Purple pitanga extract (Eugenia uniflora) attenuates oxidative stress induced by MPTP.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) has been widely used due to its specific and reproducible neurotoxic effect on the nigrostriatal system, being considered a convenient model of dopaminergic neurodegeneration to study interventions therapeutics. The purple pitanga (Eugenia uniflora) is a polyphenol-rich fruit with antioxidant and antidepressant properties, among others. Therefore, this study investigated the effect of purple pitanga extract (PPE) on acute early oxidative stress induced by intranasal 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration in rats. Male Wistar rats were pre-treated orally with PPE (1000 mg/kg) or vehicle. After 24 h, MPTP (0.1 mg/10µL/nostril) or vehicle was administered bilaterally into the animal's nostrils, and 6 h later, the olfactory bulb (OB), striatum (ST), and substantia nigra (SN) were collected to evaluate the oxidative stress parameters. Our findings revealed that OB and SN were the most affected areas after 6 h of MPTP infusion; an early increase in reactive oxygen species (ROS) levels was observed, while pretreatment with a single dose of PPE prevented this increment. No differences in thiobarbituric acid reactive species (TBARS) and 3-nitrotyrosine (3-NT) formation were observed, although 4-hydroxy-2-nonenal (4-HNE) levels increased, which is the most toxic form of lipid peroxidation, in the MPTP group. The PPE pretreatment could prevent this increase by increasing the NPSH levels previously decreased by MPTP. Furthermore, PPE prevents the Na+/K + ATPase strongly inhibited by MPTP, showing the neuroprotective capacity of the PPE by inhibiting the MPTP-generated oxidation. Thus, we demonstrated for the first time the antioxidant and neuroprotective effects of PPE against the early MPTP neurotoxicity.

Neuroprotective effect of Eugenia uniflora against intranasal MPTP-induced memory impairments in rats: The involvement of pro-BDNF/p75NTR pathway.

Parkinson's disease is a multisystemic neurodegenerative disorder that includes motor and non-motor symptoms, and common symptoms include memory loss and learning difficulties. Thus, we investigated the neuroprotective potential of a hydroalcoholic extract of Brazilian purple cherry (Eugenia uniflora) (HAE-BC) on memory impairments induced by intranasal 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration in rats and the involvement of hippocampal BDNF/TrkB/p75NTR pathway in its effects. Adult male Wistar rats were exposed to MPTP (1 mg/nostril) or vehicle. Twenty-four hours later, the HAE-BC treatments began at doses of 300 or 2000 mg/kg/day or vehicle for 14 days. From 7 days after the MPTP induction, the animals were subjected to behavioral tests to evaluate several cognitive paradigms. HAE-BC treatments, at both doses, blocked the MPTP-caused disruption in the social recognition memory, short- and long-term object recognition memories, and working memory. Furthermore, MPTP-induced motor deficit linked to striatal tyrosine hydroxylase levels decreased, which was blocked by HAE-BC. Our findings demonstrated that HAE-BC blocked the MPTP-induced increase in the hippocampal pro-BDNF, TrkB.t1, and p75NTR levels. The pro-BDNF/p75NTR interaction negatively regulates synaptic transmission and plasticity, and the neuroprotective effect of HAE-BC was related, at least partly, to the modulation of this hippocampal signaling pathway. Thus, our study reports the first evidence of the potential therapeutic of E. uniflora in a Parkinson's disease model in rodents.

Role of the antioxidant pathway in the healing of peptic ulcers induced by ischemia-reperfusion in male and female rats treated with Eugenia punicifolia.

Ischaemia and reperfusion (I/R)-induced gastrointestinal disorders are caused by free radicals, resulting in organ damage and functional disarrangement. This study aimed to investigate the healing effects of hydroalcoholic extracts from the leaves of Eugenia punicifolia (Kunth) DC. (HEEP) in male and female Wistar rats with I/R-induced peptic injuries, and the role of antioxidants in improving this response. After I/R-induced gastric and duodenal injuries, male and female [intact (INT) and ovariectomized (OVZ)] rats were orally treated with HEEP for 6 days. Biochemical analysis was used to determine the catalase (CAT), superoxide dismutase (SOD), and myeloperoxidase (MPO) activities, as well as malondialdehyde and reduced glutathione levels, to measure the gastric and duodenal healing process. Six days of HEEP treatment significantly decreased the I/R-induced gastric [male (73.68%), INT (52.83%), and OVZ (43.13%)] and duodenal damage [male (57.03%), INT (56.04%), and OVZ (54.83%)] in all groups. In OVZ rats, the healing effect of HEEP occurred because of the increased activity of SOD (2x) and CAT (1.16x) in the gastric mucosa. In the duodenal mucosa of INT rats, the extract reduced MPO (20.83%) activity. The 6-day HEEP treatment improved the healing of I/R-induced peptic ulcer injury, with the system acting differently in males and females. The antioxidant system is an important component of the HEEP activity during post-I/R mucosal recovery. This result revealed the importance of antioxidant compounds in minimizing the severity of I/R-related events.

Chemical Composition and Antioxidant Activity of Essential Oils from Eugenia patrisii Vahl, E. punicifolia (Kunth) DC., and Myrcia tomentosa (Aubl.) DC., Leaf of Family Myrtaceae.

Essential oils (EOs) were extracted from Eugenia patrisii, E. punicifolia, and Myrcia tomentosa, specimens A and B, using hydrodistillation. Gas chromatography coupled with mass spectrometry (GC/MS) was used to identify the volatile constituents present, and the antioxidant capacity of EOs was determined using diphenylpicryl-hydrazyl (DPPH) and trolox equivalent antioxidant capacity (TEAC) assays. For E. patrisii, germacrene D (20.03%), bicyclogermacrene (11.82%), and (E)-caryophyllene (11.04%) were identified as the major constituents of the EOs extracted from specimen A, whereas specimen B primarily comprised γ-elemene (25.89%), germacrene B (8.11%), and (E)-caryophyllene (10.76%). The EOs of E. punicifolia specimen A contained ß-Elemene (25.12%), (E)-caryophyllene (13.11%), and bicyclogermacrene (9.88%), while specimen B was composed of (E)-caryophyllene (11.47%), bicyclogermacrene (5.86%), ß-pinene (5.86%), and γ-muurolene (5.55%). The specimen A of M. tomentosa was characterized by γ-elemene (12.52%), germacrene D (11.45%), and (E)-caryophyllene (10.22%), while specimen B contained spathulenol (40.70%), α-zingiberene (9.58%), and γ-elemene (6.89%). Additionally, the chemical composition of the EOs was qualitatively and quantitatively affected by the collection period. Furthermore, the EOs of the studied specimens, especially specimen A of E. punicifolia, showed a greater antioxidant activity in DPPH rather than TEAC, as represented by a significantly high inhibition percentage (408.0%).

Physicochemical Properties and Effects of Fruit Pulps from the Amazon Biome on Physiological Parameters in Rats.

This study aimed to analyze the physicochemical characteristics and the effects of Amazonian pulp fruits consumption, such as araçá-boi (Eugenia stipitata), abiu grande (Pouteria caimito), araticum (Annona crassiflora), biri-biri (Averrhoa bilimbi L.), and yellow mangosteen (Garcinia xanthochymus), on hematologic, metabolic, renal, and hepatic function parameters in Wistar rats (n = 10 rats/group). The pulp of abiu had the highest levels of soluble solids, sugars, and pH. Biri-biri pulp had the highest levels of ascorbic acid and total titratable acidity, and a low pH. The araticum pulp had higher (p ≤ 0.05) ash content, total phenolic compounds, and antioxidant activity than the pulp of other analyzed fruits. No significant increase in hematocrit, nor reduction of blood glucose, plasma cholesterol, and serum levels of glutamic-pyruvic transaminase (TGP), creatinine, and urea was observed in experimental groups relative to the control group of rats after the consumption of fruits pulp. The intake of abiu and araticum pulps promoted a significant reduction (p ≤ 0.05) in total leukocytes of the experimental groups as compared to the control group and only the intake of araticum significantly increased (p ≤ 0.05) triglyceride blood levels in rats (99.50 mg/dL). The regular consumption of biri-biri pulp for 30 days significantly (p ≤ 0.05) increased serum glutamic-oxaloacetic transaminase (TGO) levels in rats (116.83 U/L) compared to the control group (98.00 U/L). More researches are needed to generate knowledge about these promising Amazonian fruits, supporting the native fruit production, in addition to promoting health in the population and sustainability in the Amazon region.

Influence of high-intensity ultrasound on color, chemical composition and antioxidant properties of araçá-boi pulp.

In this study, we evaluated the influence of the ultrasound application on five levels of energy density (1000; 3000; 5000 and 7000 J g-1) compared to two pasteurization techniques (70 °C/5 min and 94 °C/0.5 min) on color parameters, polyphenoloxidase activity, chemical composition, and antioxidant properties of araçá-boi pulp. Ultrasound caused changes in the parameters brightness/darkness, hue angle, and total color difference, but did not change chroma, yellowness/blueness, color index, and yellow index. Moreover, this technique was efficient for inactivating polyphenoloxidase. Ultrasound at 7000 J g-1 was responsible for an increase in soluble solids (16%), vitamin C (46.5%), phenolics (15.65%), flavonoids (50%) and antioxidant capacity in relation to untreated pulp, while ultrasound at 5000 J g-1 increased the relative intensity of compounds of biological interest. Thus, ultrasound can be considered as a promising technique to maintain the shelf life, without drastically affecting the nutritional and functional qualities of this fruit.

Characterization and expression analysis of P5CS (Δ1-pyrroline-5-carboxylate synthase) gene in two distinct populations of the Atlantic Forest native species Eugenia uniflora L.

Eugenia uniflora is an Atlantic Forest native species, occurring in contrasting edaphoclimatic environments. The identification of genes involved in response to abiotic factors is very relevant to help in understanding the processes of local adaptation. 1-Pyrroline-5-carboxylate synthetase (P5CS) is one interesting gene to study in this species since it encodes a key enzyme of proline biosynthesis, which is an osmoprotectant during abiotic stress. Applying in silico analysis, we identified one P5CS gene sequence of E. uniflora (EuniP5CS). Phylogenetic analysis, as well as, gene and protein structure investigation, revealed that EuniP5CS is a member of P5CS gene family. Plants of E. uniflora from two distinct environments (restinga and riparian forest) presented differences in the proline accumulation and P5CS expression levels under growth-controlled conditions. Both proline accumulation and gene expression level of EuniP5CS were higher in the genotypes from riparian forest than those from restinga. When these plants were submitted to drought stress, EuniP5CS gene was up-regulated in the plants from restinga, but not in those from riparian forest. These results demonstrated that EuniP5CS is involved in proline biosynthesis in this species and suggest that P5CS gene may be an interesting candidate gene in future studies to understand the processes of local adaptation in E. uniflora.

Effect of oregano (Origanum vulgare L. ssp. hirtum) and clove (Eugenia spp.) nanoemulsions on Zygosaccharomyces bailii survival in salad dressings.

This work aimed to evaluate the in vitro effect of encapsulated oregano and clove essential oils on oil-in-water nanoemulsions against Zygosaccharomyces bailii. The antifungal efficacy of these nanoemulsions and their sensory acceptance were tested in salad dressings. Both essential oils were effective inhibitors against the target yeast, with minimal inhibitory and fungicidal concentrations of 1.75 mg/mL. In the in vitro assay done with the nanoemulsions, no yeast growth was observed for any tested essential oil concentration. In the salad dressings, all the formulations were able to reduce Z. bailii growth compared to the control, and only those samples with 1.95 mg/g of essential oil were capable of inhibiting yeast development after 4 inoculation days. The sensory acceptance of the dressing containing the nanoemulsions was similar to the control dressing in appearance, consistency and colour terms. These results evidence the antifungal activity of oregano and clove nanoemulsions against Z. bailii.

Chemical profiling of secondary metabolites of Eugenia uniflora and their antioxidant, anti-inflammatory, pain killing and anti-diabetic activities: A comprehensive approach.

ETHNOPHARMACOLOGICAL RELEVANCE: The red Brazilian cherry, Eugenia uniflora, is widely used in traditional medicine. The aim of this study was to investigate the phytochemical composition of a methanol extract from leaves of E. uniflora and characterization of the isolated compounds. In addition, we aimed to determine the antioxidant activities in vitro and in a cell-based (HaCaT cell) model. We also studied the anti-inflammatory, analgesic, antipyretic and antidiabetic activities in relevant rat models. The molecular mode of action of the antidiabetic activities was also investigated. MATERIALS AND METHODS: UV, MS, and NMR (1H, 13C, DEPT, COSY, HMQC, and HMBC) were used to identify the secondary metabolites. Antioxidant effects were determined in vitro and in HaCaT cells. The ani-inflammatory and antidibetic activities were studied in experimental animals. RESULTS: In this work, a new compound, gallic acid 3-O-[6'-O-acetyl-ß-D-glucoside], along with 16 known plant secondary metabolites (PSM) were isolated, characterized using UV, MS, and NMR (1H, 13C, DEPT, COSY, HMQC, and HMBC). Noticeable antioxidant effects were determined in HaCaT cells: The extract reduced the elevated levels of ROS and p38 phosphorylation and increased the reduced glutathione (GSH) content induced by UVA. The extract showed substantial anti-inflammatory activities in vivo: It diminished the edema thickness in carrageenan-induced hind-paw edema rat model and lowered the leukocyte migration into the peritoneal cavity. In rats, central and peripheral anti-nociceptive properties were also observed: The extract reduced the number of writhing in acid induced writhing and increased the latency time in hot plate test. Furthermore, adequate antipyretic effects were observed: The extract reduced the elevated rectal temperature in rats after intraperitoneal injection of Brewer's yeast. Moreover, the extract possessed robust anti-diabetic activity in streptozotocin (STZ) -diabetic rats: It markedly reduced the elevated serum glucose and lipid peroxidation levels and increased the insulin concentration in serum with higher potency than the positive control, glibenclamide. These effects might be associated with the interaction of PSM with the conserved amino acid residues of human pancreatic α-amylase (HPA), maltase glucoamylase (MGAM-C) and aldose reductase (ALR2) revealing considerable binding affinities. CONCLUSION: A plethora of substantial pharmacological properties indicates that Eugenia uniflora is a good antioxidant and a sustainable by-product with solid therapeutic potential for treating diabetes, inflammation, pain and related oxidative stress diseases.

Chemical composition, nutritional value and bioactive compounds in six uvaia accessions.

This study analyzed six uvaia (Eugenia pyriformis Cambess) accessions ('Comum', 'Rugosa', 'Doce de Patos de Minas', 'Pêra', 'Rugosa Doce' and 'Dura') with respect to chemical composition, nutritional value, bioactive compounds and antioxidant capacity. The 'Rugosa Doce' and 'Doce de Patos de Minas' accessions were sweeter (20.18 g 100 g-1 and 18.88 g 100 g-1, respectively), while the 'Rugosa' and 'Pêra' accessions were less acid (0.28 g 100 g-1 and 0.33 g 100 g-1, respectively). Terpenes were the major volatile compounds (46.75%) identified by SPME-GC/MS. The 'Rugosa' accession contained high levels of fibers (43.81 g 100 g-1) and minerals (2235.10  mg 100 g-1). The 'Rugosa Doce' and 'Rugosa' accessions were distinguished by total flavonoids and phenolic compounds. 'Rugosa Doce' and 'Rugosa' may potentially serve as a raw material for food, while 'Doce de Patos de Minas' and 'Pêra' can be used for cosmetic and pharmaceutical industries purposes.

Chlorogenic and caffeic acids in 64 fruits consumed in Brazil.

This study presents unpublished data on the chlorogenic acid and caffeic acid content in fruits. Sixty-four fruits consumed in Brazil, most of which were produced domestically, were evaluated based on their levels of 3-caffeoylquinic acid, 4-caffeoylquinic acid, 5-caffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and caffeic acid. The study investigated 15 fruits that have not been reported in the literature in relation to these compounds, including several native species. The highest concentration of mono-caffeoylquinic acid was observed in strawberry, cherry, bilberry, quince and mulberry, while the dicaffeoylquinic acid was present with highest concentration in kumquat, passion fruit and sweet granadilla. Regarding caffeic acid, the highest content was found in bilberry and yellow pitaya. Considering the sum of the concentrations of these compounds, quince, cherry, bilberry, mulberry and sweet granadilla were exceptional, with concentrations between 200.0 and 569.7 mg kg-1.

Important issues in plant tissues analyses by HR-MAS NMR.

INTRODUCTION: High-resolution magic-angle spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy enables the analysis of the metabolic profile of plant and animal tissues under close to natural conditions, as well as of other heterogeneous natural or synthetic materials. Neither sample pretreatment is required after fragmentation nor powdering of the sample before insertion into the rotor. However, the efficiency of the method depends strongly on the sample preparation, rotor insertion procedure, and analysis conditions. OBJECTIVE: To identify some of the variables that affect the spectral data and to propose solutions that minimise their impact on the quality of the analyses and results. METHODS: Dried plant tissues were powdered, weighed, and homogenised in a 50 µL rotor with an optimised volume of deuterated solvent and sample in order to prevent material from escaping during spacer insertion, avoiding variations in magnetic susceptibility. Factors affecting the quality of HR-MAS NMR analysis such as particle size, sample and solvent amounts, solvent polarity, swelling time, rotor manipulation and pulse sequence setting were evaluated. RESULTS: A strong correlation was observed between the signal area and the particle size of the powdered sample. The spectral profile varied depending on the deuterated solvent used. An incubation period was necessary to achieve adequate swelling of the sample and to ensure good data reproducibility. Proper sealing of the rotor, number of cycles and τ time on cpmgpr1d pulse sequence were found to affect the signal areas. CONCLUSION: The study highlights the need for standardised sample preparation and instrumental setup protocols in order to achieve high reproducibility and obtain reliable data from HR-MAS NMR analyses.

Effect of in vitro digestion on the functional properties of Psidium cattleianum Sabine (araçá), Butia odorata (Barb. Rodr.) Noblick (butiá) and Eugenia uniflora L. (pitanga) fruit extracts.

Brazilian native fruits are reported to be promising sources of bioactive compounds; however their bioactivity depends on their stability along the digestive process. This study evaluated the α-glucosidase inhibition, antioxidant activity and total phenolic content (TPC) stability of araçá, butiá and pitanga fruit extracts using an in vitro digestion model. Additionally, the individual phenolic compound recovery of the most stable and active extract was evaluated by HPLC-DAD-ESI-MS/MS. Overall, the antioxidant activity of all extracts decreased along the process. Araçá fruit extracts, at the end of digestion, showed α-glucosidase inhibition values similar to their non-digested extracts and the highest TPC recovery (28%). Recovery of individual phenolic compounds of red araçá fruit extract revealed a negative impact on the stability of ellagitannins. Araçá fruit extract seems to provide phenolic compounds with α-glucosidase inhibitory properties after the gastrointestinal digestion, indicating their potential to be used in the control of type II diabetes.

Clusia hilariana and Eugenia uniflora as bioindicators of atmospheric pollutants emitted by an iron pelletizing factory in Brazil.

The objectives of this work were to evaluate if the pollution emitted by the pelletizing factory causes visual symptoms and/or anatomical changes in exposed Eugenia uniflora and Clusia hilariana, in active biomonitoring, at different distances from a pelletizing factory. We characterize the symptomatology, anatomical, and histochemistry alterations induced in the two species. There was no difference in the symptomatology in relation to the different distances of the emitting source. The foliar symptoms found in C. hilariana were chlorosis, necrosis, and foliar abscission and, in E. uniflora, were observed necrosis punctuais, purple spots in the leaves, and increase in the emission of new leaves completely purplish. The two species presented formation of a cicatrization tissue. E. uniflora presented reduction in the thickness of leaf. In C. hilariana, it was visualized hyperplasia of the cells and the adaxial epidermis did not appear collapsed due to thick cuticle and cuticular flanges. Leaves of C. hilariana showed positive staining for iron, protein, starch, and phenolic compounds. E. uniflora showed positive staining for total phenolic compounds and starch. Micromorphologically, there was accumulation of particulate matter on the leaf surface, obstruction of the stomata, and scaling of the epicuticular wax in both species. It was concluded that the visual and anatomical symptoms were efficient in the diagnosis of the stress factor. C. hilariana and E. uniflora showed to be good bioindicators of the atmospheric pollutants emitted by the pelletizing factory.

Potential antiproliferative activity of polyphenol metabolites against human breast cancer cells and their urine excretion pattern in healthy subjects following acute intake of a polyphenol-rich juice of grumixama (Eugenia brasiliensis Lam.).

The bioavailability and metabolism of anthocyanins and ellagitannins following acute intake of grumixama fruit, native Brazilian cherry, by humans, and its in vitro antiproliferative activity against breast cancer cells (MDA-MB-231) were investigated. A single dose of grumixama juice was administered to healthy women (n = 10) and polyphenol metabolites were analyzed in urine and plasma samples collected over 24 h. The majority of the metabolites circulating and excreted in urine were phenolic acids and urolithin conjugates, the gut microbiota catabolites of both classes of polyphenols, respectively. According to pharmacokinetic parameters, the subjects were divided into two distinct groups, high and low urinary metabolite excretors. The pool of polyphenol metabolites found in urine samples showed a significant inhibition of cell proliferation and G2/M cell cycle arrest in MDA-MB-231 cells. Our findings demonstrate the large interindividual variability concerning the polyphenol metabolism, which possibly could reflect in health promotion.

Physiological and biochemical mechanisms of the ornamental Eugenia myrtifolia L. plants for coping with NaCl stress and recovery.

MAIN CONCLUSION: We studied the response of Eugenia myrtifolia L. plants, an ornamental shrub native to tropical and subtropical areas, to salt stress in order to facilitate the use of these plants in Mediterranean areas for landscaping. E. myrtifolia plants implement a series of adaptations to acclimate to salinity, including morphological, physiological and biochemical changes. Furthermore, the post-recovery period seems to be detected by Eugenia plants as a new stress situation. Different physiological and biochemical changes in Eugenia myrtifolia L. plants after being subjected to NaCl stress for up to 30 days (Phase I) and after recovery from salinity (Phase II) were studied. Eugenia plants proved to be tolerant to NaCl concentrations between 44 and 88 mM, displaying a series of adaptative mechanisms to cope with salt-stress, including the accumulation of toxic ions in roots. Plants increased their root/shoot ratio and decreased their leaf area, leaf water potential and stomatal conductance in order to limit water loss. In addition, they displayed different strategies to protect the photosynthetic machinery, including the limited accumulation of toxic ions in leaves, increase in chlorophyll content, changes in chlorophyll fluorescence parameters, leaf anatomy and antioxidant defence mechanisms. Anatomical modifications in leaves, including an increase in palisade parenchyma and intercellular spaces and decrease in spongy parenchyma, served to facilitate CO2 diffusion in a situation of reduced stomatal aperture. Salinity produced oxidative stress in Eugenia plants as evidenced by oxidative stress parameters values and a reduction in APX and ASC levels. Nevertheless, SOD and GSH contents increased. The post-recovery period is detected as a new stress situation, as observed through effects on plant growth and alterations in chlorophyll fluorescence and oxidative stress parameters.