Efeitos do Óleo de Copaíba em Marcadores Periféricos de Estresse Oxidativo em um Modelo de Cor Pulmonale em Ratos / Effects of Copaiba Oil in Peripheral Markers of Oxidative Stress in a Model of Cor Pulmonale in Rats

Resumo Fundamento Até o presente momento, os efeitos sistêmicos do óleo de copaíba jamais foram documentados no Cor pulmonale induzido por monocrotalina. Objetivos Investigar os efeitos do óleo de copaíba nos marcadores periféricos de stress oxidativo em ratos com Cor pulmonale. Métodos Ratos Wistar machos (170±20g, n=7/grupo) foram divididos em quatro grupos: controle (CO), monocrotalina (MCT), óleo de copaíba (O), e monocrotalina + óleo de copaíba (MCT-O). Foi administrada a MCT (60 mg/kg i.p.) e, depois de uma semana, foi iniciado o tratamento com óleo de copaíba (400 mg/kg/day-gavagem-14 dias). Foi realizado o ecocardiograma e, depois disso, foi coletado sangue do tronco para a realização de avaliações de stress oxidativo. Análise estatística: ANOVA de duas vias com teste Student-Newman-Keuls post hoc. P-valores <0,05 foram considerados significativos. Resultados O óleo de copaíba reduziu a resistência vascular pulmonar e a hipertrofia do ventrículo direito (VD) hipertrofia (Índice de Fulton (mg/mg)): MCT-O= 0,39±0,03; MCT= 0,49±0,01), e função sistólica melhorada (fração de encurtamento do VD, %) no grupo MCT-O (17,8±8,2) em comparação com o grupo de MCT (9,4±3,1; p<0,05). Além disso, no grupo MCT-O, espécies reativas do oxigênio e os níveis de carbonila foram reduzidos, e os parâmetros antioxidantes aumentaram no sangue periférico (p <0,05). Conclusões Os resultados deste estudo sugerem que o óleo de copaíba tem um efeito antioxidante sistêmico interessante, que se reflete na melhoria da função e na morfometria do VD nesse modelo de Cor pulmonale . A atenuação do Cor pulmonale promovida pelo óleo de copaíba coincidiu com uma redução no stress oxidativo sistêmico.

Abstract Background To date, copaiba oil's systemic effects have never documented in Cor pulmonale induced by monocrotaline. Objectives To investigate copaiba oil's effects in peripheral markers of oxidative stress in rats with Cor pulmonale. Methods Male Wistar rats (170±20g, n=7/group) were divided into four groups: control (CO), monocrotaline (MCT), copaiba oil (O), and monocrotaline+copaiba oil (MCT-O). MCT (60 mg/kg i.p.) was administered, and after one week, treatment with copaiba oil (400 mg/kg/day-gavage-14 days) was begun. Echocardiography was performed and, later, trunk blood collection was performed for oxidative stress evaluations. Statistical analysis: two-way ANOVA with Student-Newman-Keuls post-hoc test. P values<0.05 were considered significant. Results Copaiba oil reduced pulmonary vascular resistance and right ventricle (RV) hypertrophy (Fulton index (mg/mg): MCT-O=0.39±0.03; MCT=0.49±0.01), and improved RV systolic function (RV shortening fraction, %) in the MCT-O group (17.8±8.2) as compared to the MCT group (9.4±3.1; p<0.05). Moreover, in the MCT-O group, reactive oxygen species and carbonyl levels were reduced, and antioxidant parameters were increased in the peripheral blood (p<0.05). Conclusions: Our results suggest that copaiba oil has an interesting systemic antioxidant effect, which is reflected in the improvements in function and RV morphometry in this Cor pulmonale model. Cor pulmonale attenuation promoted by copaiba oil coincided with a reduction in systemic oxidative stress.

Investigación traslacional en microbioma (InTraMic): un área de interés común y creciente en el IMTIB, Hospital Italiano e Instituto Universitario / Translational research in microbiome (InTraMic): an area of common and growing interest at IMTIB, Hospital Italiano and Instituto Universitario

Se estima que aproximadamente 100 trillones de microorganismos (incluidos bacterias, virus y hongos) residen en el intestino humano adulto y que el total del material genético del microbioma es 100 veces superior al del genoma humano. Esta comunidad, conocida como microbioma se adquiere al momento del nacimiento a través de la flora comensal de la piel, vagina y heces de la madre y se mantiene relativamente estable a partir de los dos años desempeñando un papel crítico tanto en el estado de salud como en la enfermedad. El desarrollo de nuevas tecnologías, como los secuenciadores de próxima generación (NGS), permiten actualmente realizar un estudio mucho más preciso de ella que en décadas pasadas cuando se limitaba a su cultivo. Si bien esto ha llevado a un crecimiento exponencial en las publicaciones, los datos sobre las poblaciones Latinoamérica son casi inexistentes. La investigación traslacional en microbioma (InTraMic) es una de las líneas que se desarrollan en el Instituto de Medicina Traslacional e Ingeniería Biomédica (IMTIB). Esta se inició en 2018 con la línea de cáncer colorrectal (CCR) en una colaboración con el Colorectal Cancer Research Group del Leeds Institute of Medical Research en el proyecto Large bowel microbiome disease network: Creation of a proof of principle exemplar in colorectal cancer across three continents. A fines de 2019 se cumplió el objetivo de comprobar la factibilidad de la recolección, envío y análisis de muestras de MBF en 5 continentes, incluyendo muestras provenientes de la Argentina, Chile, India y Vietnam. Luego de haber participado de capacitaciones en Inglaterra, se ha cumplido con el objetivo de la etapa piloto, logrando efectivizar la recolección, envío y análisis metagenómico a partir de la secuenciación de la región V4 del ARNr 16S. En 2019, la línea de enfermedad de hígado graso no alcohólico se sumó a la InTraMic iniciando una caracterización piloto en el marco de una colaboración con el laboratorio Novartis. Los resultados de ese estudio, así como el de cáncer colorrectal, están siendo enviados a publicación. En 2020, con la incorporación de la línea de trasplante alogénico de células progenitoras hematopoyéticas, fue presentado un proyecto para un subsidio del CONICET que ha superado la primera etapa de evaluación. En el presente artículo se brinda una actualización sobre la caracterización taxonómica de microbioma y se describen las líneas de investigación en curso. (AU)

It is estimated that approximately 100 trillion microorganisms (including bacteria, viruses, and fungi) reside in the adult human intestine, and that the total genetic material of the microbiome is 100 times greater than that of the human genome. This community, known as the microbiome, is acquired at birth through the commensal flora of the mother's skin, vagina, and feces and remains relatively stable after two years, playing a critical role in both the state of health and in disease. The development of new technologies, such as next-generation sequencers (NGS), currently allow for a much more precise study of it than in past decades when it was limited to cultivation. Although this has led to exponential growth in publications, data on Latin American populations is almost non-existent. Translational research in microbiome (InTraMic) is one of the lines developed at the Instituto de Medicina Traslacional e Ingeniería Biomédica (IMTIB). This started in 2018 with the Colorectal Cancer Line (CRC) in a collaboration with the Colorectal Cancer Research Group of the Leeds Institute of Medical Research in the project "Large bowel microbiome disease network: Creation of a proof of principle exemplar in colorectal cancer across three continents". At the end of 2019, the objective of verifying the feasibility of collecting, sending and analyzing MBF samples on 5 continents, including samples from Argentina, Chile, India and Vietnam, was met. After having participated in training in England, the objective of the pilot stage has been met, achieving the collection, delivery and metagenomic analysis from the sequencing of the V4 region of the 16S rRNA. In 2019, the non-alcoholic fatty liver disease line joined InTraMic, initiating a pilot characterization in the framework of a collaboration with the Novartis laboratory. The results of that study, as well as that of colorectal cancer, are being published. In 2020, with the incorporation of the allogeneic hematopoietic stem cell transplantation line, a project was presented for a grant from the CONICET that has passed the first stage of evaluation. This article provides an update on the taxonomic characterization of the microbiome and describes the lines of ongoing research. (AU)

Análisis de polifenoles, actividad antioxidante y genotoxicidad en especies argentinas de Lippia y Aloysia (Verbenaceae) / Survey on polyphenols, antioxidant activity and genotoxicity on Argentinean species of Lippia and Aloysia (Verbenaceae)

Three species and a variety from the Verbenaceae belonging to the argentinean flora were analyzed: Aloysia gratissima var. gratissima (Gill. Et Hook) Tronc., Aloysia gratissima var. schulziana (Moldenke) Botta, Aloysia polystachya (Griseb.) Moldenke and Lippia integrifolia (Griseb.) Hieron., the first two known as “cedrón del monte”, the third as “yerba del burro” and the other as “incayuyo”. Their leaves are used as digestive in folk medicine. The research was performed on quantification of polyphenols (total phenols, total flavonoids and total hydroxicinnamic acids) and on determination of antioxidant and genotoxic activities. L. integrifolia showed the highest total phenol concentration and A. polystachya the lowest, but with the highest concentration of hydroxicinnamic acids. When flavonoids were quantified, A. polystachya showed the lowest concentations and A. gratissima var. schultziana the highest. According to quantification of total polyphenols, the lowest antioxidant activity was measured in A. polystachya extracts, followed by those from A. gratissima var. gratissima and A. gratissima var. schulziana, while extracts from L. integrifolia showed the highest antioxidant activity. No genotoxic activity was detected by means of the comet-assay for any of the species. This absence of genotoxicity, added to the relevant antioxidant activity, could justify the use of leaves infusions and decoctions from these species in treatments of those pathologies where antioxidants play an important role. These studies suggest their sure use in folk medicine, in which people include them among their medicinal plants.

Se analizaron 3 especies y una variedad de la familia Verbenaceae de la flora argentina: Aloysia gratissima var. gratissima (Gill. Et Hook) Tronc., Aloysia gratissima var. schulziana (Moldenke) Botta, Aloysia polystachya (Griseb.) Moldenke y Lippia integrifolia (Griseb.) Hieron., conocidas como “cedrón del monte” las dos primeras, “yerba del burro” la tercera e “incayuyo” la restante. De todas ellas se emplean las hojas en la medicina popular como digestivas. Se investigaron los contenidos de polifenoles (fenoles totales, flavonoides totales y ácidos hidroxicinámicos totales) y se determinó la capacidad antioxidante y genotoxicidad de las preparaciones acuosas (infusiones y cocimientos). L. integrifolia fue la especie con mayor contenido de fenoles totales y A. polystachya la menor. En el análisis de los hidroxicinámicos, A. polystachya fue la que mayor concentración mostró. En cuanto a los flavonoides, A. polystachya fue la que poseyó menor concentración en tanto que A. gratissima var. schultziana fue la más rica en estos compuestos. A. polystachya es la especie que presenta los menores valores de actividad antioxidante, seguida por A. gratissima var. gratissima y A. gratissima var. schulziana, mientras que L. integrifolia es la especie con mayor actividad antioxidante, en correlación con los valores de polifenoles totales. Ninguna de las especies analizadas demostró actividad genotóxica en el ensayo del cometa. La ausencia de genotoxicidad, sumada a una marcada actividad antioxidante, justificarían el empleo de las infusiones y cocimientos en el tratamiento de aquellas patologías donde los agentes antioxidantes juegan un importante rol terapéutico. Estos estudios sugieren el empleo seguro en la medicina tradicional de los pueblos y comunidades que las incluyen entre sus plantas medicinales.

Potencial antioxidativo da lágrima de adultos jovens / Tear antioxidant potential in young adults

OBJETIVO: Estudar os antioxidantes da lágrima humana, estimulada pelo corte de cebola e a possível influência dos hábitos de vida sobre estas medidas. MÉTODOS: A amostra consistiu de dez adultos jovens, que responderam questionário sobre o hábito de fumar, de ingerir bebidas alcoólicas, frutas, vegetais e cereais, de usar vitaminas e/ou drogas e de praticar exercícios. O potencial reativo antioxidante total (TRAP) foi analisado por meio da quimioluminescência do luminol, a superóxido dismutase (SOD) foi medida pela inibição do piragolol e a absorbância de H2O2 a 240 nm foi utilizada para identificar a catalase. RESULTADOS: A média ± DP dos valores de potencial reativo antioxidante total foi 33,8±11,5 µM e de superóxido dismutase foi 10,8±1,9 U/mL. Não foi identificada atividade da catalase. Detectou-se associação entre a prática regular de exercícios e aumento nos valores de potencial reativo antioxidante total (p=0,021), com diferença de 18,6 µM entre os indivíduos que se exercitavam pelo menos uma vez por semana e os sedentários. Sexo masculino e aumento na concentração de potencial reativo antioxidante total também se associaram estatisticamente (p=0,013), com diferença de 16,3 µM entre os sexos. Houve associação entre hábito tabágico e aumento na superóxido dismutase (p=0,041), com diferença de 3,3 U/mL entre fumantes de mais de cinco cigarros/dia e não fumantes. O uso de vitamina C também demonstrou associação com a superóxido dismutase (p=0,018); a diferença para os que tomavam vitamina C foi de 3,3 U/mL a mais. CONCLUSÃO: Os antioxidantes do lacrimejamento reflexo foram mensuráveis em adultos jovens, e diferentes variáveis parecem ter influenciado nos resultados.

PURPOSE: To study the antioxidant status of human tears, stimulated by onion fumes and the possible influence of the life habits thereon were measured. METHODS: Subjects were ten healthy young adults, who answered a questionnaire about smoking, alcohol ingestion, fruit, vegetable, cereal, and vitamin intake and/or intake of other drugs, and physical exercise habits. Chemoluminescensce of luminol was used to analyze the total reactive antioxidant potential (TRAP), inhibition of piragollol was used to measure superoxide dismutase (SOD) and absorbance of H2O2 at 240 nm was used to identify catalase. RESULTS: Mean ± SD value for total reactive antioxidant potential was 33.8±11.5 µM and for superoxide dismutase 10.8±1.9 U/mL. Catalase was not identified. Regular exercise was associated with increased total reactive antioxidant potential values (p=0.021), with a difference of 18.6 µM between individuals who exercise at least once a week and sedentary individuals. Male gender and total reactive antioxidant potential values were statistically associated (p=0.013), with a difference of 16.3 µM between genders. There was an association between smoking and increased superoxide dismutase values (p=0.041), with a difference of 3.3 U/mL between smokers of more than five cigarettes/day and non-smokers. Also, vitamin C intake and superoxide dismutase values were associated (p=0.018); the difference for vitamin C takers was 3.3 U/mL. CONCLUSION: Reflex tearing antioxidants were measurable in healthy young adults, and different variables apparently influenced their values.

Alterações gastrointestinais no diabetes mellitus: estresse oxidativo e fluxo sangüíneo da artéria mesentérica - estudo experimental / Gastrointestinal alterations in diabetes mellitus: oxidative stress and blood flow in the superior mesenteric artery - experimental study

RACIONAL: Os sintomas gastrointestinais são freqüentes no diabetes mellitus e podem estar relacionados com o estresse oxidativo, que é definido pelo desequilíbrio entre os sistemas pró-oxidante e o antioxidante. OBJETIVO: Avaliar algumas das alterações gastrointestinais no modelo de diabetes mellitus, como o estresse oxidativo no estômago e no fígado de animais diabéticos e o fluxo sangüíneo na artéria mesentérica superior em diferentes tempos de estudo. MATERIAL E MÉTODOS: Os parâmetros utilizados para verificar o estresse oxidativo no fígado e no estômago foram a mensuração da lipoperoxidação, através das técnicas das substâncias reativas ao ácido tiobarbitúrico e da quimiluminescência e a avaliação da atividade das enzimas antioxidantes catalase, superóxido dismutase e glutationa transferase. Utilizaram-se ratos machos Wistar, pesando entre 250-350 g, que foram divididos em quatro grupos: grupo I - 7 dias de diabetes, grupo II- 30 dias de diabetes, grupo III - 60 dias de diabetes e grupo IV - 90 dias de diabetes. O diabetes foi induzido por administração de estreptozotocina 70 mg/kg intraperitonialmente. RESULTADOS: Houve aumento significativo na lipoperoxidação no estômago e no fígado de animais diabéticos somente no tempo de 90 dias. No estômago, foi encontrada significativa diminuição na atividade das enzimas antioxidantes catalase e glutationa transferase. No fígado, somente a enzima glutationa transferase apresentou diminuição significativa. Houve aumento no fluxo da artéria mesentérica superior dos animais diabéticos com 90 dias, quando comparados aos animais-controle. CONCLUSÕES: É possível supor que o aumento no estresse oxidativo no estômago e no fígado e a alteração no fluxo sangüíneo da artéria mesentérica superior sejam influenciados pelo tempo de diabetes e pela hiperglicemia encontrada nos animais estudados, o que determinaria as alterações gastrointestinais.

Antioxidant enzymes and their modification under oxidative stress conditions

There are many cellular defense strategies against processes mediated by active oxygen species, including scavenger molecules and the enzymes catalase, superoxide dismutase and glutathione peroxidase. The activity of these antioxidant enzymes allows to keep O2- and H2O2 steady state concentrations at low levels, compatible with cell function. In most pathologies considered, modifications in levels of enzymatic activity have been observed in relation to values of control patients. Both increased and decreased levels have been found and in almost all systems the response of the three enzymes has been parallel. Different models of chronic treatment have also been considered. Parallel variations could be noted again regarding the antioxidant activity of these three enzymes. Moreover, there exists a correlation between the increase in the enzymatic activities and the acquired protection by treatment with barbital as reflected by the measured values of spontaneous chemiluminescence. on the other hand, increased levels of chemiluminescence in liver homogenates and diminished enzymatic activities were found in other experimental models. The response to these treatments is not the same in different organs since the rates of production of active oxygen species as well as the antioxidant defense activities are different in each organ. Finally, data of acute models of oxidative stress were compiled. Some of them have shown a biphasic response. At first a decrease in levels of enzymatic activities could be seen in response to the injury. Afterwards, an increase in the activity of the antioxidant enzymes followed as a consequence of an enzymatic induction or activation.

Hydrogen peroxide as a tool for studying oxidative stress in the heart

Hydrogen peroxide (H2O2) was observed to depolarize the frog sartorius muscle and promote rhythmic contraction of frog cardiac ventricular rings or their contracture. This last effect was sodium-dependent. H2O2 perfused or injected into the aorta of the isolated rat heart induces a positive inotropic effect (with cardiac arrhythmias such as extrasystolic potentiations) followed by deoression of contractility or cardiac contractures, according to the dose employed. The last effects is similar to the "stone heart"observed in the reperfusion injury and may be ascribed to lipoperoxidation (LPO) of the membrane lipids, to protein damage, to reduction in the ATP level and/or to cardioactive compounds liberated by LPO. Besides its direct effect on the ATP level, H2O2 would react with iron ions to produce hydroxyl radicals that attack the cellular membranes. Deferoxamine, an iron chelator and scavenger of hydroxyl radicals, reduced the contractures induced by H2O2. Perfusion with H2O2 increased the LPO of cardiac homogenates measured by chemiluminescence, oxygen uptake and malonaldehyde formation. The fall in ATP levels and the LPO would result in calcium overload of the cardiac fibers and contracture ("stone heart"). The 45Ca uptake was increased by incubation of cardiac strips with H2O2. Previous perfusion of the isolated rat heart with nifedipine or indomethacin reduced the H2O2 cardiac contracture. Vitamin A, a quencher of singlet oxygen liberated during LPO, reduces the H2O2 cardiac contractures and also LPO. Gradual physical exercises, besides increasing the oxygen consumption, protected the heart from oxidative stress. The experimental production of hypothyroidism protected the heart against the H2O2 oxidative stress. The hearts of rats submitted to hypertension with high renin levels showed increased LPO, measured by chemiluminescence and oxygen uptake, indicating that this condition may be produced by oxygen species or causes their production. All these findings give support to the idea that the ischemia-reperfusion injury is an active oxygen species associated disorder that induces cardiac stiffness or contractures that would be produced by calcium overload. Thus, H2O2 may be useful for inducing experimental oxidative stress in the heart and for studying its oxidative status in physiological and pathological situations.

Estudio comparativo de tres soluciones cardiopléjicas en pacientes sometidos a cirugía cardíaca / Comparative study of three cardioplegic solutions in patients submitted to cardiac surgery

Veinte pacientes operadores de cirugía de by pass aortocoronario fueron divididos en grupo I (n=7), que recibió como protección miocárdica una solución cardiopléjica standard; grupo II (n=6), que recibió una solución cardiopléjica conteniendo manitol; y grupo III (n=7) que recibió solución cardiopléjica con deferoxamina. Se tomaron biopsias de miocardio, previo al período isquémico (muestras A o preisquémicas) y a los 10 minutos de la reperfusión (muestras B o de reperfusión) las cuales fueron procesadas para quimioluminiscencia para detectar actividad de radicales libres y para microscopía electrónica con el objeto de evaluar lesión miocárdica a nivel de ultraestructura. Se observó que en el grupo I se producía un significativo aumento de los valores de quimioluminiscencia en las muestras B y ésto se asociaba con la presencia de áreas con franco edema mitocondrial. En los grupos II y III no había diferencia significativa entre las muestras A y B. Los resultados sugieren que durante la reperfusión se produce un daño miocárdico que en parte es causado por la citotoxicidad de los radicales libres del oxígeno. Se concluye además que la deferoxamina y más el manitol reducen las lesiones de reperfusión y que su mecanismo de acción sería por la capacidad antioxidante que poseen ambas sustancias