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Made available in DSpace on 2021-06-25T11:13:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 This study evaluated the influence of Doxycycline (DOX) and Low-Intensity Laser (LIL) (InGaAlP diode laser) as scaling and root planing (SRP) adjuvants in the treatment of periodontitis induced in rats. The rats received periodontal disease induction, and after 7 days, the ligature was removed, and the animals were divided into groups/treatments: NT—receive no treatment; SRP—submitted only to SRP; DOX—submitted to SRP and DOX irrigation; LIL—submitted to SRP and LIL irradiation; and DOX + LIL—submitted to SRP treatments, DOX irrigation and LIL irradiation. The animals were sacrificed at 7, 15 and 30 days, and then, the analyses were performed. A lower concentration of Alpha-glycoprotein acid and Complement 3 was observed in the DOX + LIL group compared to all the other groups in all the periods, and for Complement 4 at 15 and 30 days (P

This study aims to evaluate the effects of photobiomodulation (PBM) on human monocytes, assessing the oxidative burst and ultimate fungicidal potential of these cells, as well as the gene expression at the mRNA level of CD68, CD80, CD163, CD204, IL-6, TNF-α and IL-10 in derived macrophages. Primary cultures of human monocytes were irradiated with an InGaAlP (660 nm)/GaAlAs (780 nm) diode laser (parameters: 40 mW, 0.04 cm2, 1 W/cm2; doses: 200, 400 and 600 J/cm2). Cells were submitted to the chemiluminescence assay, and a microbicidal activity assay against Candida albicans was performed. Reactive oxygen species (ROS) and nitric oxide (NO) production were measured, and cell viability was assessed by the exclusion method using 0.2% Trypan blue reagent. Irradiated monocytes were cultured for 72 h towards differentiation into macrophages. Total RNA was extracted, submitted to reverse transcription and real-time PCR. The results were analysed by ANOVA and the Tukey test (α = 0.05). Irradiated monocytes revealed a significant increase in their intracellular and extracellular ROS (P

Different Passiflora species have been appointed as a promising herbal medicine due to antioxidant properties; however, their effect on oxidative process induced by diabetes is still controversial. We aimed to evaluate effects of hydroethanolic extract 70% from P. edulis leaf on biochemical blood markers, collagen glycation, production of oxidant species and platelet aggregation in diabetic rats. The phytochemical analysis of the extract was performed by dereplication using LC coupled to the Photodiode Array Detector and Mass Spectrometer detector. Male Wistar rats were assigned to the control group and groups treated with alloxan (150 mg/kg) intraperitoneally, extract (200 mg/kg/d, for 90 d) and combination of alloxan and extract. The phytochemical analysis suggested the presence of flavonoids C-glycosides in the extract. The diabetic animals treated with the extract presented improvement in glycaemic control, reduced glycation collagen, levels of non-high density lipoprotein (non-HDL) cholesterol, total cholesterol and creatinine, production of oxidant species and aggregation in platelet in relation to diabetic animals non-treated. Our results showed that P. edulis leaf extract presents a health benefit to the diabetic state, preventing the appearance of its complications. Its effect can be associated with flavonoids, among which is the flavonoid C-glycoside isoorientin.

RESUMO Objetivo: Nesta revisão sistemática, nós avaliamos o link entre indutores de estresse oxidativo e/ou nitrosativo (EO/EN) com atividade antifúngica, através de uma ação direta sobre a célula fúngica e/ou modulando a resposta de fagócitos contra fungos de interesse médico (incluindo Candida spp., Cryptococcus spp. e Aspergillus spp.). Ainda, foram avaliadas as implicações clínicas deste evento bioquímico, bem como as perspectivas quanto à busca por novos compostos com atividade antifúngica, principalmente, os provenientes de fonte natural e, que explorem a indução de um EO ou EN como parte de seu mecanismo de ação. Metodologia: Foram avaliados artigos, provenientes de diferentes bases de dados e publicados a qualquer período, acessados entre abril e junho de 2017, através da utilização de diferentes descritores. Resultados: Primeiramente, estabelecemos as definições de EO/EN, como sendo o aumento das concentrações de espécies reativas do oxigênio e/ou nitrogênio (ERO/ ERN) quantificado diretamente e, provenientes de fontes fúngicas mitocondriais, Reação de Fenton, retículo endoplasmático ou outras não definidas, e excedendo a capacidade de defesa antioxidante do microrganismo (avaliados por análises de perfis transcriptômicos ou proteômicos ou metabolômicos ou níveis de atividade enzimática). Este aumento de ERO/ERN causando EO/EN é definido por tempo e condições que conduzem a sinalização de apoptose ou reais danos a biomoléculas com perda de função (peroxidação lipídica ou oxidação proteica ou danos ao DNA) e, consequentemente, gerando morte fúngica ou outro efeito antifúngico associado. Portanto, 64 artigos (apenas um publicado antes do ano 2000 e 50 entre 2007-2017) abordam que a indução de EO ou EN na célula fúngica é parte do mecanismo de ação de clássicos agentes antifúngicos (22 publicações), tais como azóis (fluconazol, itraconazol e miconazol), polienos (anfotericina B [AnB]) e equinocandinas (mica-fungina), assim como tal modulação redox tem sido reportada como um importante alvo terapêutico na busca por novos e promissores compostos naturais com atividade antifúngica (32 publicações), que tem respaldo pela grande variedade de indutores que podem provir da natureza. Ainda, compostos que também induzem o burst oxidativo de fagócitos, incluindo AnB, são potencializadores do efeito antifúngico in vivo. Além do efeito antifúngico contra células planctônicas, os efeitos dos EO ou EN sobre biofilmes fúngicos, também têm sido reportados. Tem sido firmado na literatura recente um claro link entre EO ou EN e a atividade antifúngica, tanto para aqueles agentes antifúngicos já utilizados na terapêutica em humanos, quanto para possíveis candidatos a fármaco. Portanto, a indução do EO ou EN como parte do mecanismo de ação de antifúngicos demonstra ser um importante alvo terapêutico, com perspectivas favoráveis sobre os desfechos na prática clínica. resumen está disponible en el texto completo SUMMARY Aim: In this systematic review, we evaluated the link between inducers of oxidative or nitrosative stresses (OS/NS) and antifungal activity against fungi of medical relevance (including Candida spp., Cryptococcus spp., and Aspergillus spp.), through a direct action on the fungal cell or modulating phagocyte response. Moreover, the clinical implications of this biochemical event, as well as the perspectives, were examined, highlighting the search for new compounds with antifungal activity, mainly those from natural sources and, which explores the induction of OS or NS as part of the mechanism of action. Methodology: Articles from different databases and published at any time were evaluated, between April and June 2017, and using different descriptors. Results: First, a definition of OS and NS was established in which an increase in reactive oxygen or nitrogen species (ROS/ RNS, quantified directly and from mitochondrial, Fenton reaction, endoplasmic reticulum or other fungal sources) should exceed the antioxidant defense capacity of the microorganism (evaluated by transcriptomic or proteomic or metabolomic profiles or enzyme activity levels). These events, by time and conditions delimited, can lead to the signaling of apoptosis or an actual damage toward biomolecules (lipid peroxidation or protein oxidation or DNA damage) and, consequently, they can cause cell death or other associated antifungal effect. Therefore, 64 articles were found, of these, only one was published before 2000 and 50 between 20072017, reporting the induction of OS or NS directly into the fungal cell via an increase in ROS or RNS as part of the mechanism of action of classical antifungal agents (22 publications), such as: azoles (fluconazole, itraconazole, and miconazole), polyenes (amphotericin B, [AnB]), and echinocandins (micafungin). This redox modulation has also been reported as an important therapeutic target in the search for new natural compounds with antifungal activity (32 publications), which is supported for the great variety of inducers from nature. Compounds that also induce the oxidative burst of phagocytes, including AnB, promote a combinatorial antifungal effect in vivo. In addition to the antifungal effect against plank-tonic cells, the relation between OS or NS and antifungal activity against fungal biofilms has also been reported. It has been established in the recent literature a clear link between OS or NS and antifungal effect, during the action of anti-fungal agents already used in the therapy in humans as well as for possible drug candidates. Thus, the induction of OS or NS as part of the mechanism of action proves to be an important therapeutic target with favorable perspectives on the outcomes in clinical practice.

Objective: To evaluate the pulp tissue of rat molars after pulpotomies with mineral trioxide aggregate (MTA), Biodentine TM (BDT) and calcium hydroxide (CH) mixed with sterile saline solution (24 hours, 72 hours, 7 days and 15 days), through correlating MPO activity with active neutrophils and MMP8 activity with tissue remodeling . Material and Methods: Thirty-eight Wistar rats were randomly distributed into groups (control, I (MTA gray), II (BDT), and III (CH)) and subdivided according to the study period of 24 hours, 72 hours, 7 days or 15 days after pulpotomy. MMP8 activity was assessed through fluorescence technique, and MPO activity was determined using the MPO assay. Results: A gradual decrease of MPO and MM8 activity occurred in the group MTA over the experimental periods (p

Made available in DSpace on 2022-05-01T09:47:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Purpose: This study evaluated the systemic and local effects of doxycycline (DOX) and lowintensity laser (LIL) treatment as adjuvants to scaling and root planing (SRP) in the treatment of experimental periodontitis in rats. Methods: The sample consisted of 180 male rats (Rattus norvegicus albinus, Wistar), of which 30 did not receive induction of periodontal disease (negative control [NC] group) and 150 received induction of periodontal disease in the lower first molar. After 7 days, the ligature was removed, and the animals were divided into the following groups: NT (no treatment), SRP (SRP), DOX (SRP and DOX irrigation), LIL (SRP and laser irradiation), and DOX+LIL (SRP, DOX, and LIL). The animals were euthanized at 7, 15, and 30 days; thereafter, biochemical, radiographic, histological, and immunohistochemical analyses were performed. Results: In the intragroup analysis, lower concentrations of α-1-glycoprotein acid (α-1-Ga) and complement 3 (C3) were observed in the DOX+LIL group than in all other groups at all time points, as well as lower levels of complement 4 (C4) at 15 and 30 days (P

This study aims to evaluate the effects of photobiomodulation (PBM) on human monocytes, assessing the oxidative burst and ultimate fungicidal potential of these cells, as well as the gene expression at the mRNA level of CD68, CD80, CD163, CD204, IL-6, TNF-α and IL-10 in derived macrophages. Primary cultures of human monocytes were irradiated with an InGaAlP (660 nm)/GaAlAs (780 nm) diode laser (parameters: 40 mW, 0.04 cm

espanolObjetivos: este estudio investigo la bioactividad del extracto de hoja en bruto (EHB) y las fracciones hexano (HX) y acetato de etilo (AcOEt) de Talinum paniculatum solo y en asociacion con fluconazol (FLC) contra cepas de referencia y aislados clinicos de Candida albicans resistente a FLC. Ademas, evaluo la capacidad antioxidante, la composicion quimica de esta planta y los mecanismos subyacentes del efecto fungicida. Metodos: la actividad antifungica se evaluo mediante microdilucion en caldo para establecer las concentraciones inhibitorias minimas (CIM) y microbicidas minimas (CMM). Durante el tratamiento con FLC y productos vegetales se detecto la generacion de especies reactivas de oxigeno (ERO) (radicales hidroxilo [HO•]) en celulas de C. albicans utilizando las sondas fluorescentes permeables a la membrana APF y HPF. El perfil de cromatografia liquida de alta resolucion (CLAR), el analisis cuantitativo de compuestos antioxidantes y el ensayo DPPH fueron evaluados. Resultados: el EHB y las fracciones presentaron una excelente actividad antifungica y selectividad contra las celulas de C. albicans, pero no tuvieron efectos sinergicos con FLC. Los valores de CIM para EHB y sus fracciones contra la cepa referencia de C. albicans fueron del orden de: HX (31,25 μg ml–1) EnglishAims: This study investigated the bioactivity of the crude leaf extract (CLE) and fractions hexane (HX) and ethyl acetate (EtOAc) from Talinum paniculatum alone and in association with fluconazole (FLC) against reference strain and clinical isolates of FLC-resistant Candida albicans. Furthermore, the antioxidant capability, chemical composition of this plant, and the effect’s underlying mechanisms were evaluated. Methods: The antifungal activity was evaluated using checkerboard assay to establish the minimum inhibitory (MIC) and minimum microbicidal concentrations (MMC). During FLC and plant products challenges, the reactive oxygen species (ROS) generation (hydroxyl radicals [HO•]) were detected in C. albicans cells using the membrane-permeable fluorescent probes APF and HPF. High-performance liquid chromatography (HPLC) profile, quantitative analysis of antioxidant compounds, and free radical scavenging activity (DPPH assay) tests were performed. Results: The CLE and fractions presented outstanding antifungal activity and selectivity against C. albicans cells but had no synergistic effects with FLC. The MIC values for CLE and its fractions against C. albicans reference strain were in the order of HX (31.25 μg ml–1)

Propose: We evaluated the antibacterial potential of the crude leaf extract (CLE) and fractions hexane (HX) and ethyl acetate (EtOAc) from Talinum paniculatum alone and in association with oxacillin (OXA) against OXA-resistant Staphylococcus aureus (ORSA, environment isolates) and OXA-sensitive S. aureus (OSSA, ATCC 25923). Furthermore, toxicity tests were performed. Methods: The antibacterial activity was evaluated through checkerboard assay (broth microdilution) to establish the minimum inhibitory (MIC) and minimum bactericidal concentrations (MBC). Toxicity test in mice was assessed. Results: The MIC values for the CLE and its fractions against ORSA and OSSA were in the order of HX (500 μg ml–1) = EtOAc < CLE (4000 μg ml–1). EtOAc and HX presented outstanding antibacterial activities against ORSA, and these fractions were bactericidal toward OSSA. Conversely, the associations between plant product (CLE, EtOAc, or HX) and OXA exhibited no synergistic effects. During these associations, there was an increase in OXA MICs anywhere from 2- to 4092-fold. The CLE presented absence of toxicity at a dose of 5 g kg-1 (in vivo). Conclusion: Although T. paniculatum be a good source of bioactive compounds with antistaphylococcal potential, the researchers should be cautious, since its edible leaf may interfere with OXA therapy (mitigating OXA-induced growth inhibition or killing of S. aureus and enhancing S. aureus resistance).

Background and aims: Lipopolysaccharide (LPS), an endotoxin, is a component of the outer membrane of Gram-negative bacteria that is able to activate the peripheral immune system, leading to changes in signalling pathways that act locally and systemically to achieve adaptive responses. Sickness behaviour is a motivational state in response to endotoxin exposure and includes depressed activity and a reduction of exploratory behaviour, potentially reorganising organism priorities to cope with infectious diseases. We hypothesised that 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (Tempol) modulates the leukocyte response to endotoxins and decreases LPS-induced sickness behaviour in mice.Methods: The effects of Tempol on LPS-induced peritonitis and the respiratory burst of neutrophils primed with LPS and triggered by phorbol 12-myristate-13-acetate (PMA) were evaluated. To evaluate the effects of Tempol on sickness behaviour, the mice were submitted to an open field and forced swim tests.Results: Tempol (50-100 μM/106 cells) decreased the respiratory burst of LPS-primed and PMA-stimulated neutrophils in vitro. In vivo, this nitroxide (30 and 100 mg/kg body weight) inhibited leukocyte migration to the peritoneal cavity after LPS administration in mice. Moreover, Tempol pretreatment (30 and 100 mg/kg body weight) before LPS administration also attenuated sickness behavioural changes.Conclusions: Together, these findings shed light on the mechanisms underlying the anti-inflammatory potential and confirm the therapeutic potential of nitroxides.

Aims The identification of novel targets to control inflammation in humans is probably the primary challenge that impairs the development of new anti-inflammatory drugs. Therefore, the modulation of intracellular signaling pathways in phagocytes may be an interesting means of achieving this goal. However, this change to signaling can compromise the host's susceptibility to invading pathogens. We investigated whether the antioxidant nitroxide Tempol regulates the activity of kinases associated with the production of oxidants in neutrophils, which affects the fungicidal capability of these cells. Main methods The effects of Tempol on PMA- or fMLP-activated neutrophils were examined by oxygen consumption as an index of the oxidative burst, a release of extracellular and total Reactive Oxygen Species (ROS) by chemiluminescence, kinase activities through analysis of ATP consumption during enzyme activities and the dot blot immunoassay and, finally, by neutrophil capacity of killing Candida albicans. Key findings Tempol significantly inhibited the neutrophil oxidative burst in a concentration-dependent manner and decreased oxygen consumption (IC50 = 45 μM) and extracellular/total ROS formation with an increase on the lag period response. In addition, Tempol inhibited neutrophil kinase activities (i.e., a decrease in protein phosphorylation) elicited through different biochemical pathways and consequently impaired the fungicidal activity of these cells. Significance Although Tempol has potential anti-inflammatory activity that acts on different intracellular pathways (such as those involving kinases), researchers should be cautious, since this nitroxide down-regulated oxidants production and the fungicidal response of neutrophils.

Potential endpoint biomarkers were evaluated in the assessment of exposure to triazoles, in the southern region of Minas Gerais, Brazil. Volunteers were divided into three groups: occupationally exposed and rural residents (n = 21), non-occupationally exposed and rural residents (n = 35) and non-occupationally exposed and urban residents (n = 30). Of all endpoints evaluated, plasma concentration of androstenedione (p < 0.001) and glycine-conjugated bile acids presented statistical differences in the three studied groups (p < 0.05). However, our findings concerning oxidative stress and testosterone levels, plus that related to unconjugated and taurine conjugated bile acids, suggested that more studies are necessary to evaluate their potential as biomarkers for triazole exposure, as statistical significance was not attained between the groups. Our human population data contributes to the development of triazole exposure risk assessment with respect to these potential effect biomarkers, in potentially vulnerable groups and individuals.

Introduction and Objective: Dyslipidemia is associated with increased risk of cardiovascular disease and atherosclerosis, and hence with high morbidity and mortality. This study investigated the effects of the nitroxide 4-hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl (Tempol) on lipid profile and cardiac morphology in low-density lipoprotein (LDL) receptor gene knockout (LDLr-/-) mice. Methods: Male LDLr-/- mice (three months old, approximately 22 g weight) were divided into the following groups: controls, including (1) standard chow (SC, n=8) and (2) high-fat diet (HFD, n=8); and treatment, including (3) standard chow + Tempol (SC+T, n=8) (30 mg/kg administered by gavage, once daily) and (4) high-fat diet + Tempol (HFD+T, n=8) (30 mg/kg). After 30 days of the diet/treatment, whole blood was collected for analysis of biochemical parameters (total cholesterol, triglycerides [TG], high-density lipoprotein [HDL], LDL, and very low-density lipoprotein [VLDL]). The heart was removed through thoracotomy and histological analysis of the left ventricle was performed. Results: A significant increase in TG, LDL, and VLDL and marked left ventricular hypertrophy (LVH) were demonstrated in the HFD group relative to the SC group (p

Low intensity laser can be used as a promising alternative in the treatment of periodontal disease. Objective The aim of this study was to evaluate low-level laser therapy (LLLT) as an adjuvant treatment for scaling and root planing (SRP) for the treatment of induced periodontitis in simvastatin-modified rats. Material and Methods A total of 180 rats were evenly divided into two groups: Veh – receiving oral administration of polyethylene glycol (vehicle); S – receiving oral administration of Simvastatin. Periodontal disease was induced in both groups at the first mandibular molar. After seven days, the ligature was removed and the animals were divided into subgroups according to the following local treatments: NT – no treatment; SRP – scaling and root planing and irrigation with saline solution; and LLLT ¬– SRP and laser irradiation (660 nm; 0.03 W; 4 J). Ten animals in each subgroup/local treatment were euthanized at 7, 15 and 30 days. Samples of gingival tissue were processed to analyze the tissue oxidative damage and radiographic analysis. Levels of oxidative stress were analyzed by the expressions of Tripeptideglutathione (TG), Malondialdehyde (MDA) and Carbonylated Proteins (CP). Results The animals in S group had higher levels of TG and lower levels of MDA and CP compared with Veh group (p

Background and aim Oxidative stress arising from inflammatory processes is a serious cause of cell and tissue damage. Tempol is an efficient antioxidant with superoxide dismutase-like activity. The purpose of this paper is to address the inhibition of protein disulfide isomerase (PDI), an essential redox chaperone whose active sites contain the Cys-Gly-His-Cys (CXXC) motif, by the nitroxide Tempol. Results In the presence of Tempol (5–120 μM), the reductase activity of PDI was reversibly affected both in vitro and in activated mice neutrophils, with an IC50 of 22.9 ± 10.8 μM. Inhibitory activity was confirmed by using both the insulin method and fluorescent formation of eosin-glutathione (E-GSH). The capacity of Tempol to bind the enzyme was determined by EPR and mass spectrometry. EPR Tempol signal decreased in the presence of PDI while remained unaffected when PDI thiols were previously blocked with NEM. When total protein was analyzed, 1 and 4 molecules of Tempol were bound to the protein. However, only one was found to be covalently bound to PDI at the a′active site. More specifically, Cys400 was modified by Tempol. Conclusion We have shown that the nitroxide Tempol acts as an inhibitor of PDI through covalent binding to the Cys400 of the protein structure. Since PDI is coupled with the assembly of the NADPH oxidase complex of phagocytes, these findings reveal a novel action of Tempol that presents potential clinical applications for therapeutic intervention to target PDI knockdown in pathological processes in which this protein is engaged.

Objective The objective of this study was to evaluate the local effects of statins as adjuvants for treatment by scaling and root planing (SRP) of periodontal disease induced in rats. Material and Methods Ninety rats were used in the present experiment. Periodontal disease was induced in all animals using a cotton thread placed in the left first mandibular molar. After 7 days of induction, the bandage was removed and the animals were divided into three groups: 1) NT group (n=30), no treatment; 2) SRP group (n=30): SRP and irrigation with control gel; 3) S group (n=30) - SRP and irrigation with Simvastatin. Ten animals from each group were euthanized at 7, 15 and 30 days after treatment. Gingival biopsy specimens were processed to analyze the expression of matrix metalloproteinase 8 (MMP-8). The mandibles were removed and submitted to radiographic and laboratory processing for histometric analysis. Results The S group showed a significantly lower expression of MMP-8 compared to NT and SRP groups in all experimental periods. In the radiographic and histometric analyses between the groups, S group showed a significantly lower bone loss (BL) compared to NT and SRP groups in all experimental periods. Conclusions Within the limits of this study, it can be concluded that locally applied statin was effective as an adjuvant treatment for SRP in rats with induced periodontal disease.

Different Passiflora species have been appointed as a promising herbal medicine due to antioxidant properties; however, their effect on oxidative process induced by diabetes is still controversial. We aimed to evaluate effects of hydroethanolic extract 70% from P. edulis leaf on biochemical blood markers, collagen glycation, production of oxidant species and platelet aggregation in diabetic rats. The phytochemical analysis of the extract was performed by dereplication using LC coupled to the Photodiode Array Detector and Mass Spectrometer detector. Male Wistar rats were assigned to the control group and groups treated with alloxan (150 mg/kg) intraperitoneally, extract (200 mg/kg/d, for 90 d) and combination of alloxan and extract. The phytochemical analysis suggested the presence of flavonoids C-glycosides in the extract. The diabetic animals treated with the extract presented improvement in glycaemic control, reduced glycation collagen, levels of non-high density lipoprotein (non-HDL) cholesterol, total cholesterol and creatinine, production of oxidant species and aggregation in platelet in relation to diabetic animals non-treated. Our results showed that P. edulis leaf extract presents a health benefit to the diabetic state, preventing the appearance of its complications. Its effect can be associated with flavonoids, among which is the flavonoid C-glycoside isoorientin.

Oxidative stress is an important risk factor for cardiovascular disease and death in hemodialysis (HD) patients. However, whether biochemical and nutritional markers might be useful to stratify HD patients according to the risk of oxidative damage remains unclear. We investigated whether low-cost and easily available parameters such as the profile of nutrients intake, nutritional status, and antioxidant defenses can predict lipid and protein oxidation in HD patients. Forty-nine HD patients (women=20,men=29), ranging from 18 to 65 years of age (73.5%) were submitted to biochemical and nutritional analysis. At least 93.9% of HD patients had malnutrition. A patient’s stratification according to nutritional risk was highly coherent with anthropometric parameters and nutrients intake, which were complementarily used as markers of malnutrition. Nutritional stratification was unable to reveal differences in the oxidative status. On the other hand, carbohydrate and zinc intake, serum zinc (Zn), glutathione peroxidase (GPx) activity, total antioxidant capacity (TAC), and nonprotein antioxidants (npAC) in serum were predictive markers of lipid (R2=0.588,P<0.001) and protein (R2=0.581,P<0.001) oxidation. Interestingly, GPx activity, TAC, and npAC exhibited good (>80%<90%) or excellent (>90%) accuracy to estimate lipid oxidation (P≤0.01). Regarding the prediction of protein oxidation, GPx activity and TAC presented regular accuracy (>70%<80%), and Zn serum levels exhibited good sensitivity (P≤0.01). Herein, we provided evidence that clinical characteristics relevant to predict different levels of lipid and protein oxidation in HD patients can be easily obtained, during routine hospital visits by means of the combined analyses of biochemical and nutritional parameters.

Secondary disorders in consequences to diabetes involves the development of several diseases in the oral cavity, as periodontitis, xerostomy, infection by diverse pathogens and dysfunctions on the salivary secretion. These alterations occur partially, in consequence of the oxidative stress occasioned by hyperglycemia, and are important in patients undiagnosed or that have flaws in their therapeutic process. The aim of this work was to evaluate biochemical alterations of submandibular glands in response to oxidative stress during diabetes mellitus, and verify the effects of N-acetylcystein supplementation to diabetic rats, specially on the regulation of modifications related to glutathione and thiol proteins. For this purpose, the levels of some oxidative stress markers and the occurrence of the post-translational event of S-glutathionylation were evaluated. The a-amilase degranulation by isolated acinar cells and glandular relative weight was also measured for each experimental group. The compound was able to decrease the lipoperoxidation and proteic oxidation observed in the submandibular gland of diabetic rats, preventing the decrease of the tecidual reducing power and increasing the occurrence of the post-translational process of S-glutathionylation. The diabetic condition increases the degranulation of a-amilase and the glandular weight, but the supplementation with N-acetylcystein did not affect these events. Together these findings may help to elucidate the status of oxidative stress on salivary glands and suggest new therapeutic strategies employing antioxidants of low molecular weight to prevent oral and systemic dysfunctions related to diabetes. Los trastornos secundarios que resultan de la diabetes implican el desarrollo de diversas enfermedades en la cavidad bucal, como periodontitis, xerostomía, infección por diversos patógenos y disfunciones en la secreción salival. Estos cambios ocurren parcialmente, debido al estrés oxidativo causado por la hiperglucemia, y son importantes en pacientes que no están diagnosticados o tienen fallas en el proceso terapéutico. El objetivo de este trabajo fue evaluar las alteraciones bioquímicas de las glándulas submandibulares en respuesta al estrés oxidativo durante la diabetes mellitus, y verificar la combinación farmacológica de la suplementación con N-acetilcisteína en ratas diabéticas, especialmente en la regulación de cambios relacionados con el glutatión. y proteínas de tiol. Para ello, se evaluaron los niveles de algunos marcadores de estrés oxidativo y la ocurrencia del evento de S-glutatilación postraduccional. También se midieron la desgranulación de la a-amilasa por las células acinares aisladas y el peso relativo de la glándula para cada grupo experimental. El compuesto fue capaz de disminuir la lipoperoxidación y oxidación proteica observada en la glándula submandibular de ratas diabéticas, evitando la reducción del poder reductor tisular y aumentando la ocurrencia del proceso postraduccional de S-glutatilación. La condición diabética aumenta la desgranulación de a-amilasa y el peso glandular, pero la suplementación con N-acetilcisteína no afectó estos eventos. Juntos, estos hallazgos pueden ayudar a dilucidar el estado de estrés oxidativo en las glándulas salivales y sugerir nuevas estrategias terapéuticas que utilizan antioxidantes de bajo peso molecular para prevenir los trastornos orales y sistémicos relacionados con la diabetes. Desordens secundárias em consequências da diabetes envolvem o desenvolvimento de diversas doenças na cavidade oral, como periodontite, xerostomia, infecção por diversos patógenos e disfunções na secreção salivar. Essas alterações ocorrem parcialmente, em decorrência do estresse oxidativo ocasionado pela hiperglicemia, e são importantes em pacientes não diagnosticados ou que apresentam falhas no processo terapêutico. O objetivo deste trabalho foi avaliar as alterações bioquímicas das glândulas submandibulares em resposta ao estresse oxidativo durante o diabetes mellitus, e verificar o conjunto farmacológico da suplementação de N-acetilcisteína em ratos diabéticos, especialmente na regulação das modificações relacionadas às proteínas glutationa e tiol. Para tanto, foram avaliados os níveis de alguns marcadores de estresse oxidativo e a ocorrência do evento pós-translacional de S-glutationilação. A degranulação da a-amilase por células acinares isoladas e o peso relativo glandular também foram medidos para cada grupo experimental. O composto foi capaz de diminuir a lipoperoxidação e oxidação protéica observadas na glândula submandibular de ratos diabéticos, evitando a diminuição do poder redutor tecidual e aumentando a ocorrência do processo pós-translacional de S-glutationilação. A condição diabética aumenta a degranulação da a-amilase e o peso glandular, mas a suplementação com N-acetilcisteína não afetou esses eventos. Juntos, esses achados podem ajudar a elucidar o estado do estresse oxidativo nas glândulas salivares e sugerir novas estratégias terapêuticas empregando antioxidantes de baixo peso molecular para prevenir disfunções orais e sistêmicas relacionadas ao diabetes.

The purpose of this study was to assess, for the very first time, the effects of photodynamic therapy (PDT) on Schistosoma mansoni in vitro by measuring reactive oxygen species (ROS) generation throughout the treatment, as well as the behavior of the parasites (mating, motility and contraction/shortening), and damage to their tegument and excretory systems. The parasites were divided into 4 groups: control, photosensitizer, laser and PDT. Light irradiation was delivered with an InGaAlP low-level laser device operating at 660nm, with 40mW and 100J/cm2. For PDT, different toluidine blue dye (TBO) concentrations and times of exposure were utilized. Interestingly, TBO-mediated PDT was able to kill S. mansoni (P

While researchers have extensively evaluated the beneficial effects of coffee consumption in reducing the frequency of certain diseases, studies examining the differences between organic and conventional coffee intake are still needed. Therefore, this paper aims to investigate the functional effects of organic and conventional coffee by examining both its chemical composition and its mutagenic/antimutagenic properties. Infusions of 10% or 20% (w/v) of organic and conventional coffee were administered by gavage (10 mL/kg b.w., once or twice a day) to male Swiss mice against doxorubicin (DXR) and 1,2-dimethylhydrazine dihydrochloride (DMH)-induced mutagenicity. The levels of chlorogenic acids, caffeine and trigonelline from the coffee infusions and oxidative stress analysis from the liver were measured by HPLC. Gut and bone marrow micronucleus assays were used as mutagenic/antimutagenic endpoints, as well as the crypt measurements and gut apoptosis index. The in vivo tests revealed that only organic coffee exerted protective effects, despite oxidative stress analysis and crypt measurements not showing differences among treatments. Intriguingly, the low dose (10% w/v mL/kg) displayed a robust protective effect that showed a significant reduction in bone marrow micronuclei (26.8%), gut micronuclei (11.5%) and apoptosis (27.8%), whereas the higher coffee dose (2 × 20% w/v) only showed a protective effect against bone marrow micronucleus (43.7%). These results highlight that organic coffee could be considered to have beneficial functional effects, although it is still a challenge to define conclusions from analytical data and all the possible interactions from this complex food matrix.

OBJECTIVE: The study presents the results of a 90-day safety assessment of rats fed with four varieties of soybeans, BRS 245 RR and BRS Valiosa RR (transgenic), BRS 133 and MG BR46 Conquista (non-transgenic). METHODS: Diets were prepared by incorporating toasted soybean flour to a commercial diet at 1%, 10% or 20% weight In the in vivo experimental the rats' body weight, body weight gain, food consumption, number of aberrant crypt foci, oxidative stress biomarkers, urea and creatinine levels were analyzed and compared between experimental groups, as well as histopathological observations (digestive tract, liver, kidneys). RESULTS: The results indicate that glyphosate-tolerant soy varieties neither induce nor prevent aberrant crypt foci induction, nor do their conventional counterparts. Similarly, none of the four soybean varieties tested induced changes in the digestive tract, liver or kidney. Serum biochemical parameters were also unchanged. CONCLUSION: The consumption of both, conventional and transgenic soybeans, were insufficient to ameliorate dimethylhydrazine-induced oxidative stress. OBJETIVO: Este estudo apresenta os resultados de um experimento de 90 dias com o objetivo de avaliar a segurança de quatro variedades de grãos de soja: BRS 245 RR e BRS Valiosa RR (transgênicas), BRS 133 e MG BR46 Conquista (não transgênicas). MÉTODOS: As dietas foram preparadas incorporando farinha de grãos de soja à dieta comercial (FRI-LAB Ratos II) a 1%, 10% ou 20% m/m. O peso corpóreo dos animais, o ganho de peso, o consumo de dieta, o número de focos de criptas aberrantes e os níveis de marcadores de estresse oxidativo, de creatinina e de ureia foram comparados entre os grupos experimentais, assim como as observações histopatológicas (trato digestivo, fígado e rins). RESULTADOS: Os resultados indicaram que as variantes glifosato-tolerantes não induziram ou preveniram a indução de focos de criptas aberrantes, assim como suas parentais convencionais. Similarmente, nenhuma das quatro variedades de grãos de soja testadas induziu alterações no trato digestivo, no fígado e nos rins. Os parâmetros bioquímicos do soro permaneceram também inalterados. CONCLUSÃO: Tanto o consumo de grãos de soja convencionais quanto o de transgênicos foram ineficazes para melhorar os níveis de estresse oxidativo induzidos pela dimetilhidrazina.

Oxidative stress arising from inflammatory processes is a serious cause of cell and tissue damage. Tempol is an efficient antioxidant with superoxide dismutase-like activity. The purpose of this paper is to address the inhibition of protein disulfide isomerase (PDI), an essential redox chaperone whose active sites contain the Cys-Gly-His-Cys (CXXC) motif, by the nitroxide Tempol.In the presence of Tempol (5-120 μM), the reductase activity of PDI was reversibly affected both in vitro and in activated mice neutrophils, with an ICWe have shown that the nitroxide Tempol acts as an inhibitor of PDI through covalent binding to the Cys400 of the protein structure. Since PDI is coupled with the assembly of the NADPH oxidase complex of phagocytes, these findings reveal a novel action of Tempol that presents potential clinical applications for therapeutic intervention to target PDI knockdown in pathological processes in which this protein is engaged.

The antifungal drug therapy often employed to treat paracoccidiodomycosis (PCM), an important neglected fungal systemic infection, leads to offensive adverse effects, besides being very long-lasting. In addition, PCM compromises the oral health of patients by leading to oral lesions that are very painful and disabling. In that way, photodynamic therapy (PDT) arises as a new promising adjuvant treatment for inactivating Paracoccidioides brasiliensis (Pb), the responsible fungus for PCM, and also for helping the patients to deal with such debilitating oral lesions. PDT has been linked to an improved microbial killing, also presenting the advantage of not inducing immediate microbial resistance such as drugs. For the present study, we investigated the generation of reactive oxygen species (ROS) by using the fluorescent probes hydroxyphenyl fluorescein (HPF) and aminophenyl fluorescein (APF) after toluidine blue (TBO-37.5 mg/L)-mediated PDT (660 nm, 40 mW, and 0.04 cm2 spot area) and the action of TBO-PDT upon Pb cultures grown for 7 or 15 days in semisolid Fava Netto's culture medium; we also targeted oral PCM manifestations by reporting the first clinical cases (three patients) to receive topic PDT for such purpose. We were able to show a significant generation of hydroxyl radicals and hypochlorite after TBO-PDT with doses around 90 J/cm2; such ROS generation was particularly useful to attack and inactivate Pb colonies at 7 and 15 days. All three patients reported herein related an immediate relief when it came to pain, mouth opening, and also the ability to chew and swallow. As extracted from our clinical results, which are in fact based on in vitro outcomes, TBO-PDT is a very safe, inexpensive, and promising therapy for the oral manifestations of PCM.

O estudo avaliou a Sinvastatina no tratamento da doença periodontal (DP) em ratos. Trinta e seis animais foram divididos em Grupo Controle (C) que receberam oralmente soro fisiológico e Sinvastatina (S) que receberam oralmente Sinvastatina. Após 24 horas, a DP foi induzida utilizando-se um fio de algodão na região dento-gengival dos primeiros molares inferiores esquerdos. Após 7 dias, a ligadura foi removida e os animais receberam raspagem e alisamento radicular (RAR) e irrigação com soro fisiológico. Os animais foram eutanasiados aos 7, 15 e 30 dias. Radiograficamente, o tratamento com RAR mostrou uma menor perda óssea (PO) no Grupo S comparado ao C em todos os períodos. Concluiu-se que a Sinvastatina associada à RAR foi efetiva na redução da PO em ratos. Periodontal disease (PD) is a multifactorial disease whose main signs are the insertion loss of attachment and alveolar bone. It has been revealed that statins could stimulate bone formation. The objective of this study was to evaluate the effects of simvastatin in adjuvant treatment of PD. 36 rats were divided into two groups, Control, who received saline orally; and Simvastatin who received simvastatin. After 24 hours, the PD was induced in animals using a cotton thread in the first lower left molar. After 7 days, the animals were treated with scaling and root planing. They were euthanized at 7, 15, and 30 days, and radiographic analysis demonstrated less bone loss in animals treated with simvastatin.

Faced with the search for healthy products that provide additional benefits to consumers’ health, the main objectives of this work were to develop a low-calorie preserve containing prebiotics (lactulose and polydextrose) and to evaluate the effects of these prebiotics on oxidative stress and colon carcinogenesis in male rats treated with 1,2-dimethylhydrazine (DMH). A total of 62.5% w/w of the sucrose in conventional preserves was replaced by polydextrose, and lactulose was added at 0%, 16%, 19.5% or 23% w/w concentrations. The acceptance of these four low-calorie guava preserve samples and the conventional sample was equal ( P > 0.05), with a score of 6.49. The level of degradation of lactulose was low (18.45 g 100 g −1 lactulose), ensuring that even at a lower concentration of this prebiotic (16% w/w), the concentration remained above the minimum level considered functional. We found that consumption of the low-calorie preserves with prebiotics does not have an effect on the development of mucin-negative ACF and classical ACF in the initiation phase of the mutagenic process. However, the consumption of 1.5 g of the preserve/rat/day potentiated lipid peroxidation and proteic oxidation in the liver.

In the present study, we evaluated the pesticide and metal concentrations as well as the antimutagenic and mutagenic properties of commercial soybeans (Glycine max). Male Swiss mice were fed diets containing 1%, 10%, or 20% (w/w) transgenic soybeans (BRS Valiosa RR) or parental isogenic conventional soybeans (MG-BR46 Conquista). Cyclophosphamide (50 mg kg⁻¹ b.w.) was added in a single dose 24 h before euthanasia as an induction agent. There was no difference in the composition (ash, total fat, protein, moisture, and carbohydrates) of the diets containing the same soybean concentration. The results show that the commercially available Brazilian soybeans tested are free of organochlorine, organophosphate, and carbamate pesticides and contain acceptable heavy metal concentrations. Both cyclophosphamide and soybean treatments were not sufficient to cause detectable oxidative damage on liver by the levels of malondialdehyde and protein carbonyl. The transgenic soybeans are also nonmutagenic and have protective effects against DNA damage similar to those of conventional soybeans but to a lesser percentage (64%-101% for conventional and 23%-33% for transgenic diets).

Despite defenses by polymorphonuclear neutrophils in the host against invading agents, overproduction of oxidant species by phagocytes can lead to damage in the surrounding tissues. Several benzophenones have been shown to possess anti-inflammatory properties. The effect of the natural benzophenone 7-epiclusianone isolated from leaves of Garcinia brasiliensis was investigated by using in vitro antioxidant and ex vivo anti-inflammatory assays, focusing on the neutrophil respiratory burst and on the biochemical pathways involved. The bioactive extract, 7-epiclusianone, showed low in vitro antioxidant activity as evaluated by the 2,2-diphenyl-1-picrylhydrazyl free radical scavenging assay, the reducing power test, or the chelating power assay. However, the benzophenone displayed potent activity in the ex vivo model of the neutrophil respiratory burst, inhibiting the generation of superoxide anions in a dose-dependent manner. When the respiratory burst was triggered by N-formyl-methionyl-leucyl-phenylalanine, a chemotactic peptide, the 50% effective concentration (EC(50)) was 41.18 μg/10(7) cells. When phagocytes were stimulated directly through protein kinase C via phorbol, the EC(50) was 34.3 μg/10(6) cells. The results indicated that 7-epiclusianone was able to down-regulate inflammatory phagocyte superoxide anion release through a mechanism controlled by tyrosine protein phosphorylation and by a direct stimulation of protein kinase C. These findings could lead to new therapeutic approaches for inflammation management and the development of new drugs.

Differentiation between organic and conventional coffee has increased due to the growing demand and high consumption of healthy foods that contain compounds with antioxidant potential, which have been associated with the reduction of chronic diseases. We used organic and conventional coffee in powder 4% (w/w) and infusions 5%, 10% and 20% (w/v) incorporated in a commercial diet to test in vivo. The levels of chlorogenic acid, caffeine and trigonelline were determined by high performance liquid chromatography. The body weight, weight gain, food consumption, aberrant foci crypt, mucin depleted foci, stress biomarkers protein carbonyl and malondialdehyde, biochemical parameters and behavior of the rats were compared between the experimental and control groups within a framework of colon carcinogenesis. The organic coffee showed higher levels of chlorogenic acid, caffeine and trigonelline than conventional, however, this difference did not significantly affect behavior. The infusions had an antioxidant effect, reducing the levels of malondialdehyde; however, the biochemical parameters of the serum were not altered, and there was neither induction nor prevention of preneoplasic lesions.

The increased incidence of infections caused by the opportunistic pathogen Cryptococcus neoformans, which mainly affects immunocompromised patients but can also infect immunocompetent individuals, has needed additional studies on this micro-organism's pathogenicity and factors related to virulence, such as enzyme production, for a better understanding of the aetiology of cryptococcosis. The aim of this study was to verify the applicability of non-denaturing PAGE for analysis of laccases by quantification of the amount of melanin pigment produced by clinical and environmental strains of C. neoformans. After incubation of the gel with the substrate l-dopa, strains produced melanin spots of a bright brown to black colour. Quantification of these spots was performed by densitometry analysis and the amount of melanin produced was calculated and compared among the strains. All strains showed laccase activity. Serotype B strains showed a higher melanin intensity than serotype A strains. Over half of the clinical strains (56.2 %) showed the lowest melanin intensities, suggesting that melanin production may not be the main virulence factor against host defence. The clinical strain ICB 88 revealed two melanin spots on the gel, indicating the presence of two laccase isoforms. The environmental strains showed the highest values of melanin intensity, which may be related to previous exposure to environmental stress conditions.

This study was conducted to evaluate the effects of the ethanolic extract of Passiflora edulis leaves on blood glucose, protein glycation, NADPH oxidase activity and macrophage phagocytic capacity after Candida albicans exposure in diabetic rats. The Passiflora edulis Sims leaves were dried to 40°C, powdered, extracted by maceration in 70% ethanol, evaporated under reduced pressure and lyophilised. The biochemical tests performed were total phenolic content (TP) as determined by the Folin-Ciocalteu assay, trapping potential DPPH assay and total iron-reducing potential. Diabetes was induced by alloxan injection. Protein glycation was determined by AGE and fructosamine serum concentrations. Extract-treated diabetic animals demonstrated lower fructosamine concentrations compared with the diabetic group. Our results suggest that ethanolic Passiflora edulis Sims leaf extraction may have beneficial effects on diabetes and may improve glycaemic control in diabetic rats. resumo O objetivo deste estudo foi avaliar os efeitos do extrato etanólico de folhas de Passiflora edulis sobre os níveis de glicose sanguínea, glicação protéica, produção de espécies reativas de oxigênio (ERO) e capacidade fagocítica de macrófagos de ratos diabéticos. As folhas de Passiflora edulis Sims foram secas a 40 °C, trituradas e o extrato preparado por maceração em solução hidroetanólica 70% (v/v) etanol foi evaporado sob pressão reduzida e liofilizado. Os testes químicos realizados demonstraram que além da presença de compostos fenólicos, determinada pelo método de Folin-Ciocalteu, o extrato apresentou potencial sequestrante de radicais DPPH e redutor de ferro. Nos animais diabéticos foi observado aumento na glicação protéica, avaliada pela concentração de frutosaminas e de produtos de glicação avançada (AGE), e redução na produção de ERO por macrófagos frente à Candida albicans, quando comparados ao grupo controle. O tratamento dos animais diabéticos com o extrato reduziu as concentrações de frutosaminas e manteve a produção de ERO em níveis semelhantes aos observados no grupo controle. Nossos resultados sugerem que o extrato etanólico de folhas de Passiflora edulis Sims pode apresentar efeitos benéficos sobre o diabetes e melhorar o controle glicêmico em ratos diabéticos.

Polymorphonuclear neutrophils (PMN) participate in an active way in the innate immunity developed after the fungal infection paracoccidioidomycosis (PCM). Nevertheless, the sole participation of neutrophils is not sufficient to eradicate PCM`s pathogenic fungus: Paracoccidioides brasiliensis (Pb). In that way, we aimed to develop a treatment capable of stimulating PMN to the site of injury through low-level laser therapy (LLLT). (LLLT) is safe to use and has not been linked to microorganism resistance so far; in addition, based on previous studies we understand that LLLT may be useful to treat several medical conditions through the stimulation and activation of certain types of cells. This brief review is based on the novel attempt of activating PMN against a fungal infection.

A. crassiflora Mart. a Brazilian savannah fruit, is a source of phytochemical compounds that possess a wide array of biological activities, including free radical scavenging. This native fruit proved to potentialize the mutagenic process in previous in vivo investigations. The aim of the present study was to investigate the effects of A. crassiflora Mart. pulp intake on colonic cell proliferation and on the development of Aberrant Crypt Foci (ACF) in male Wistar rats. The animals were fed with either a commercial diet or a diet supplemented with A. crassiflora Mart. pulp mixed in 1%, 10% or 20% (w/w) for 4 weeks or 20 weeks. The carcinogen 1,2-dimethylhydrazine dihydrochloride (4 doses, 40 mg kg-1 each) was used to induce colonic ACF. After euthanasia, the blood, liver and colon samples were collected for biochemical determinations, oxidative stress or ACF development analysis, respectively. Immunohistochemical analyses of the colonic mucosa were performed using antibodies against proliferating cell nuclear antigen (PCNA) in normal-appearing colonic crypt and β-catenin in ACF. There was no ACF development in the colon from groups treated with A. crassiflora Mart. pulp. Also, the biochemical and oxidative stress analysis, PCNA labeling and ACF development (number, multiplicity or cellular localization of β-catenin) were unchanged as a result of marolo pulp intake. Thus, the present results suggest that A. crassiflora Mart. pulp intake did not exert any protective effect in the colon carcinogenesis induced by DMH in rats.

Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) and other cyclic nitroxides have been shown to inhibit the chlorinating activity of myeloperoxidase (MPO) in vitro and in cells. To examine whether nitroxides inhibit MPO activity in vivo we selected acute carrageenan-induced inflammation on the rat paw as a model. Tempol and three more hydrophobic 4-substituted derivatives (4-azido, 4-benzenesulfonyl, and 4-(4-phenyl-1H-1,2,3-triazol-1-yl)) were synthesized, and their ability to inhibit the in vitro chlorinating activity of MPO and carrageenan-induced inflammation in rat paws was evaluated. All of the tested nitroxides inhibited the chlorinating activity of MPO in vitro with similar IC50 values (between 1.5 and 1.8μM). In vivo, the attenuation of carrageenan-induced inflammation showed some correlation with the lipophilicity of the nitroxide at early time points but the differences in the effects were small (200-fold). No inhibition of MPO activity in vivo was evident because the levels of MPO activity in rat paws correlated with the levels of MPO protein. Likewise, paw edema, levels of nitrated and oxidized proteins, and levels of plasma exudation correlated with the levels of MPO protein in the paws of the animals that were untreated or treated with the nitroxides. The effects of the nitroxides in vivo were compared with those of 4-aminobenzoic hydrazide and of colchicine. Taken together, the results indicate that nitroxides attenuate carrageenan-induced inflammation mainly by reducing neutrophil migration and the resulting MPO-mediated damage. Accordingly, tempol was shown to inhibit rat neutrophil migration in vitro.

In this work the polyphenol oxidase (PPO) was the main component of a biosensor for adrenaline determination. The activity of this enzyme was measured in several vegetables. Banana (Musa sp.) extracts presented better results with 974 UA (units of activity). The biosensor was constructed with a polyethylene tube (0.8 mm i.d.) filled with: carbon paste containing 50 UA of the PPO in phosphate buffer (pH=7.00) solution and vaseline as agglutinant. When the biosensor was applied in medicine samples it provided a linear range from 8.00×10(-9) to 8.00×10(-4) mol L(-1); the results obtained with the proposed method and the Brazilian Pharmacopoeia method were in agreement (t-test). When it was applied in blood samples, the matrix-matching calibration was used, and the linear range was from 8.00×10(-7) to 8.00×10(-3) mol L(-1). In vivo studies were also done. The obtained results for those electrodes, which were inserted in the jugular vein of Wistar rats, were very promising.

A new method for saccharin determination in liquid sweetener products was developed. The method is based on the precipitation reaction of Ag(I) ions with saccharin in aqueous medium (pH 3.0), using a flow injection analysis system with merging zones, the suspension was stabilized with 5 g L−1 Triton X-100. All experimental parameters influencing the flow injection system were optimized by means of chemometric approaches. The linear analytical curve was built from 2.4 g L−1 up to 9.64 g L−1 (r = 0.9968) with a quantification limit of 2.40 g L−1. The precision assessed as relative standard deviation (n = 10) was found to be 1.75 % for the saccharin concentration of 7.20 g L−1. Based on interference studies performed with the substances commonly found in liquid sweeteners, such as sodium cyclamate, methylparaben, sodium aspartame, and benzoic and citric acids, at the analyte to interferent mole ratio of up to 1: 10, no interference with the saccharin determination was observed. The presence of chloride ions interferes with the method, but a preceding liquid-liquid saccharin extraction with ethyl acetate was successfully employed to overcome this drawback. Accuracy of the method in sweetener products was evaluated by a comparison with the HPLC method.

The crystalline structure of mangiferin (=2-β-D-glucopyranosyl-1,3,6,7-tetrahydroxy-9H-xanthen-9-one; 1), a biologically active xanthenone C-glycoside, isolated from the stem bark of Mangifera indica (Anacardiaceae), was unambiguously determined by single-crystal X-ray diffraction (XRD). The crystal structure is summarized as follows: triclinic, P1, a=7.6575(5), b=11.2094(8), c=11.8749(8) A, α=79.967(5), β=87.988(4), γ=72.164(4)°, V=955.3(1) A3, and Z=2. The structure also shows two molecules in the asymmetric unit cell and five crystallization H2O molecules. The packing is stabilized by several intermolecular H-bonds involving either the two symmetry-independent mangiferin molecules 1a and 1b, or the H2O ones.

Neutrophils (PMN) play a central role in host defense against the neglected fungal infection paracoccidioidomycosis (PCM), which is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb). PCM is of major importance, especially in Latin America, and its treatment relies on the use of antifungal drugs. However, the course of treatment is lengthy, leading to side effects and even development of fungal resistance. The goal of the study was to use low-level laser therapy (LLLT) to stimulate PMN to fight Pb in vivo. Swiss mice with subcutaneous air pouches were inoculated with a virulent strain of Pb or fungal cell wall components (Zymosan), and then received LLLT (780 nm; 50 mW; 12.5 J/cm2; 30 seconds per point, giving a total energy of 0.5 J per point) on alternate days at two points on each hind leg. The aim was to reach the bone marrow in the femur with light. Non-irradiated animals were used as controls. The number and viability of the PMN that migrated to the inoculation site was assessed, as well as their ability to synthesize proteins, produce reactive oxygen species (ROS) and their fungicidal activity. The highly pure PMN populations obtained after 10 days of infection were also subsequently cultured in the presence of Pb for trials of protein production, evaluation of mitochondrial activity, ROS production and quantification of viable fungi growth. PMN from mice that received LLLT were more active metabolically, had higher fungicidal activity against Pb in vivo and also in vitro. The kinetics of neutrophil protein production also correlated with a more activated state. LLLT may be a safe and non-invasive approach to deal with PCM infection., Conselho Nacional de Pesquisas (Brazil) (486135/2012-8), Conselho Nacional de Pesquisas (Brazil) (304827/2012-6), Fundacao de Amparo a Pesquisa do Estado de Minas Gerais (CBB-PPM-00119-14), National Institutes of Health (U.S.) (R01AI050875)

Foi estudado o efeito da lactona sesquiterpênica licnofolido sobre o \"burst\" respiratório de leucócitos polimorfonucleares inflamatórios (PMN) estimulados por forbol (PMA), pelo peptídeo quimiotático fMLP ou zimozan opsonizado (OZ). O licnofolido inibiu de forma dose-dependente a liberação de O2•- pelos PMN, sem alteração do período \"Iag\" do complexo NADPH. oxidase. O efeito foi mais acentuado quando os PMN foram estimulados diretamente pela via de proteína quinase C. A adição de ditiotreitol ou glutationa reduzida (GSH) às suspensões celulares antes da incubação com licnofolido preveniu parcialmente o efeito inibitório. O tratamento dos PMN com a lactona determinou uma queda drástica dos níveis celulares de GSH livre, sem incremento de glutationa oxidada (GSSG). A reação direta entre GSH e licnofolido foi confirmada com a detecção de um aduto glutationil-licnofolido através de identificação por espectrometria de massa (ESI-MS/MS). A S-tiolação protéica induzida pelo PMA foi reduzida em PMN tratados com Iicnofo/ido, como detectado através de determinação de incorporação de [35S], sendo que 80% desses tióis foram identificados como GSH. Uma série de proteínas S-glutationadas foi detectada através de autoradiografias, sendo que aquelas correspondentes a 38 e 24 kDa tiveram essa modificação póstraducional suprimida pelo tratamento com dose de licnofolido capaz de suprimir o \"burst\" respiratório dos PMN. Estes resultados indicam que a depleção celular de GSH causada pelo licnofolido impede a sustentação do \"burst\" respiratório pelos PMN, em correlação direta com a diminuição de S-glutationação protéica. An investigation was made into the action of the sesquiterpene lactone lychnopholide on the respiratory burst of inflammatory polymorphonuclear leukocytes. Lychnopholide determined concentration-related inhibition of the generation of phorbol 12-myristate 13-acetate-, chemotatic peptide-, and opsonized zymozan-induced superoxide anion with no effect on the lag time of the assembly of the NADPH oxidase complex, such action was greater on the protein kinase C pathway that on both membrane receptor dependent stimuli via. Subsequent additions of D-glucose, Ca2+, Mg2+, dithiothreitol ar reduced glutathione (GSH) did not reverse the inhibitory action. The addition of both thiols prior to the lychnopholide treatment partially hindered the inhibition rate. The endogenous level of GSH in leukocytes was drastically depleted under the lychnopholide treatment, without corresponding increases occurring in the oxidized form (GSSG). A direct reaction between glutathione and lychnopholide was confirmed from a glutathionyl-lychnopholide adduct detected by electrospray mass spectrometry analysis and identified by tandem mass analysis in cellular extracts. Protein S-thiolation induced by PMA stimulation was decreased in lactone-treated PMN as detected by [35S] scintillation count, which indicated that about 80% of the thiols were glutathione. A subset of S-glutathionylated proteins was identified through gel electrophoresis, which revealed that the modification of the phorbol-triggered protein sulfhydryl in the protein bands corresponding to 38 and 24 kDa was precluded by the lychnopholide treatment correlated with respiratory burst inhibition. These results show that GSH depletion determined by lychnopholide treatment renders PMN to sustain respiratory burst, whose action is proportional to protein S-glutahionylation decrease.

article i nfo This paper describes the investigation of the cytotoxic and antioxidant activities and in vivo mutagenic/ antimutagenic potential of different concentrations of the hexane extract (EHP) and isolated molecule 7-epi-clusianone (MI) of Rheedia brasiliensis. The in vitro antioxidant activity of MI was investigated by mon- itoring the reduction of radical scavenging and metal chelating activity of DPPH (1,1-diphenyl-2-picryl- hydrazil). Cytotoxic activity was assessed by measuring the mortality of brine shrimp in the presence and absence of the compounds. The mutagenic, antimutagenic and cytotoxic effects of these compounds were evaluated by a micronucleus test. During the antioxidant activity assessment, the 7-epi-clusianone was significantly higher than that of EHP at all concentrations in three assays. From the results obtained with the assessment of cytotoxic activity, all samples had a mortality rate (LC50b100 mg/mL) lower than the pos- itive control (thymol). The results of the micronucleus test revealed that MI at 5, 10 and 15 mg/kg b.w. is antimutagenic. In conclusion, these results suggest that in the future, the EHP and MI could be used as prophylactic agents in cancer prevention.

A planta Psidium guajava Linn. (Myrtaceae), popularmente conhecida por goiabeira, é muito utilizada na medicina tradicional devido às suas propriedades antioxidante, antimicrobiana e anti-inflamatória. O objetivo deste trabalho foi a obtenção, padronização e avaliação das atividades biológicas de extratos secos de P. guajava obtidos por spray drying, com ênfase na influência exercida por diferentes adjuvantes tecnológicos sobre as propriedades físicoquímicas dos extratos padronizados. A primeira etapa do trabalho foi a coleta das folhas de quatro cultivares de Psidium guajava (Paluma, Pedro Sato, Século XXI e Santana da Vargem). A variedade Pedro Sato apresentou os maiores teores de substâncias bioativas correlacionadas com a atividade antioxidante, sendo, portanto selecionada para prosseguimento do trabalho. A solução extrativa foi obtida utilizando o método de extração por maceração dinâmica à temperatura de 56 ºC, por 60 min, utilizando etanol 70% (v/v) como solvente extrator e proporção planta/solvente de 1/10 (m/v). O extrato obtido foi concentrado até teor de sólidos de 10,7 % e acrescido dos adjuvantes de secagem nas proporções de 40 e 80% (base seca). Os adjuvantes empregados neste trabalho (isolados ou em associação) foram a goma arábica (Encapsia), amido modificado (Dry Flo), ?- ciclodextrina (Kleptose), celulose microcristalina (Microcel MC 102), dióxido de silício colloidal (Aerosil 200) e maltodextrina (Mor-Rex 1910). A etapa de secagem foi desenvolvida em um spray dryer de bancada modelo SD-05 da LabPlant, sendo os parâmetros operacionais fixados em: temperatura de entrada do ar de secagem (Tge): 150 ºC, vazão do ar de secagem (Wg): 60 m3/h, vazão de alimentação do extrato (Wsusp): 4 g/min, pressão do ar de atomização (Patm): 1,5 bar e vazão do ar de atomização (Watm): 15 lpm. Os extratos secos obtidos foram caracterizados quanto às características físico-químicas, morfológicas e atividade antioxidante. Foram selecionadas três amostras de extratos secos (maltodextrina:Aerosil, maltodextrina:encapsia:aerosil e ?-ciclodextrina na proporção de 80%) para os estudos de estabilidade e ensaios de atividade antioxidante (DPPH, ABTS, ORAC, FRAP e ensaios em modelos celulares, QLlum) e antimicrobiana (bactérias e fungos). A atividade dos extratos secos de P. guajava sobre o metabolismo oxidativo de neutrófilos humanos estimulados com PMA pelo método da quimioluminescência mostra resultados de inibição da QLlum de forma dependente da concentração e não tóxica sobre os neutrófilos. Os extratos secos foram ativos contra S. aureus e C. glabrata, e inativos contra as bactérias Gram negativas avaliadas. Os resultados deste estudo podem contribuir para o desenvolvimento de novas formas farmacêuticas e para a validação de aplicações clínicas da Psidium guajava, espécie inserida no elenco de espécies vegetais selecionadas para estudos pelo Programa de Pesquisas de Plantas Medicinais e incluída na Relação Nacional de Plantas Medicinais de Interesse ao SUS (RENISUS), justificando a importância da investigação da sua viabilidade tecnológica. Psidium guajava Linn. (Myrtaceae), popularly known as guava, is widely used in traditional medicine due to its antioxidant, antimicrobial and anti-inflammatory properties. The aim of this work was to standardization and evaluation of the biological activities of spray drying dried extracts of P. guajava, with emphasis on the influence exerted by different carriers on the physicochemical properties of dried extracts. The first stage of the study was the collection of leaves of four cultivars of Psidium guajava (Paluma, Pedro Sato, Century XXI and Santana\'s Vargem). The variety Pedro Sato showed the highest levels of bioactive substances correlated with antioxidant activity, and therefore selected for further work. The extraction solution was obtained using the dynamic maceration method at a temperature of 56 ° C for 60 min using ethanol 70% (v/v) as extractor solvent and plant/solvent ratio of 1/10 (m/v). The extract obtained was concentrated until a solid contents of 10.7% and added with the carriers in proportions of 40 to 80% (dry basis). The carriers used in this study (alone or in blends) were arabic gum (Encapsia), modified starch (Dry Flo), ?-cyclodextrin (Kleptose), microcrystalline cellulose (MC Microcel 102), colloidal silicon dioxide (Aerosil 200) and maltodextrin (Mor-Rex 1910). The drying runs was performed in a bench top spray dryer model SD-05 LabPlant, and the operational parameters set at: inlet temperature of the drying air (Tgi): 150 °C, flow rate of the drying air (Wg): 60 m3/h, feed flow rate of extract (Wsusp): 4 g/min, atomizing air pressure (Patm): 1.5 bar and atomizing air flow rate (Watm): 15 lpm. Spray dried preparations were characterized by physico-chemical, morphological and antioxidant activity. Three samples of dried extracts was selected (maltodextrin: Aerosil, maltodextrin: encapsia: aerosil and ?-cyclodextrin in the proportion of 80%) for stability studies and trials of antioxidant activity (DPPH, ABTS, ORAC, FRAP and assays in cell models, QLlum) and antimicrobial (bacteria and fungi). The activity of dried extracts of P. guajava by cellular test that measured the luminol-enhanced chemiluminescence (CLlum) produced by phorbol myristate acetate (PMA) stimulated neutrophils show results QLlum inhibition of concentration-dependent manner and nontoxic on neutrophils. The dried extracts were active against S. aureus and C. glabrata, and inactive against Gram-negative bacteria evaluated. The results of this study may contribute to the development of novel dosage forms for validation and clinical applications of the Psidium guajava, medicinal specie included in the list of plant species selected for study by the Research Program on Medicinal Plants and included in the National List of Medicinal Plants Interest SUS (RENISUS), justifying the importance of research of its technological feasibility.