A low-cost Portable Device to Deliver Smoke, Volatile or Vaporized Substances to Drosophila melanogaster, Useful for Research and/or Educational Assays

Drosophila melanogaster has been used to test drugs of abuse, substances with potential benefits for medical purposes, as well as contaminants and hazardous volatile compounds. This model has also been used for the characterization of behavioral changes, physiopathological consequences, and subcellular mechanisms of the use of cocaine, methamphetamines, ethanol, nicotine, cannabinoids, toluene, and other airborne volatile organic compounds. When testing these substances, routes of administration are important to define. Admixing the test compounds with water or food is one suitable option in many cases, but the inhalation route is especially suitable when the administration of one or more volatile compounds is desired. One advantage of the administration of substances via the inhalation route is its rapid exchange and distribution throughout the cuticle and the tracheal system. In addition, this route allows treating a large group of individuals simultaneously. Moreover, the inhalation route is frequently used to administer different drugs to humans. A good model system shares physiology and molecular pathways with humans, and D. melanogaster possesses almost 75% homologous genes associated with human diseases. Methodologies to deliver the abovementioned substances usually include customized devices. Herein, we focus on the development of a low-cost customized device useful to deliver smoke or vaporizable compounds to D. melanogaster. This approach might be applied for acute or chronic exposure to vaporized substances. In particular, our device was utilized for testing cigarette smoke and vaporized cannabis extract on cardiac performance of adult individuals during chronic treatment. We are describing how to set up this low-cost portable device, useful for research and/or educational assays, taking advantage of the amenability of D. melanogaster to test different compounds in relatively short periods, and especially including a large number of individuals at the same time.Fil: Santalla, Manuela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Gómez, Ivana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Valverde, Carlos Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Ferrero, Paola Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional del Noroeste de la Provincia de Buenos Aires; Argentin

Long term behavior of the stirred vacuum on a Dirac chain: geometry blur and the random Slater ensemble

We characterize the long-term state of the 1D Dirac vacuum stirred by an impenetrable object, modeled as the ground state of a finite free-fermionic chain dynamically perturbed by a moving classical obstacle which suppresses the local hopping amplitudes. We find two different regimes, depending on the velocity of the obstacle. For a slow motion, the effective Floquet Hamiltonian presents features which are typical of the Gaussian orthogonal ensemble, and the occupation of the Floquet modes becomes roughly homogeneous. Moreover, the long term entanglement entropy of a contiguous block follows a Gaussian analogue of Page's law, i.e. a volumetric behavior. Indeed, the statistical properties of the reduced density matrices correspond to those of a random Slater determinant, which can be described using the Jacobi ensemble from random matrix theory. On the other hand, if the obstacle moves fast enough, the effective Floquet Hamiltonian presents a Poissonian behavior. The nature of the transition is clarified by the entanglement links, which determine the effective geometry underlying the entanglement structure, showing that the one-dimensionality of the physical Hamiltonian dissolves into a random adjacency matrix as we slow down the obstacle motion

Análisis de la relación entre el Yo-Yo Test y el consumo máximo de oxígeno en jóvenes jugadores de fútbol. [Analysis of the relationship between Yo-Yo Test and maximum oxygen uptake in young football players].

This work aimed to examine the validity of the Yo-Yo Endurance Test Level 1 to estimate the maximum oxygen uptake in football players. Participants were 15 male football players, ranging in age from 17 to 19 years old (M = 17,9; DT = 0,67), who developed a laboratory test until exhaustion and Yo-Yo Test Level 1. Results showed significant differences in the values directly and indirectly obtained, being the values lower in the oxygen uptake estimated with Yo-Yo Test. Furthermore, Yo-Yo Test performance was not related with maximum oxygen uptake relative to weight on laboratory, showing that Yo-Yo Endurance Test Level 1, through the equation described by Bangsbo, Iaia, and Krustrup (2008) is not a valid test to estimate maximum oxygen uptake in young football players

Combined HIIT and Resistance Training in Very Long-Chain Acyl-CoA Dehydrogenase Deficiency: A Case Report

Very long-chain acyl-CoA dehydrogenase deficiency (VLCADD) is a rare disorder of mitochondrial fatty acid β-oxidation characterized by a spectrum of clinical manifestations. Patients with the adult-onset form can present with muscle pain, rhabdomyolysis and myoglobinuria after physiological stress, such as fasting and exercise. We report on a 23-year-old female patient with a history of recurrent rhabdomyolysis. The patient completed a 6-month supervised combined (high-intensity interval training [HIIT] + resistance training) program, with the addition of a medium chain triglyceride + carbohydrate supplement provided 60 min before each session. The HIIT consisted of 6 sets of 70–80 s performed at maximum intensity with a minimum cadence of 100 rpm. Resistance training consisted of a circuit of basic exercises with dumbbells and elastic bands, with sets of 4–7 repetitions. The patient was evaluated at months 0, 3 and 6 using an incremental discontinuous step protocol, with steps of 1 min of exercise/1 min of passive recovery, at a high pedal cadence. The test started at 10 W, with a load increase of 10 W/step. Blood creatine kinase (CK) concentration was measured before each evaluation. There was a training-induced increment of 90.2% in peak oxygen uptake (VO2peak), 71.4% in peak power output and 24.7% in peak heart rate. The patient reported no muscle pain, contractures, rhabdomyolysis (basal CK concentration was always <200 U/L) or hospital admissions during the training period. After completion of 6-month program, the patient remained active, doing similar but non-supervised training for 1.5 years (to date). During this period, the patient has not reported myalgias, contractures, rhabdomyolysis or hospital admissions. Our preliminary data suggest that it is possible to carry out a combined (HIIT + strength) training program in patients with VLCADD, safely (without muscle contractures or rhabdomyolysis) and obtaining high values of VO2peak and cycling power output

The influence of the floater position on the load of soccer players during a 4 vs 4 + 2 game

The purpose of this study was to examine the load of regular players and floater players in a specific small-sided game. Twenty semi-professional soccer players performed one 4 vs 4+2, modifying the position of the floaters (internal, external, zone, square and without floaters). Total distance covered, distance covered at speeds between 7-13.9 km·h−1, 14-17.9 km·h−1, and ≥18 km·h−1, accelerations and decelerations between 2.5-4 m·s−2, maximal and mean heart rate (HR) and rating of perceived exertion (RPE) were analyzed. Internal floaters achieved greater total distance covered, accelerations and RPE than in any other position. Internal and external floaters achieved more distance covered at 7-13.9, 14-17.9, >18 km·h-1 and HR than zone and square floaters. With internal floaters, regular players covered more distance covered >18 km·h-1 than in any other 4vs4+2 format, and with internal and external floaters regular players covered greater total distance and distance at 14-17.9 km·h-1 than without floaters or with zone or square floaters. Regular players showed greater total distance covered, distance covered at 7-13.9, 14-17.9, >18 km·h-1, accelerations and RPE than floaters in all 4vs4+2. These data showed that floater position in 4vs4+2 game influenced the internal and external load of both the regular players and the floaters

The Second Wind in McArdle Patients: Fitness Matters

Glucogenosis tipo 5; Deficiencia de miofosforilasa; Umbral de ventilaciónGlycogenosis type 5; Myophosphorylase deficiency; Ventilatory thresholdGlucogenosi tipus 5; Deficiència de miofosforilasa; Llindar ventilatoriBackground: The “second wind” (SW) phenomenon—commonly referring to both an initial period of marked intolerance to dynamic exercise (e.g., brisk walking) that is not followed by perceived improvement and disappearance of previous tachycardia (i.e., the actual “SW”) until 6–10 min has elapsed—is an almost unique feature of McArdle disease that limits adherence to an active lifestyle. In this regard, an increase in the workload eliciting the SW could potentially translate into an improved patients’ exercise tolerance in daily life. We aimed to determine whether aerobic fitness and physical activity (PA) levels are correlated with the minimum workload eliciting the SW in McArdle patients—as well as with the corresponding heart rate value. We also compared the SW variables and aerobic fitness indicators in inactive vs. active patients. Methods: Fifty-four McArdle patients (24 women, mean ± SD age 33 ± 12 years) performed 12-min constant-load and maximum ramp-like cycle-ergometer tests for SW detection and aerobic fitness [peak oxygen uptake (VO2peak) and workload and ventilatory threshold] determination, respectively. They were categorized as physically active/inactive during the prior 6 months (active = reporting ≥150 min/week or ≥75 min/week in moderate or vigorous-intensity aerobic PA, respectively) and were also asked on their self-report of the SW. Results: Both peak and submaximal indicators of aerobic fitness obtained in the ramp tests were significantly correlated with the workload of the SW test, with a particularly strong correlation for the VO2peak and peak workload attained by the patients (both Pearson’s coefficients > 0.70). Twenty (seven women) and 24 patients (18 women) were categorized as physically active and inactive, respectively. Not only the aerobic fitness level [∼18–19% higher values of VO2peak (ml⋅kg–1⋅min–1)] but also the workload of the SW tests was significantly higher in active than in inactive patients. All the inactive patients reported that they experienced the SW during walking/brisk walking in daily life, whereas active patients only reported experiencing this phenomenon during more strenuous activities (very brisk walking/jogging and bicycling). Conclusion: A higher aerobic fitness and an active lifestyle are associated with a higher workload eliciting the so-called SW phenomenon in patients with McArdle disease, which has a positive impact on their exercise tolerance during daily living.Research by AL, CF-L, GN-G, MM, and TP was funded by the Instituto de Salud Carlos III (ISCIII) and Fondos Feder (grants PI18/00139, PI20/00645, PI18/00713, PI17/00093, and PI19/01313). CF-L and GN-G were supported by Miguel Servet contracts (CP18/00034 and CPII19/00021, respectively) from ISCIII. The funders had no role in study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the manuscript for publication

Smoking flies: Testing the effect of tobacco cigarettes on heart function of Drosophila melanogaster

Studies about the relationship between substances consumed by humans and their impact on health, in animal models, have been a challenge due to differences between species in the animal kingdom. However, the homology of certain genes has allowed extrapolation of certain knowledge obtained in animals. Drosophila melanogaster, studied for decades, has been widely used as model for human diseases as well as to study responses associated with the consumption of several substances. In the present work we explore the impact of tobacco consumption on a model of 'smoking flies'. Throughout these experiments, we aim to provide information about the effects of tobacco consumption on cardiac physiology. We assessed intracellular calcium handling, a phenomenon underlying cardiac contraction and relaxation. Flies chronically exposed to tobacco smoke exhibited an increased heart rate and alterations in the dynamics of the transient increase of intracellular calcium in myocardial cells. These effects were also evident under acute exposure to nicotine of the heart, in a semi-intact preparation. Moreover, the alpha 1 and 7 subunits of the nicotinic receptors are involved in the heart response to tobacco and nicotine under chronic (in the intact fly) as well as acute exposure (in the semi-intact preparation). The present data elucidate the implication of the intracellular cardiac pathways affected by nicotine on the heart tissue. Based on the probed genetic and physiological similarity between the fly and human heart, cardiac effects exerted by tobacco smoke in Drosophila advances our understanding of the impact of it in the human heart. Additionally, it may also provide information on how nicotine-like substances, e.g. neonicotinoids used as insecticides, affect cardiac function.Fil: Santalla, Manuela. Universidad Nacional del Noroeste de la Provincia de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Pagola, Lucia Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Gómez, Ivana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Balcazar, Dario Emmanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Valverde, Carlos Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Ferrero, Paola Viviana. Universidad Nacional del Noroeste de la Provincia de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentin

Aging and CaMKII alter intracellular Ca2+ transients and heart rhythm in Drosophila melanogaster

Aging is associated to disrupted contractility and rhythmicity, among other cardiovascular alterations. Drosophila melanogaster shows a pattern of aging similar to human beings and recapitulates the arrhythmogenic conditions found in the human heart. Moreover, the kinase CaMKII has been characterized as an important regulator of heart function and an arrhythmogenic molecule that participate in Ca2+ handling. Using a genetically engineered expressed Ca2+ indicator, we report changes in cardiac Ca 2+ handling at two different ages. Aging prolonged relaxation, reduced spontaneous heart rate (HR) and increased the occurrence of arrhythmias, ectopic beats and asystoles. Alignment between Drosophila melanogaster and human CaMKII showed a high degree of conservation and indicates that relevant phosphorylation sites in humans are also present in the fruit fly. Inhibition of CaMKII by KN-93 (CaMKII-specific inhibitor), reduced HR without significant changes in other parameters. By contrast, overexpression of CaMKII increased HR and reduced arrhythmias. Moreover, it increased fluorescence amplitude, maximal rate of rise of fluorescence and reduced time to peak fluorescence. These results suggest that CaMKII in Drosophila melanogaster acts directly on heart function and that increasing CaMKII expression levels could be beneficial to improve contractility.Centro de Investigaciones CardiovascularesCentro de Investigaciones Inmunológicas Básicas y Aplicada

Muscle glycogen unavailability and fat oxidation rate during exercise: Insights from McArdle disease

Glycogen store disease; Muscle fatigue; Substrate oxidationEnfermedad de almacenamiento de glucógeno; Fatiga muscular; Oxidación del sustratoMalaltia d'emmagatzematge de glucogen; Fatiga muscular; Oxidació del substratCarbohydrate availability affects fat metabolism during exercise; however, the effects of complete muscle glycogen unavailability on maximal fat oxidation (MFO) rate remain unknown. Our purpose was to examine the MFO rate in patients with McArdle disease, comprising an inherited condition caused by complete blockade of muscle glycogen metabolism, compared to healthy controls. Nine patients (three women, aged 36 ± 12 years) and 12 healthy controls (four women, aged 40 ± 13 years) were studied. Several molecular markers of lipid transport/metabolism were also determined in skeletal muscle (gastrocnemius) and white adipose tissue of McArdle (Pygm p.50R∗/p.50R∗) and wild-type male mice. Peak oxygen uptake (V˙O2peak), MFO rate, the exercise intensity eliciting MFO rate (FATmax) and the MFO rate-associated workload were determined by indirect calorimetry during an incremental cycle-ergometer test. Despite having a much lower V˙O2peak (24.7 ± 4 vs. 42.5 ± 11.4 mL kg−1 min−1, respectively; P < 0.0001), patients showed considerably higher values for the MFO rate (0.53 ± 0.12 vs. 0.33 ± 0.10 g min−1, P = 0.001), and for the FATmax (94.4 ± 7.2 vs. 41.3 ± 9.1 % of V˙O2peak, P < 0.0001) and MFO rate-associated workload (1.33 ± 0.35 vs. 0.81 ± 0.54 W kg−1, P = 0.020) than controls. No between-group differences were found overall in molecular markers of lipid transport/metabolism in mice. In summary, patients with McArdle disease show an exceptionally high MFO rate, which they attained at near-maximal exercise capacity. Pending more mechanistic explanations, these findings support the influence of glycogen availability on MFO rate and suggest that these patients develop a unique fat oxidation capacity, possibly as an adaptation to compensate for the inherited blockade in glycogen metabolism, and point to MFO rate as a potential limiting factor of exercise tolerance in this disease.Research by the IA and CR-L group is funded by the Biomedical Research Networking Centre on Frailty and Healthy Aging (CIBERFES, CB16/10/00314 and CB16/10/00477). IR-G is supported by a postdoctoral contract from Universidad de Castilla–La Mancha (2021/5937). PLV is supported by a Sara Borrell contract from Instituto de Salud Carlos III (CD21/00138). Research by AL and TP is funded by the Spanish Ministry of Economy and Competitiveness and Fondos FEDER (PI18/00139 and PI19/01313, respectively)

Aging and CaMKII alter intracellular Ca2+ transients and heart rhythm in Drosophila Melanogaster

Aging is associated to disrupted contractility and rhythmicity, among other cardiovascular alterations. Drosophila melanogaster shows a pattern of aging similar to human beings and recapitulates the arrhythmogenic conditions found in the human heart. Moreover, the kinase CaMKII has been characterized as an important regulator of heart function and an arrhythmogenic molecule that participate in Ca2+ handling. Using a genetically engineered expressed Ca2+ indicator, we report changes in cardiac Ca2+ handling at two different ages. Aging prolonged relaxation, reduced spontaneous heart rate (HR) and increased the occurrence of arrhythmias, ectopic beats and asystoles. Alignment between Drosophila melanogaster and human CaMKII showed a high degree of conservation and indicates that relevant phosphorylation sites in humans are also present in the fruit fly. Inhibition of CaMKII by KN-93 (CaMKII-specific inhibitor), reduced HR without significant changes in other parameters. By contrast, overexpression of CaMKII increased HR and reduced arrhythmias. Moreover, it increased fluorescence amplitude, maximal rate of rise of fluorescence and reduced time to peak fluorescence. These results suggest that CaMKII in Drosophila melanogaster acts directly on heart function and that increasing CaMKII expression levels could be beneficial to improve contractility.Fil: Santalla, Manuela. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Departamento de Ciencias Básicas y Experimentales; ArgentinaFil: Valverde, Carlos Alfredo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; ArgentinaFil: Harnichar, Alejandro Ezequiel. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; ArgentinaFil: Lacunza, Ezequiel. Universidad Nacional de La Plata. Facultad de Ciencias Medicas. Centro de Investigaciones Inmunologicas Basicas y Aplicadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Aguilar Fuentes, Javier. Universidad Autónoma de Chiapas; MéxicoFil: Mattiazzi, Ramona Alicia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; ArgentinaFil: Ferrero, Paola Viviana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Departamento de Ciencias Básicas y Experimentales; Argentin