre la talla al cuadrado en metros (kg/m2). Un índice de masa corporal por encima de 30 kg/m2 se considera como obesidad. La obesidad está asociada con el desarrollo de diabetes mellitus tipo 2, enfermedades cardiovasculares, algunos tipos de cáncer y otras condiciones patológicas crónicas. La circunferencia de la cintura también es un marcador de riesgo, en obesidad. En el estudio ENPE, se demostró una prevalencia en la población española de sobrepeso de 39,3% y de obesidad de 21,6%. La prevalencia de sobrepeso en mujeres fue de 32,1% y de obesidad de 22,8%, en varones, por otra parte, la obesidad represento un 20,5% y el sobrepeso un 46,5 %. Recientemente, en Castilla y León, la Consejería de Sanidad ha publicado los resultados de un estudio de cohortes “Estudio de Riesgo Cardiovascular en Castilla y León; la evolución de los Factores de Riesgo cardiovascular 2004-2014”. La prevalencia de obesidad (IMC ≥30) en 2004 era en nuestra Comunidad Autónoma de 22,9%. En la reevaluación de la misma cohorte de pacientes, diez años más tarde, esa proporción ha aumentado hasta el 27,6%, casi 5 puntos porcentuales. Con respecto al total de la muestra de 2014, el 31,1% tenían normopeso, el 41,3% tenían sobrepeso y el 26,1% tenían un IMC entre 30 y 40 (obesidad) y un 1,6% presentaban un IMC>40.En conclusión, la obesidad se ha convertido en una pandemia a nivel mundial, situándose España en una zona intermedia de prevalencia, así como Castilla y León. EL índice de masa corporal, así como la circunferencia de la cintura son las herramientas más utilizadas a la hora de realizar los estudios de prevalencia de obesidad.Obesity, the epidemic of the 21st century (1), is defined as a disproportionate body weight for height with an excessive accumulation of adipose tissue that is usually accompanied by a mild chronic systemic inflammation. To classify patients with obesity, the Quetelet index or body mass index is used, which is obtained by dividing the weight of the individual in Kg by the square height in meters (kg/m2). A body mass index above 30 kg/m2 is considered as obesity. Obesity is associated with the development of type 2 diabetes mellitus, cardiovascular diseases, some types of cancer and other chronic pathological conditions. Waist circumference is a risk factor in obesity, too. In the ENPE study, a prevalence in the Spanish population of overweight of 39.3% and of obesity of 21.6% was demonstrated. The prevalence of overweight in women was 32.1% and obesity 22.8%, in men; on the other hand, obesity represented 20.5% and overweight 46.5%. Recently, Castilla y León Health System has published the results of a study of cohorts “Cardiovascular Risk Study in Castilla y León; the evolution of cardiovascular risk factors 2004-2014 “. The prevalence of obesity (BMI ≥30) in 2004 was 22.9% in our Autonomous Community. In the re-evaluation of the same cohort of patients, ten years later, that proportion has increased to 27.6%, almost 5 percentage points. With respect to the total sample of 2014, 31.1% had normal weight, 41.3% were overweight and 26.1% had a BMI between 30 and 40 (obesity) and a 1, 6% had a BMI> 40 In conclusion, obesity has become a pandemic worldwide, placing Spain in an intermediate zone of prevalence as well as Castilla y León. The body mass index, as well as the circumference of the waist are the most used tools when carrying out obesity prevalence studie

Influence of obesity on bone turnover markers and fracture risk in postmenopausal women

Producción CientíficaBackground and aims: The relationship between obesity and bone metabolism is controversial. In recent decades, the protective role of obesity in the development of osteoporosis is questioned. The aims of this study are the following: to evaluate the differences in bone turnover markers between postmenopausal women with and without obesity and to compare the risk of fracture at five years between these groups. Methods: An observational longitudinal prospective cohort study of postmenopausal women with obesity (O) (body mass index (BMI) > 30 kg/m2) and non-obesity (NoO) (BMI < 30 kg/m2) is designed. 250 postmenopausal women are included in the study (NoO: 124 (49.6%) and O: 126 (50.4%)). It measures epidemiological variables, dietary variables (calcium intake, vitamin D intake, smoking, alcohol consumption, and physical activity), biochemicals (β-crosslap, type I procollagen amino-terminal peptide (P1NP), 25OH-vitamin D, and parathyroid hormone (PTH)), anthropometric variables, and fracture data five years after the start of the study. The mean age is 56.17 (3.91) years. Women with obesity showed lower levels of vitamin D (O: 17.27 (7.85) ng/mL, NoO: 24.51 (9.60) ng/mL; p < 0.01), and higher levels of PTH (O: 53.24 (38.44–65.96) pg/mL, NoO: 35.24 (25.36–42.40) pg/mL; p < 0.01). Regarding the bone formation marker (P1NP), it was found to be high in women without obesity, O: 45.46 (34.39–55.16) ng/mL, NoO: 56.74 (45.34–70.74) ng/mL; p < 0.01; the bone resorption marker (β-crosslap) was found to be high in women with obesity, being significant in those older than 59 years (O: 0.39 (0.14) ng/mL, NoO 0.24 (0.09) ng/mL; p < 0.05). No differences are observed in the risk of fracture at 5 years based on BMI (OR = 0.90 (95%CI 0.30–2.72); p = 0.85). Conclusions: Postmenopausal women with obesity showed lower levels of bone formation markers; older women with obesity showed higher markers of bone resorption

Detection, molecular characterisation and aspects involving the transmission of tomato chlorotic dwarf viroid in eggplant

"This is the peer reviewed version of the following article: Gramazio P, Lerma MD, Villanueva G, et al. Detection, molecular characterisation and aspects involving the transmission of tomato chlorotic dwarf viroid in eggplant. Ann Appl Biol. 2019;175:172 183. https://doi.org/10.1111/aab.12527, which has been published in final form at https://doi.org/10.1111/aab.12527. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving."[EN] Tomato chlorotic dwarf viroid (TCDVd) is a pospiviroid that causes severe disease symptoms in tomato. TCDVd is also naturally found in other crops and plants, in most occasions being asymptomatic. Apart from the natural hosts reported up to now, artificial inoculations have revealed that TCDVd can infect other plants, including eggplant (Solanum melongena). In a screening of seedlings of eggplant from a breeding programme we detected a pospiviroid, which we identified as TCDVd, representing the first report of natural infection of eggplant by TCDVd. The new TCDVd isolate of eggplant was detected by reverse transcription polymerase chain reaction (RT¿PCR) using primers TG21/CT20, initially designed to detect potato spindle tuber viroid. The new isolate sequence is close to a Brugmansia sanguinea isolate of TCDVd from the Netherlands, and most of the nucleotidic changes with respect to this isolate and to the reference genome sequence of TCDVd are found in the TR region. Naturally infected plants of eggplant with this TCDVd isolate did not display any disease symptoms. We demonstrated that in eggplant TCDVd is mechanically transmitted with low to moderate efficiency with cultivation practices, but not by plant¿to¿plant contact. Tomato plants artificially inoculated with the eggplant isolate of TCDVd tested positive for the presence of the viroid at 50¿days after inoculation, but did not display any disease symptoms. Seed transmission to germinated seedlings of eggplant was variable among progenies from infected plants, ranging from 7.7% to 100.0%. Disinfection of seeds with chemical treatments with sodium hypochlorite and trisodium phosphate solutions plus thermotherapy at 80°C for 24¿hr or 90°C for 6 hr was ineffective in reducing the rate of transmission by seed. We did not find evidence of horizontal transmission of TCDVd by pollen, but vertical transmission was highly efficient when healthy eggplant plants were pollinated with infected pollen. Our results indicate that asymptomatic infection of eggplant by TCDVd and high seed and pollen transmission rates may contribute to the spread of this viroid. The information we obtained is useful in order to implement measures for the prevention, control and eradication of TCDVd in eggplant crops, as well as to avoid their transmission to other hosts.P.G. is grateful to Universitat Politècnica de València and to Japan Society for the Promotion of Science for their respective postdoctoral grants (PAID-10-18 and FY2019 JSPS Postdoctoral Fellowship for Research in Japan [Standard]). A.A. and D.A. are grateful to Universitat Politècnica de València for their respective predoctoral (PAID-01-18 and PAID-01-16) contracts within the Programa de Ayudas de Investigación y Desarrollo initiative. E.G.-F. is grateful to Ministerio de Educación, Cultura y Deporte para la Formación de Profesorado Universitario for a predoctoral grant (FPU17/02389). G.M. is grateful to Generalitat Valenciana for a predoctoral grant within the Santiago Grisolía programme (GRISOLIAP/2016/012). M.P. is grateful to Generalitat Valenciana and Fondo Social Europeo for a postdoctoral grant (APOSTD/2018/014).Gramazio, P.; Lerma Lerma, MD.; Villanueva-Párraga, G.; Vilanova Navarro, S.; García-Fortea, E.; Mangino, G.; Figás-Moreno, MDR.... (2019). Detection, molecular characterisation and aspects involving the transmission of tomato chlorotic dwarf viroid in eggplant. Annals of Applied Biology. 175(2):172-183. https://doi.org/10.1111/aab.12527S1721831752Bakker, D., Bruinsma, M., Dekter, R. W., Toonen, M. A. J., Verhoeven, J. T. J., & Koenraadt, H. M. S. (2015). Detection of PSTVd and TCDVd in seeds of tomato using real-time RT-PCR. EPPO Bulletin, 45(1), 14-21. doi:10.1111/epp.12195Barba, M., & James, D. (2017). Quarantine and Certification for Viroids and Viroid Diseases. Viroids and Satellites, 415-424. doi:10.1016/b978-0-12-801498-1.00039-5Broadbent, L. (1976). Epidemiology and Control of Tomato Mosaic Virus. Annual Review of Phytopathology, 14(1), 75-96. doi:10.1146/annurev.py.14.090176.000451Candresse, T., Marais, A., Tassus, X., Suhard, P., Renaudin, I., Leguay, A., … Blancard, D. (2010). First Report of Tomato chlorotic dwarf viroid in Tomato in France. Plant Disease, 94(5), 633-633. doi:10.1094/pdis-94-5-0633bCandresse, T., Verhoeven, J. T. J., Stancanelli, G., Hammond, R. W., & Winter, S. (2017). Other Pospiviroids Infecting Solanaceous Plants. Viroids and Satellites, 159-168. doi:10.1016/b978-0-12-801498-1.00015-2Červená, G., Nečekalová, J., Mikulková, H., Levkaničová, Z., Mertelík, J., Kloudová, K., … Ptáček, J. (2011). VIROIDS ON PETUNIA AND OTHER SOLANACEOUS CROPS IN THE CZECH REPUBLIC. Acta Horticulturae, (901), 35-40. doi:10.17660/actahortic.2011.901.3Constable F. &Moran J.(1996).PCR protocols for the detection of chrysanthemum stunt and potato spindle tuber viroids. Final Report for the Horticultural Research and Development Corporation Project number PT410. Victoria Australia: Department of Natural Resources and Environment.Daròs, J.-A. (2017). Eggplant Latent Viroid. Viroids and Satellites, 339-344. doi:10.1016/b978-0-12-801498-1.00032-2Di Serio, F., Flores, R., Verhoeven, J. T. J., Li, S.-F., Pallás, V., Randles, J. W., … Owens, R. A. (2014). Current status of viroid taxonomy. Archives of Virology, 159(12), 3467-3478. doi:10.1007/s00705-014-2200-6FAGOAGA, C., & DURAN-VILA, N. (1996). Naturally occurring variants of citrus exocortis viroid in vegetable crops. Plant Pathology, 45(1), 45-53. doi:10.1046/j.1365-3059.1996.d01-104.xFlores, R., Hernández, C., Alba, A. E. M. de, Daròs, J.-A., & Serio, F. D. (2005). Viroids and Viroid-Host Interactions. Annual Review of Phytopathology, 43(1), 117-139. doi:10.1146/annurev.phyto.43.040204.140243Fox, A., Daly, M., Nixon, T., Brurberg, M. B., Blystad, D.-R., Harju, V., … Adams, I. P. (2013). First report ofTomato chlorotic dwarf viroid(TCDVd) in tomato in Norway and subsequent eradication. New Disease Reports, 27, 8. doi:10.5197/j.2044-0588.2013.027.008Giguère, T., Raj Adkar-Purushothama, C., & Perreault, J.-P. (2014). Comprehensive Secondary Structure Elucidation of Four Genera of the Family Pospiviroidae. PLoS ONE, 9(6), e98655. doi:10.1371/journal.pone.0098655Giguère, T., & Perreault, J.-P. (2017). Classification of the Pospiviroidae based on their structural hallmarks. PLOS ONE, 12(8), e0182536. doi:10.1371/journal.pone.0182536Gruber, A. R., Lorenz, R., Bernhart, S. H., Neubock, R., & Hofacker, I. L. (2008). The Vienna RNA Websuite. Nucleic Acids Research, 36(Web Server), W70-W74. doi:10.1093/nar/gkn188Hadidi, A., Flores, R., Randles, J., & Semancik, J. (2003). Viroids. doi:10.1071/9780643069855Hailstones, D. L., Tesoriero, L. A., Terras, M. A., & Dephoff, C. (2003). Detection and eradication of Potato spindle tuber viroid in tomatoes in commercial production in New South Wales, Australia. Australasian Plant Pathology, 32(2), 317. doi:10.1071/ap03005Hammond, R. W. (2017). Economic Significance of Viroids in Vegetable and Field Crops. Viroids and Satellites, 5-13. doi:10.1016/b978-0-12-801498-1.00001-2Hammond, R. W. (2017). Seed, Pollen, and Insect Transmission of Viroids. Viroids and Satellites, 521-530. doi:10.1016/b978-0-12-801498-1.00048-6Hirakawa, H., Shirasawa, K., Miyatake, K., Nunome, T., Negoro, S., Ohyama, A., … Fukuoka, H. (2014). Draft Genome Sequence of Eggplant (Solanum melongena L.): the Representative Solanum Species Indigenous to the Old World. DNA Research, 21(6), 649-660. doi:10.1093/dnares/dsu027James, T., Mulholland, V., Jeffries, C., & Chard, J. (2008). First report of Tomato chlorotic dwarf viroid infecting commercial petunia stocks in the United Kingdom. Plant Pathology, 57(2), 400-400. doi:10.1111/j.1365-3059.2007.01727.xKeese, P., & Symons, R. H. (1985). Domains in viroids: evidence of intermolecular RNA rearrangements and their contribution to viroid evolution. Proceedings of the National Academy of Sciences, 82(14), 4582-4586. doi:10.1073/pnas.82.14.4582Kovalskaya, N., & Hammond, R. W. (2014). Molecular biology of viroid–host interactions and disease control strategies. Plant Science, 228, 48-60. doi:10.1016/j.plantsci.2014.05.006Kryczyński, S., Paduch-Cichal, E., & Skrzeczkowski, L. J. (1988). Transmission of Three Viroids Through Seed and Pollen of Tomato Plants. Journal of Phytopathology, 121(1), 51-57. doi:10.1111/j.1439-0434.1988.tb00952.xLi, R., Baysal-Gurel, F., Abdo, Z., Miller, S. A., & Ling, K.-S. (2015). Evaluation of disinfectants to prevent mechanical transmission of viruses and a viroid in greenhouse tomato production. Virology Journal, 12(1), 5. doi:10.1186/s12985-014-0237-5Ling, K.-S. (2017). Decontamination Measures to Prevent Mechanical Transmission of Viroids. Viroids and Satellites, 437-445. doi:10.1016/b978-0-12-801498-1.00041-3Ling, K.-S., Verhoeven, J. T. J., Singh, R. P., & Brown, J. K. (2009). First Report of Tomato chlorotic dwarf viroid in Greenhouse Tomatoes in Arizona. Plant Disease, 93(10), 1075-1075. doi:10.1094/pdis-93-10-1075bLing, K.-S., & Zhang, W. (2009). First Report of a Natural Infection by Mexican Papita Viroid and Tomato Chlorotic Dwarf Viroid on Greenhouse Tomatoes in Mexico. Plant Disease, 93(11), 1216-1216. doi:10.1094/pdis-93-11-1216aMackie, A. E., Coutts, B. A., Barbetti, M. J., Rodoni, B. C., McKirdy, S. J., & Jones, R. A. C. (2015). Potato spindle tuber viroid: Stability on Common Surfaces and Inactivation With Disinfectants. Plant Disease, 99(6), 770-775. doi:10.1094/pdis-09-14-0929-reMatsushita, Y., Kanda, A., Usugi, T., & Tsuda, S. (2008). First report of a Tomato chlorotic dwarf viroid disease on tomato plants in Japan. Journal of General Plant Pathology, 74(2), 182-184. doi:10.1007/s10327-008-0076-6Matsushita, Y., & Tsuda, S. (2014). Host ranges of Potato spindle tuber viroid, Tomato chlorotic dwarf viroid, Tomato apical stunt viroid, and Columnea latent viroid in horticultural plants. European Journal of Plant Pathology, 141(1), 193-197. doi:10.1007/s10658-014-0518-2Matsushita, Y., & Tsuda, S. (2016). Seed transmission of potato spindle tuber viroid, tomato chlorotic dwarf viroid, tomato apical stunt viroid, and Columnea latent viroid in horticultural plants. European Journal of Plant Pathology, 145(4), 1007-1011. doi:10.1007/s10658-016-0868-zMatsushita, Y., Usugi, T., & Tsuda, S. (2009). Host range and properties of Tomato chlorotic dwarf viroid. European Journal of Plant Pathology, 124(2), 349-352. doi:10.1007/s10658-008-9416-9Matsushita, Y., Usugi, T., & Tsuda, S. (2011). Distribution of tomato chlorotic dwarf viroid in floral organs of tomato. European Journal of Plant Pathology, 130(4), 441-447. doi:10.1007/s10658-011-9766-6Matsuura, S., Matsushita, Y., Kozuka, R., Shimizu, S., & Tsuda, S. (2009). Transmission of Tomato chlorotic dwarf viroid by bumblebees (Bombus ignitus) in tomato plants. European Journal of Plant Pathology, 126(1), 111-115. doi:10.1007/s10658-009-9515-2Matsuura, S., Matsushita, Y., Usugi, T., & Tsuda, S. (2010). Disinfection of Tomato chlorotic dwarf viroid by chemical and biological agents. Crop Protection, 29(10), 1157-1161. doi:10.1016/j.cropro.2010.05.018Minoia, S., Navarro, B., Delgado, S., Serio, F. D., & Flores, R. (2015). Viroid RNA turnover: characterization of the subgenomic RNAs of potato spindle tuber viroid accumulating in infected tissues provides insights into decay pathways operating in vivo. Nucleic Acids Research, 43(4), 2313-2325. doi:10.1093/nar/gkv034Nie, X. (2012). Analysis of Sequence Polymorphism and Population Structure of Tomato chlorotic dwarf viroid and Potato spindle tuber viroid in Viroid-Infected Tomato Plants. Viruses, 4(6), 940-953. doi:10.3390/v4060940Palukaitis, P. (1987). Potato spindle tuber viroid: Investigation of the long-distance, intra-plant transport route. Virology, 158(1), 239-241. doi:10.1016/0042-6822(87)90260-1PROHENS, J., SOLER, S., & NUEZ, F. (1999). The effects of thermotherapy and sodium hypochlorite treatments on pepino seed germination, a crucial step in breeding programmes. Annals of Applied Biology, 134(3), 299-305. doi:10.1111/j.1744-7348.1999.tb05268.xShiraishi, T., Maejima, K., Komatsu, K., Hashimoto, M., Okano, Y., Kitazawa, Y., … Namba, S. (2013). First report of tomato chlorotic dwarf viroid isolated from symptomless petunia plants (Petunia spp.) in Japan. Journal of General Plant Pathology, 79(3), 214-216. doi:10.1007/s10327-013-0444-8Sievers, F., Wilm, A., Dineen, D., Gibson, T. J., Karplus, K., Li, W., … Higgins, D. G. (2011). Fast, scalable generation of high‐quality protein multiple sequence alignments using Clustal Omega. Molecular Systems Biology, 7(1), 539. doi:10.1038/msb.2011.75Singh, R. P. (1992). Detection of Potato Spindle Tuber Viroid in the Pollen and Various Parts of Potato Plant Pollinated with Viroid-Infected Pollen. Plant Disease, 76(9), 951. doi:10.1094/pd-76-0951Singh, R. P., Nie, X., & Singh, M. (1999). Tomato chlorotic dwarf viroid: an evolutionary link in the origin of pospiviroids The GenBank accession number of the sequence reported in this paper is AF162131. Journal of General Virology, 80(11), 2823-2828. doi:10.1099/0022-1317-80-11-2823Singh, R. P., & Dilworth, A. D. (2008). Tomato chlorotic dwarf viroid in the ornamental plant Vinca minor and its transmission through tomato seed. European Journal of Plant Pathology, 123(1), 111-116. doi:10.1007/s10658-008-9344-8Singh, R. P., Dilworth, A. D., Ao, X., Singh, M., & Misra, S. (2009). Molecular and biological characterization of a severe isolate of Tomato chlorotic dwarf viroid containing a novel terminal right (TR) domain sequence. European Journal of Plant Pathology, 127(1), 63-72. doi:10.1007/s10658-009-9571-7Škorić, D. (2017). Viroid Biology. Viroids and Satellites, 53-61. doi:10.1016/b978-0-12-801498-1.00005-xVan Bogaert, N., Smagghe, G., Maes, M., De Backer, M., & De Jonghe, K. (2017). Phylogeny of five predominant pospiviroid species in Belgium. European Journal of Plant Pathology, 149(1), 25-33. doi:10.1007/s10658-017-1158-0Verhoeven, J. T. J., Jansen, C. C. C., Botermans, M., & Roenhorst, J. W. (2010). Epidemiological evidence that vegetatively propagated, solanaceous plant species act as sources ofPotato spindle tuber viroidinoculum for tomato. Plant Pathology, 59(1), 3-12. doi:10.1111/j.1365-3059.2009.02173.xVerhoeven, J. T. J., Hammond, R. W., & Stancanelli, G. (2017). Economic Significance of Viroids in Ornamental Crops. Viroids and Satellites, 27-38. doi:10.1016/b978-0-12-801498-1.00003-6Verhoeven, J. th. j., Jansen, C. C. C., Willemen, T. M., Kox, L. F. F., Owens, R. A., & Roenhorst, J. W. (2004). Natural infections of tomato by Citrus exocortis viroid, Columnea latent viroid, Potato spindle tuber viroid and Tomato chlorotic dwarf viroid. European Journal of Plant Pathology, 110(8), 823-831. doi:10.1007/s10658-004-2493-5Võ, T. T., Dehne, H.-W., & Hamacher, J. (2018). Transmission of Tomato chlorotic dwarf viroid by Myzus persicae assisted by Potato leafroll virus. Journal of Plant Diseases and Protection, 125(3), 259-266. doi:10.1007/s41348-018-0151-yWassenegger, M., Spieker, R. L., Thalmeir, S., Gast, F.-U., Riedel, L., & Sänger, H. L. (1996). A Single Nucleotide Substitution Converts Potato Spindle Tuber Viroid (PSTVd) from a Noninfectious to an Infectious RNA for Nicotiana tabacum. Virology, 226(2), 191-197. doi:10.1006/viro.1996.0646Yanagisawa, H., & Matsushita, Y. (2018). Differences in dynamics of horizontal transmission of Tomato planta macho viroid and Potato spindle tuber viroid after pollination with viroid-infected pollen. Virology, 516, 258-264. doi:10.1016/j.virol.2018.01.023Yanagisawa, H., Sano, T., Hase, S., & Matsushita, Y. (2019). Influence of the terminal left domain on horizontal and vertical transmissions of tomato planta macho viroid and potato spindle tuber viroid through pollen. Virology, 526, 22-31. doi:10.1016/j.virol.2018.09.02

Measurement of the cosmic ray spectrum above 4×10184{\times}10^{18} eV using inclined events detected with the Pierre Auger Observatory

A measurement of the cosmic-ray spectrum for energies exceeding $4{\times}10^{18}$ eV is presented, which is based on the analysis of showers with zenith angles greater than $60^{\circ}$ detected with the Pierre Auger Observatory between 1 January 2004 and 31 December 2013. The measured spectrum confirms a flux suppression at the highest energies. Above $5.3{\times}10^{18}$ eV, the "ankle", the flux can be described by a power law $E^{-\gamma}$ with index $\gamma=2.70 \pm 0.02 \,\text{(stat)} \pm 0.1\,\text{(sys)}$ followed by a smooth suppression region. For the energy ($E_\text{s}$) at which the spectral flux has fallen to one-half of its extrapolated value in the absence of suppression, we find $E_\text{s}=(5.12\pm0.25\,\text{(stat)}^{+1.0}_{-1.2}\,\text{(sys)}){\times}10^{19}$ eV.Comment: Replaced with published version. Added journal reference and DO