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    Denise Endringer

    Resumo A fitoterapia popular se baseia no conhecimento tradicional de plantas medicinais, muitas vezes sem comprovação de suas propriedades terapêuticas. Diversos estudos têm demonstrado que fitoestrogênios seriam alternativas naturais à... more
    Resumo A fitoterapia popular se baseia no conhecimento tradicional de plantas medicinais, muitas vezes sem comprovação de suas propriedades terapêuticas. Diversos estudos têm demonstrado que fitoestrogênios seriam alternativas naturais à terapia de reposição hormonal (TRH). O objetivo do estudo foi avaliar o efeito do "hormônio natural", almeirão-roxo, na TRH em ratas com menopausa cirúrgica. Estas foram divididas em três grupos:
    Increased tea consumption in combination with intensive pesticide use is generating heavy metal contaminations amongst Brazilian tea consumers, causing health concerns. Inductively coupled plasma optical emission spectrometry (ICP-OES)... more
    Increased tea consumption in combination with intensive pesticide use is generating heavy metal contaminations amongst Brazilian tea consumers, causing health concerns. Inductively coupled plasma optical emission spectrometry (ICP-OES) was applied to quantify minerals and heavy metals such as aluminum, barium, cadmium, lead, cobalt, copper, chromium, tin, manganese, molybdenum, nickel, selenium, silver, thallium, vanadium and zinc in Brazilian chamomile, lemongrass, fennel and yerba mate teas. Teas, purchased in local supermarkets, were prepared using infusion and acid digestion. Higher concentrations of Al were present in all samples. In the digested samples, the Al mean concentration was 2.41 μg g(-1) (sd = 0.72) for fennel and 33.42 μg g(-1) (sd = 17.18) for chamomile, whilst the sample C for chamomile tea presented the highest concentration with 51.62 μg g(-1) (sd = 9.17). The safety relation in decreasing order is fennel, lemongrass, chamomile and yerba mate. Chemometric analys...
    Hedychium coronarium J. Koenig, Zingiberaceae, is a medicinal plant popularly used to treat inflammatory conditions in different countries. Three labdane diterpenes [isocoronarin D (1), methoxycoronarin D (2), ethoxycoronarin D (3)] and... more
    Hedychium coronarium J. Koenig, Zingiberaceae, is a medicinal plant popularly used to treat inflammatory conditions in different countries. Three labdane diterpenes [isocoronarin D (1), methoxycoronarin D (2), ethoxycoronarin D (3)] and benzoyl eugenol (4) were isolated from rhizomes and their chemopreventive potential was evaluated using in vitro assays, namely the inhibition of NF-κB, COX-1 and -2, the induction of antioxidant response element (ARE), and the inhibition of cell proliferation. Diterpene 1 activated ARE (EC50 57.6 ± 2.4 μM), while 2, 3 and 4 significantly inhibited NF-κB (IC50 of 7.3 ± 0.3, 3.2 ± 0.3 and 32.5 ± 4.9 μM, respectively). In addition, 2 and 3 selectively inhibited COX-1 (IC50 values of 0.9 ± 0.0 and 3.8 ± 0.0 μM, respectively). These data support the potential chemopreventive activity of constituents from H. coronarium rhizomes.
    The negative relationship between androgens and the Bezold-Jarisch reflex (BJR) has been demonstrated, but no studies evaluated the physiological influence of testosterone on this reflex. We evaluated the influence of male rat castration... more
    The negative relationship between androgens and the Bezold-Jarisch reflex (BJR) has been demonstrated, but no studies evaluated the physiological influence of testosterone on this reflex. We evaluated the influence of male rat castration on the BJR, cardiac morphometric parameters, and the plasmatic and the cardiac angiotensin converting enzyme (ACE) activity. After castration (CAS), the rats were divided into 24 and 72 h (CAS24H, CAS72H), and 7 and 21 days (CAS7D, CAS21D) groups. The BJR was studied by administering increasing doses of phenylbiguanide (PBG; 1.5-24 μg/kg) at different times after castration. Castration results in the following: (i) reduction in testosterone levels (SHAM: 238.7 ± 15.1; CAS24H: 9.0 ± 0.5; CAS72H: 6.7 ± 0.4; CAS7D: 5.2 ± 0.2; and CAS21D: 2.2 ± 0.3 ng/dL; p < 0.05); (ii) no changes in 17β-estradiol; (iii) a reduced BJR sensitivity (PBG 6 μg/kg; SHAM: 77 ± 7; CAS24H: 63 ± 10; CAS72H: 55 ± 6; CAS7D: 54 ± 4; and CAS21D: 35 ± 2%; p < 0.01); (iv) a decrease in cardiac (SHAM: 107 ± 6; CAS24H: 92 ± 2; CAS72H: 82 ± 3; CAS7D: 54 ± 3; and CAS21D: 43 ± 4%; p < 0.05) and plasmatic (SHAM: 135 ± 8; CAS24H: 102 ± 5; CAS72H: 99 ± 3; CAS7D: 89 ± 4; and CAS21D: 56 ± 6%; p < 0.05) ACE activity. No changes were observed in cardiac morphometry and hemodynamic parameters. Therefore, castration leads to decrease in testosterone levels as early as 24 h, reduction in ACE activity and loss of BJR sensitivity 7 days after castration. The loss of BJR sensitivity was not related to cardiac morphometric changes and cardiovascular hemodynamics.
    ABSTRACT Background: Cellular viability studies are important in many different fields of cell biology research and are based on estimates of live and dead cells from a specific cellular population. Recent progress in technology has... more
    ABSTRACT Background: Cellular viability studies are important in many different fields of cell biology research and are based on estimates of live and dead cells from a specific cellular population. Recent progress in technology has brought important advances in the analysis of cellular viability, mainly reducing the time of analysis and sample preparation; however, the costs for equipment and reagents still limit the feasibility of conducting those studies. The purpose of the present study was to use free software, non-specific dyes, and light microscopy to establish a reproducible image analysis test and to compare the results with an established method. Method: 3T3 fibroblast cell line was exposed to different concentrations of dimethylsulfoxide (DMSO) for 24 h, and cellular viability was estimated by the proposed technique (image analysis performed with free software) and with the methylthiazol-tetrazolium salt (MTT) assay. A Bland-Altman analysis was applied to compare the methodologies. Results: Image analysis by CellProfiler®/CellProfiler® Analyst was compared to MTT by Bland-Altman test, and the two methodologies were considered equivalent. Cellular viability decreased in a dose-dependent manner with increasing doses of DMSO, and both methodologies were capable of distinguishing between live and dead cells, producing comparable results (with bias = 1.3550) evaluated by the Bland-Altman analysis. Conclusion: The proposed image analysis can be used as a simple, rapid, and low-cost technology for high throughput analysis of live-dead cell differentiation.
    ABSTRACT Background: Cellular viability studies are important in many different fields of cell biology research and are based on estimates of live and dead cells from a specific cellular population. Recent progress in technology has... more
    ABSTRACT Background: Cellular viability studies are important in many different fields of cell biology research and are based on estimates of live and dead cells from a specific cellular population. Recent progress in technology has brought important advances in the analysis of cellular viability, mainly reducing the time of analysis and sample preparation; however, the costs for equipment and reagents still limit the feasibility of conducting those studies. The purpose of the present study was to use free software, non-specific dyes, and light microscopy to establish a reproducible image analysis test and to compare the results with an established method. Method: 3T3 fibroblast cell line was exposed to different concentrations of dimethylsulfoxide (DMSO) for 24 h, and cellular viability was estimated by the proposed technique (image analysis performed with free software) and with the methylthiazol-tetrazolium salt (MTT) assay. A Bland-Altman analysis was applied to compare the methodologies. Results: Image analysis by CellProfiler®/CellProfiler® Analyst was compared to MTT by Bland-Altman test, and the two methodologies were considered equivalent. Cellular viability decreased in a dose-dependent manner with increasing doses of DMSO, and both methodologies were capable of distinguishing between live and dead cells, producing comparable results (with bias = 1.3550) evaluated by the Bland-Altman analysis. Conclusion: The proposed image analysis can be used as a simple, rapid, and low-cost technology for high throughput analysis of live-dead cell differentiation.
    This study evaluated the effects of nandrolone associated with resistance training (RT) on cardiac cytokines, angiotensin-converting enzyme activity (ACEA), and the sensitivity of the Bezold-Jarisch reflex (BJR). Male Wistar rats were... more
    This study evaluated the effects of nandrolone associated with resistance training (RT) on cardiac cytokines, angiotensin-converting enzyme activity (ACEA), and the sensitivity of the Bezold-Jarisch reflex (BJR). Male Wistar rats were divided into 3 groups: CONT (received vehicle, no training); EXERC (RT: after one week of water adaptation, rats were exercised by jumping into water twice a week for 4 weeks), and ND+EXERC (received nandrolone decanoate 10 mg/kg, twice/week, i.m, associated with RT). The BJR was analysed by measuring bradycardic and hypotensive responses elicited by serotonin administration. Myocyte hypertrophy and matrix collagen deposition were determined by morphometric analysis of H&E and picrosirius red-stained samples, respectively. TNF-α and ACEA were also studied. RT promoted physiological myocyte hyrpertrophy but did not cause changes in the other parameters. The association of ND with RT increased myocyte hypertrophy, deposition of matrix type I collagen, TNF-α and ACEA; decreased IL-10, and impairment in the BJR were observed in ND+EXERC compared with CONT and EXERC. ND is associated with alterations in cardiac structure and function as a result of the development of pathological cardiac hypertrophy (cardiac cytokine imbalance, elevation of ACEA) and cardiac injury, even when combined with resistance training.
    The objective of the present study was to employ high throughput image analysis to detect necrosis and apoptosis. Specific markers were replaced by morphological parameters of cells and nuclei. Fresh blood was taken from a healthy female... more
    The objective of the present study was to employ high throughput image analysis to detect necrosis and apoptosis. Specific markers were replaced by morphological parameters of cells and nuclei. Fresh blood was taken from a healthy female and given a treatment to induce cell necrosis and apoptosis. Afterward, the samples were stained with AnnexinV-FITC, DRAQ5 and DAPI. Slides were made and analyzed using the cytometer iCys. Pictures were scanned. The analyzed sample consisted of 73 sets of images of DAPI, DRAQ5 and AnnexinV-FITC, respectively. For image analysis and subsequent statistical processing, the CellProfiler and CellProfilerAnalyst were used. Each sample was analyzed twice. The first analysis was conducted using the markers (DAPI, DRAQ5 and Annexin) for an unequivocal identification and subsequent count of necrotic, apoptotic and live cells (gold standard). Thereafter, a second analysis was performed for the nuclear morphology and texture (morphometric analysis). After the m...
    ABSTRACT The rosemary (Rosmarinus officinalis L.) essential oil is widely used as an ingredient in food formulations and fragrance industry. However, there are few studies on the effects of seasonal variation on the composition and... more
    ABSTRACT The rosemary (Rosmarinus officinalis L.) essential oil is widely used as an ingredient in food formulations and fragrance industry. However, there are few studies on the effects of seasonal variation on the composition and biological activities of this oil. Thus, the aim of present study was to evaluate the effects of seasonal variation (2012–2013) on composition and antioxidant and antimicrobial activities of rosemary essential oils and ethanolic extract. Composition of essential oil was determined by GC-MS and that of the extract by LC-MS/MS. Antioxidant activity was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and beta-carotene/linoleic acid methods. Total phenolic compounds were determined by Folin–Ciocalteu method, and antimicrobial activity was evaluated by the determination of the minimum inhibitory concentration. Camphor was found as the major compound in essential oil (24.3–35.9%, mainly in January/2013), while carnosic acid was found in largest amount in January/2013 (summer) in the ethanolic extract, which presented the largest antioxidant activity, similar to butylated hydroxytoluene (BHT) and alfa-tocopherol. The results showed that essential oil and extracts harvested in summer exhibited strong activity against Staphylococcus aureus, even higher than sulfanilamide, due to the high amount of carnosic acid and camphor. Altogether, the essential oil and the extracts collected in summer showed better antioxidant and antimicrobial activities, which can be explained by the high levels of camphor and carnosic acid.
    An efficient and economical synthesis of the naturally occurring aromatase inhibitor abyssinone II was performed. The synthesis features an optimized aromatic prenylation reaction in which an arylcopper intermediate is reacted with prenyl... more
    An efficient and economical synthesis of the naturally occurring aromatase inhibitor abyssinone II was performed. The synthesis features an optimized aromatic prenylation reaction in which an arylcopper intermediate is reacted with prenyl bromide to afford a key intermediate that was converted to a prenylated aromatic aldehyde. Condensation of the aldehyde with an o-hydroxyacetophenone under Claisen-Schmidt conditions afforded a chalcone that was deprotected and cyclized in the presence of sodium acetate in refluxing ethanol to afford (+/-)-abyssinone II. The synthesis proved to be versatile enough to provide an array of abyssinone II derivatives that were evaluated as aromatase inhibitors. Methylation of the 4'-hydroxyl group of (+/-)-abyssinone II resulted in a significant increase in aromatase inhibitory activity, and further smaller increases in activity resulted from the methylation of the 7-hydroxyl group and removal of the prenyl side chain. As a result of these structural changes, the most active flavanone of the series was 20 times more potent than (+/-)-abyssinone II (IC50 40.95 microM).
    ABSTRACT The olive ridley turtle (Lepidochelys olivacea) is globally distributed. However, the species is considered vulnerable. Additionally, hematological and biochemical data for this species are seldom published in the scientific... more
    ABSTRACT The olive ridley turtle (Lepidochelys olivacea) is globally distributed. However, the species is considered vulnerable. Additionally, hematological and biochemical data for this species are seldom published in the scientific literature. The aim of this study was to gather hematological and biochemical data on young L. olivacea, drawing comparisons with hematological and biochemical data already published for sea turtles. Blood samples from 23 olive ridley sea turtles were collected, and hematological and biochemical parameters were determined. Mean and standard deviation values were determined for the hematological and biochemical results. From these results, a comparison with the published results for other turtles was performed using a two-way ANOVA and a Bonferroni post-test. The results of this study are consistent with previous findings in other sea turtles. No differences were found between genders. The results of the present study could be used as reference values by veterinarians in the treatment of olive ridley turtles as well as other species of sea turtles. There is a need to standardize the samples and analyses used to determine the parameters evaluated here to allow the determination of reference values for marine turtles and more comparable results.
    The aims of this study were to evaluate the effects of nandrolone (ND) on cardiac inflammatory cytokines, ACE activity, troponin I, and the sensitivity of the Bezold-Jarisch reflex (BJR). Male Wistar rats were administered either ND (20... more
    The aims of this study were to evaluate the effects of nandrolone (ND) on cardiac inflammatory cytokines, ACE activity, troponin I, and the sensitivity of the Bezold-Jarisch reflex (BJR). Male Wistar rats were administered either ND (20 mg/kg; DECA) or vehicle (control animals; CONT) for 4 weeks. BJR was analyzed by measuring the bradycardia and hypotension responses elicited by serotonin administration (2-32 μg/kg). Mean arterial pressure (MAP) was assessed and myocyte hypertrophy was determined by the heart weight/body weight ratio and by morphometric analysis. Matrix collagen deposition was assessed by histological analysis of the picrosirius red-stained samples. Mesenteric vascular reactivity was performed and central venous pressure (CVP) evaluated. Cardiac inflammatory cytokine levels and angiotensin-converting enzyme (ACE) activity were studied as well the biomarker of cardiac lesion, troponin I. DECA group showed enhancement of matrix type I collagen deposition (p < 0.01) and cardiac ACE activity (p < 0.01) compared with the CONT. Interleukin (IL)-10 was reduced (p < 0.01) and pro-inflammatory cytokines (TNF-α and IL-6; p < 0.01) were increased in the DECA group compared with CONT. Cardiac injury was observed in the DECA group shown by the reduction in cardiac troponin I (p < 0.01) compared with the CONT group. Animals in the DECA group also developed myocyte hypertrophy and reduction of BJR sensitivity. The MAP of animals treated with ND reached hypertensive levels (p < 0.01; compared with CONT). No changes in CVP and vascular reactivity were observed in both experimental groups. We conclude that high doses of ND elicit cardiotoxic effects with cardiac remodelling and injury. Cardiac changes reduce the BJR sensitivity. Together, these abnormalities contributed to the development of hypertension in animals in the DECA group.
    The objectives were to evaluate clinical and histopathological aspects of topical application of sunflower-seed oil (Helianthus annuus) on the healing process of experimentally induced wounds in lumbar and metacarpal areas of healthy... more
    The objectives were to evaluate clinical and histopathological aspects of topical application of sunflower-seed oil (Helianthus annuus) on the healing process of experimentally induced wounds in lumbar and metacarpal areas of healthy horses. Six adult horses were used. Eight wounds were surgically produced on each horse: two 6.25-cm2 wounds cranial to the sacrum on each side of the lumbar region
    The aim of this study was to expose the Yellow-spotted River Turtle, Podocnemis unifilis, to dietary cadmium (Cd) contamination. The P. unifilis were fed with a Cd contaminated diet (590 µgg(-1)) or a control diet for 30 and 60 days.... more
    The aim of this study was to expose the Yellow-spotted River Turtle, Podocnemis unifilis, to dietary cadmium (Cd) contamination. The P. unifilis were fed with a Cd contaminated diet (590 µgg(-1)) or a control diet for 30 and 60 days. After the Cd feeding period, the locomotor performance and specific growth rate were assessed. Blood samples were drawn for micronuclei analysis and tissues were collected to analyze the Cd concentration. Dietary Cd influenced the fitness of turtles at 30 days (righting time 752s), but not after 60 days (righting time 43.67s). Micronuclei in erythrocytes (12 ± 5‰) were significantly greater in contaminated turtle at 60 days. Cd accumulation is found in gut, intestine, kidney, fat, liver and blood of animals from contaminated diet group and the Cd concentration of almost all the tissues had increased following the 30-60-day feeding period. Cd does not impair animal the fitness after sixty days of dietary treatment, but it does can cause an accumulation on P. unifilis.
    ABSTRACT The production of counterfeit drugs is a criminal problem that carries serious risks to public health worldwide. Herein, the chemical fingerprinting of blister packaging using Fourier transform infrared spectroscopy (FTIR) of... more
    ABSTRACT The production of counterfeit drugs is a criminal problem that carries serious risks to public health worldwide. Herein, the chemical fingerprinting of blister packaging using Fourier transform infrared spectroscopy (FTIR) of authentic and counterfeit samples of Viagra® and Cialis® is demonstrated. Fifteen commercial samples (Viagra® and Cialis®) and thirty two counterfeit samples (Viagra and Cialis) were analyzed, and the FTIR data was subjected to chemometric treatment via unsupervised pattern recognition methods (principal component analysis, and hierarchical cluster analysis) and a supervised pattern recognition method (partial least squares discriminant analysis). ATR-FTIR spectra of the blister packaging of authentic Cialis® and counterfeit Cialis samples showed bands at 2976, 2904, 1431, 1326, 1243, 973, 691 and 608 cm−1, suggesting the presence of polyvinyl chloride (PVC) in its chemical composition. For authentic Viagra® and counterfeit Viagra samples, several distinct chemical profiles were observed in the ATR-FTIR spectra. Using unsupervised methods, samples were separated into three large groups: (i) counterfeit Viagra (seven samples made of PVC); (ii) authentic Viagra® (three samples made of poly(ethylene terephthalate)); (iii) Cialis (authentic and counterfeit) and some samples of Viagra (thirty seven made of PVC with additives of stearic acid derivatives, butyl hydroxy toluene or bisphenol A). Therefore, this suggests that three different types of forming films are used in the market for blister packaging used to contain inhibitors of PDE-5. Using supervised methods, all samples were correctly classified into their respective classes.
    ABSTRACT Fifteen carbohydrates (d-mannose, d-glucose, d-galactose, methyl-α-d-glucose, l-rhamnose, d-xylose, d-fructose, d-arabinose, dulcitol, mannitol, β-maltose, α-lactose, melibiose, sucrose, and raffinose) and four cyclitols... more
    ABSTRACT Fifteen carbohydrates (d-mannose, d-glucose, d-galactose, methyl-α-d-glucose, l-rhamnose, d-xylose, d-fructose, d-arabinose, dulcitol, mannitol, β-maltose, α-lactose, melibiose, sucrose, and raffinose) and four cyclitols [l-(+)-bornesitol, myo-inositol, per-O-acetyl-1-l-(+)-bornesitol, and quinic acid] were assayed for in vitro ACE inhibition. Of these molecules, per-O-Acetyl-1-l-(+)-bornesitol, quinic acid, methyl-α-d-glucose, d-rhamnose, raffinose, and the disaccharides were determined to be either inactive or weak ACE inhibitors, whereas l-(+)-bornesitol, d-galactose, d-glucose, and myo-inositol exhibited significant ACE inhibition. Molecular docking studies were performed to investigate interactions between active compounds and human ACE (Protein Data Bank, PDB 1O83). The results of various calculations showed that all active sugars bind to the same enzyme region, which is a tunnel directed towards the active site. With the exception of myo-inositol (K i = 13.95 μM, IC50 = 449.2 μM), the active compounds presented similar K i and IC50 values. d-Galactose (K i = 19.6 μM, IC50 = 35.7 μM) and l-(+)-bornesitol (K i = 25.3 μM, IC50 = 41.4 μM) were the most active compounds, followed by d-glucose (K i = 32.9 μM, IC50 = 85.7 μM). Our docking calculations are in agreement with the experimental data and show a new binding region for sugar-like molecules, which may be explored for the development of new ACE inhibitors.

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