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In this study, we investigated the influence of mixture design on the chemical profile of Eugenia unifloraleaves, evaluating the antioxidant and antimicrobial activities, the toxic and hemolytic potential, with the focus on the improvement of the polyphenol's extraction for incorporation of the extract in semi-solid forms with antifungal action. The chemical analysis was evaluated by UV-Vis and HPLC. The antimicrobial, antioxidant, and hemolytic activities were monitored. The flavonoid content ranged from 2.63-7.98 %w/w and tannins from 5.42-18.29 %w/w. The extract consisted of gallic acid (0.09-1.29%; w/w), ellagic acid (0.09-0.37%; w/w), and myricitrin (0.18-1.20%; w/w). The most successful solvent system with the highest level of active extract was water: ethanol: propylene glycol. The extracts showed fungicidal properties (3.9 µg/mL), high antioxidant activity (IC50: 9.50 µg/mL), and low toxicity. These solvent mixtures can improve the in vitro bioactivities when compared to pure solvents and this result demonstrates the importance of mixture designs as useful tools for creating high-quality herbal products and elucidate the potential of E. uniflora glycolic extracts as active herbal pharmaceutical ingredients in topical delivery systems.

En este estudio investigamos la influencia del diseño de mezclas en el perfil químico de hojas de Eugenia uniflora, evaluando las actividades antioxidantes y antimicrobianas, el potencial tóxico y hemolítico, con el foco en la mejora de la extracción de polifenoles para la incorporación del extracto en formas semi-sólidas con acción antifúngica. El análisis químico se evaluó mediante UV-Vis y HPLC. Se monitorizaron las actividades antimicrobianas, antioxidantes y hemolíticas. El contenido de flavonoides osciló entre 2,63 y 7,98% p/p and taninos de 5,42-18,29% p/p. El extracto consistió en ácido gálico (0.09-1.29%; p/p), ácido elágico (0.09-0.37%; p/p) y miricitrina (0.18-1.20%; p/p). El sistema de disolventes más exitoso con el nivel más alto de extracto activo fue agua: etanol: propilenglicol. Los extractos mostraron propiedades fungicidas (3.9 µg/mL), alta actividad antioxidante (IC50: 9.50 µg/mL) y baja toxicidad. Estas mezclas de disolventes pueden mejorar las bioactividades in vitro en comparación con los disolventes puros y este resultado demuestra la importancia de los diseños de mezclas como herramientas útiles para crear productos a base de hierbas de alta calidad y dilucidar el potencial de los extractos glicólicos de E. uniflora como ingredientes farmacéuticos a base de hierbas en sistemas de entrega activos.

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BACKGROUND: Fetal stage is a critical developmental window for the skeletal muscle, but little information is available about the impact of maternal vitamin D (Vit. D) deficiency (VDD) on offspring lean mass development in the adult life of male and female animals. METHODS: Female rats (Wistar Hannover) were fed either a control (1000 IU Vit. D3/kg) or a VDD diet (0 IU Vit. D3/kg) for 6 weeks and during gestation and lactation. At weaning, male and female offspring were randomly separated and received a standard diet up to 180 days old. RESULTS: Vitamin D deficiency induced muscle atrophy in the male (M-VDD) offspring at the end of weaning, an effect that was reverted along the time. Following 180 days, fast-twitch skeletal muscles [extensor digitorum longus (EDL)] from the M-VDD showed a decrease (20%; P < 0.05) in the number of total fibres but an increase in the cross-sectional area of IIB (17%; P < 0.05), IIA (19%; P < 0.05) and IIAX (21%; P < 0.05) fibres. The fibre hypertrophy was associated with the higher protein levels of MyoD (73%; P < 0.05) and myogenin (55% %; P < 0.05) and in the number of satellite cells (128.8 ± 14 vs. 91 ± 7.6 nuclei Pax7 + in the M-CTRL; P < 0.05). M-VDD increased time to fatigue during ex vivo contractions of EDL muscles and showed an increase in the phosphorylation levels of IGF-1/insulin receptor and their downstream targets related to anabolic processes and myogenic activation, including Ser 473 Akt and Ser 21/9 GSK-3ß. In such muscles, maternal VDD induced a compensatory increase in the content of calcitriol (two-fold; P < 0.05) and CYP27B1 (58%; P < 0.05), a metabolizing enzyme that converts calcidiol to calcitriol. Interestingly, most morphological and biochemical changes found in EDL were not observed in slow-twitch skeletal muscles (soleus) from the M-VDD group as well as in both EDL and soleus muscles from the female offspring. CONCLUSIONS: These data show that maternal VDD selectively affects the development of type-II muscle fibres in male offspring rats but not in female offspring rats and suggest that the enhancement of their size and fatigue resistance in fast-twitch skeletal muscle (EDL) is probably due to a compensatory increase in the muscle content of Vit. D in the adult age.

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Alterations in the renal vasculature during fetal programming can cause disturbances in renal structure and function that persist into adulthood. Calcitriol can affect cellular differentiation and proliferation, and promote endothelial cell maintenance, each of which is a key event in nephrogenesis. Calcitriol is a negative endocrine regulator of the renin gene. Rats exposed to renin-angiotensin system (RAS) antagonists during lactation have been shown to develop renal disorders, which demonstrated that the RAS may play an important role in mammalian kidney development. We evaluated the effects of calcitriol administration on losartan [angiotensin II receptor antagonist (ANGII), AT1]-induced changes in renal differentiation in rats during lactation. Rats treated with losartan showed alterations in renal function and structure that persisted into adulthood. These disruptions included hydronephrosis, papillary atrophy, endothelial dysfunction, and aberrant endothelial structure. These changes were mitigated by treatment with calcitriol. The results of our study showed that animals exposed to AT1 blockade during lactation exhibited altered renal microvasculature differentiation in adulthood that was attenuated by treatment with calcitriol.

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BACKGROUND/AIMS: Physical training has beneficial effects on endothelial function and can influence the regeneration of the endothelial cell. We investigated the effect of physical training on cisplatin (CP)-induced acute kidney injury and assessed the impact of training on endothelial structure and function, and on the inflammatory processes in rats. METHODS: We injected male Wistar rats subjected to previous physical training in treadmill running (trained, TR) or not (sedentary, SED) with CP (5 mg/kg) (TR+CP and SED+CP groups, respectively). Five days after the injections, blood and urine samples were collected to evaluate renal function and kidneys were harvested for morphological, immunohistochemical, enzyme-linked immunosorbent assay, and analysis of nitric oxide (NO) levels. RESULTS: Rats treated with CP showed increased levels of plasma creatinine and sodium and potassium fractional excretion. These alterations were associated with increase in tubulointerstitial lesions and macrophage number, reduction of endothelial cells, and increased VEGF, vimentin, and α-smooth muscle actin expression in the outer renal medulla in the SED+CP group. We also found increased levels of renal IL-1ß and increased excretion of monocyte chemoattractant protein-1 and transforming growth factor-ß compared with controls. These changes were milder in trained rats, associated with increased levels of renal tissue NO, and increased expression of p-eNOS and stromal cell-derived factor-1α (a chemokine involved in kidney repair) in the kidneys of CP-injected trained rats. CONCLUSIONS: The protective effect of previous training in CP-treated rats was associated with reduced endothelial cell lesions and increased renal production of NO in trained rats.

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Excessive sodium (Na+) intake in modern society has been associated with several chronic disorders such as hypertension. Several studies suggest that early life events can program physiological systems and lead to functional changes in adulthood. Therefore, we investigated behavioral and neuroendocrine responses under basal conditions and after 48 h of water deprivation in adult (60-day-old Wistar rats) male, Wistar rats originating from dams were offered only water or 0.15 mol/L NaCl during pregnancy and lactation. Early life salt exposure induced kidney damage, as shown by a higher number of ED-1 positive cells (macrophages/monocytes), increased daily urinary volume and Na+ excretion, blunted basal water intake and plasma oxytocin levels, and increased plasma corticosterone secretion. When challenged with water deprivation, animals exposed to 0.15 mol/L NaCl during early life showed impaired water intake, reduced salt preference ratio, and vasopressin (AVP) secretion. In summary, our data demonstrate that the perinatal exposure to excessive Na+ intake can induce kidney injury in adult offspring and significantly affect the key mechanisms regulating water balance, fluid intake, and AVP release in response to water deprivation. Collectively, these novel results highlight the impact of perinatal programming on the homeostatic mechanisms regulating fluid and electrolyte balance during exposure to an environmental stress (i.e. dehydration) in later life.

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