Caffeine improves the cytotoxic effect of dacarbazine on B16F10 murine melanoma cells.

OBJECTIVE: Caffeine has been studied as a potentiating agent in chemotherapy against some types of cancer, but there are few reports on its effects on melanoma. This study aimed to investigate caffeine's ability to enhance the effects of dacarbazine in vitro. MATERIALS AND METHODS: Murine melanoma B16F10 cells were treated 24 h with 1-40 µM caffeine. We evaluated cytotoxicity, DNA damage, apoptosis, and oxidative lesion induced by dacarbazine associated with caffeine. The metabolization of these drugs, as well as immunocytochemical labeling, were also evaluated. CONCLUSIONS: The pre-treatment with caffeine showed to be more effective. Caffeine potentiated dacarbazine-induced cytotoxic effects by increasing dacarbazine biotransformation, apoptosis, DNA damage, and malondialdehyde levels; also, caffeine reduced Ki67 and ERK1/2 nuclear labeling and increased p53 labeling in B16F10 cells. In our experiment, caffeine promoted modifications associated with dacarbazine metabolism by viable cells potentiating this antineoplastic drug. These promising results should be further evaluated in experimental models in vivo.

Adsorbent selection for pesticides removal from drinking water.

Pesticides present in water resources can cause adverse health effects even in low concentrations, due to their bio-accumulative character. Therefore, the challenge for drinking water production increases, due to the limitations of conventional water treatment technologies in the removal of small molecular weight dissolved compounds. This work aimed to provide technical and scientific support for the selection of pulverized activated carbon - PACs, granular activated carbon - GACs, and carbon nanotubes - CNT concerning atrazine - ATZ, simazine - SMZ, and diuron - DIU adsorption for application in water treatment plants, considering two forms of application commercial product - CP and analytical standard - SD. These forms of application were tested aiming to verify the influence of the purity of the products used in experiments on the adsorption efficiency. It was possible to verify the adsorption efficiencies were not guided only by the characteristics of the adsorptive materials used, and that the selection should not be carried out only based on the, specific superficial area - BET size and distribution of specific pore volume. The isotherms demonstrated that the parameter Kf associated with the results of the selection experiment can be considered an alternative technical tool of simple application and sufficient for this purpose. Also, the capacity of activated carbons - ACs and nanomaterials - NMs were affected by the application of the compounds, highlighting the importance of using commercial product - CP in scientific research and technical investigations.Hightlights The pesticides efficiency removal was affected due to the forms of application, SD and CP;The parameters IN and MBI were not decisive in the selection of the activated carbon;The main adsorption mechanism in all the materials was chemical;GAC was the most efficient adsorbent in the removal of the pesticides;An adequate adsorbent selection is crucial for satisfactory removal of pesticides in water.

Chimarrão consumption and prognostic factors in breast cancer: Correlation with antioxidants and blood caffeine levels.

Breast cancer is the second most common malignancy among women. Ilex paraguariensis A. St. Hil, known as yerba mate, is widely consumed in southern Brazil as a hot infusion drink known as chimarrão. This herb has a complex chemical composition and is rich in antioxidants, which may interfere in the course of chronic inflammatory diseases as breast cancer. This study investigated the impact of chimarrão consumption on the clinicopathological profile of women with breast cancer attended at Francisco Beltrão Cancer Hospital, Paraná, Brazil. Blood antioxidants and caffeine profiles were assessed. Decreases in reduced glutathione and metallothionein levels, and increase in catalase activity were observed among breast cancer patients that were chimarrão consumers. The levels of circulating caffeine in breast cancer patients with luminal A tumors were higher than those in patients with luminal B and HER-2 subtypes. Furthermore, overweight patients presented higher caffeine levels than the eutrophic ones. It was found positive associations between chimarrão intake and high body mass index, and chimarrão intake and menopause at diagnosis. Altogether, these findings suggest that chimarrão consumption affects the blood antioxidants of breast cancer patients, and that the caffeine present in this mixture may favor the development of tumor of good prognosis. HIGHLIGHTS: Chimarrão consumption may affect the course of chronic inflammatory diseases, as breast cancer. Chimarrão intake changed blood antioxidants in breast cancer patients who were current consumers when compared to the non-consumers ones. High levels of caffeine were detected in patients bearing luminal A tumors, suggesting a protective role.

Analysis of the phenolic composition and yield of 'BRS Vitoria' seedless table grape under different bunch densities using HPLC-DAD-ESI-MS/MS.

Grapes are considered to be a major source of phenolic compounds as compared with other fruits. To improve the quality of table grapes, some techniques like thinning can be used. In addition, grape cultivars with distinct characteristics are directly linked to its phenolic profile. This study aimed to identify and quantify the phenolic compound profile and yield of the hybrid 'BRS Vitoria' seedless table grape under different bunch densities, using a combination of solid-phase extraction (SPE) methodologies and analytical high-performance liquid chromatography-diode array detector with tandem mass spectrometry (HPLC-DAD-ESI-MS/MS). A trial was carried out in 2016, in a commercial vineyard at Marialva, state of Parana (South Brazil). Three weeks after anthesis, the following bunch densities were evaluated: 4.0, 4.5, 5.0, 5.5, and 6 bunches per m2 (corresponding to an estimation of 16, 18, 20, 22, and 24 tons ha-1). The randomized block design was used as a statistical model with each treatment was replicated four times, with one vine per plot. Different characteristics were evaluated at harvest, e.g., soluble solids content (SS), total acidity (TA), maturation index (MI = SS/TA), bunch and berry masses, yield, as well hydroxycinnamic acid derivative (HCAD), anthocyanin, flavonol, and flavan-3-ol contents by HPLC-DAD-ESI-MS/MS analysis. The evaluated bunch densities did not interfere with the physicochemical characteristics of the berries, such as SS and MI. Under the density of 6.0 bunches per m2, the highest yield of 25 tons ha-1 was reached. Under all bunch densities, the phenolic profile presented the same compounds, but at different concentrations. Under a density of 5.0 bunches per m2, the compounds belonging to the anthocyanin and flavonol families were present in high concentrations. In contrast, at the densities of 4.0 and 4.5 bunches per m2, there was a reduction in the flavan-3-ol content. With respect to stilbenes, only the trans-piceid and its cis- isomer were detected. However, their concentrations had no significant influence on the evaluated bunch densities.

Pretreatment Efficiency Using Autoclave High-Pressure Steam and Ultrasonication in Sugar Production from Liquid Hydrolysates and Access to the Residual Solid Fractions of Wheat Bran and Oat Hulls.

The objective of this study was to evaluate the combination of physical and chemical pretreatments of wheat bran (WB) and oat hulls (OH) to obtain fermentable sugars and a residual solid fraction with increased susceptibility to enzymatic hydrolysis. High-pressure steam in an autoclave and ultrasonication were employed as pretreatments, and for both processes, WB and OH were treated with sulfuric acid (H2SO4), neutral medium (H2O) and sodium hydroxide (NaOH). Autoclave high-pressure steam in an acid medium was the most effective for the release of sugars (total sugars, xylose and glucose) from liquid hydrolysates and for the modification of the residual solid fraction. The cellulose content of the WB residual solid fraction increased from 7.19 to 39.17%, the lignin fraction of WB decreased from 6.40 to 3.21%, the cellulose content of OH increased from 31.16 to 61.53%, and lignin fraction of OH decreased from 18.12 to 7.24%, resulting in materials more susceptible to enzymatic hydrolysis.

Milk with Juçara (Euterpe edulis Martius) Pulp: Fermentation by L. reuteri LR92 and Reuterin Production in Situ

Abstract This study aimed to develop and evaluate fermented milk by Lactobacillus reuteri LR92 with addition of juçara pulp (FMJ) and reuterin production in situ. The fermentation process was analyzed for 24 hours and the storage of FMJ for 30 days at 4 °C. During the fermentation, there was consumption of 25% (w / v) of lactose and increase of 0.01 to 0.85% (w / v) of lactic acid. The FMJ presented 0.43 ± 0.01 mM of reuterin, inhibiting Staphylococcus aureus strains under in vitro test. For the carbohydrates, the percentages (g.100g-1) found were 7.31 ± 1.07; 9.19 ± 0.82; 1.60 ± 0.50 and 0.08 ± 0.00 for sucrose, lactose, galactose and fructose respectively. The survival of L. reuteri, present in FMJ, was 2.47 log CFU / mL after 6 hours of gastrointestinal simulation. In sensory analysis FMJ received a grade 7 for global acceptance indicating good acceptance of the product.

Optimization and validation of an SBSE-HPLC-FD method using laboratory-made stir bars for fluoxetine determination in human plasma.

Depression is the largest cause of disability worldwide, affecting 350 million people. Notwithstanding that clinical trials demonstrate antidepressants efficacy, the efficient response can vary individually concerning therapeutic dosage. Although important, plasma levels monitoring remains an analytical challenge whereas clean-up and pre-concentration represent critical steps. Therefore, this study aims to develop, optimize and validate a method for fluoxetine determination in human plasma, employing a laboratory-made device consisting of a PDMS stir bar sorptive for extraction, coupled with high-performance liquid chromatography-fluorescence detection (SBSE-HPLC-FD). Optimization involved sorption-desorption steps. For sorption, temperature and time were assessed by factorial and central composite design approaches, taking into account the desirability and the response surface results, with stirring speed also examined. For desorption kinetics and ultrasonic and magnetic stirring mode were evaluated. The proposed method after validation was robust, linear (25.00-1000.00 ng mL-1 , R2 > 0.98) and presented good intra- (RSD 4.18%) and inter-day-assay (RSD 11.60%) precision and accuracy (recovery 109.60%), allowing reliable quantitation without interference. The method was successfully applied to real samples. SBSE-HPLC-FD could represent a feasible alternative with good cost-benefit for low-volume samples and therapeutic drug monitoring, as well as contributing to correlation studies between plasma fluoxetine levels and clinical response, which is still little studied.

Caryocar coriaceum extracts exert leishmanicidal effect acting in promastigote forms by apoptosis-like mechanism and intracellular amastigotes by Nrf2/HO-1/ferritin dependent response and iron depletion: Leishmanicidal effect of Caryocar coriaceum leaf exracts.

Leishmania (L.) amazonensis is the American Cutaneous Leishmaniasis-causing agents, and the available drugs for this disease present toxicity, low efficiency and difficulty of administration. Plants belong23ing to the Caryocar genus are found in Brazilian Cerrado, where fruits are used as food and in folk medicine, and previous studies showed several biological effects of extracts of this plant. The present work evaluated the leishmanicidal and immunomodulatory activity of ethyl acetate (EAC) and methanol (MET) C. coriaceum leaf extracts EAC and MET showed an antipromastigote effect after 24, 48 and 72 h. The extracts also induced loss of mitochondrial membrane potential, reactive oxygen species (ROS) production, damage to the plasma membrane, and phosphatidylserine exposure on promastigote forms, and most parasites were going through a late apoptosis-like process. The range of concentrations used did not alter the viability of peritoneal macrophages of BALB/c mice; therefore, we observed that the treatment with extracts was able to reduce the infection of this cells. Thereafter, the extracts were able to significantly improve the levels of TNFα, IL-6, MCP-1, and IL-10, and reduced the levels of MDA and ROS without interfering on NO levels released by infected macrophages. In addition, both EAC and MET up-regulated Nrf2/HO-1/Ferritin expression and reduced the labile iron pool in infected macrophages. Based on the data obtained, it is possible to infer that different solvent extracts of the C. coriaceum leaves exert leishmanicidal effect, acting on promastigote forms through apoptosis-like mechanisms and intracellular amastigote forms involving a Nrf2/HO-1 dependent antioxidant response, which culminates in a depletion of available iron for L. amazonensis replication.

Highly specific changes in antioxidant levels and lipid peroxidation in Parkinson's disease and its progression: Disease and staging biomarkers and new drug targets.

There is evidence that immune-inflammatory, stress of reactive oxygen and nitrogen species (IO&NS) processes play a role in the neurodegenerative processes observed in Parkinson's disease (PD). The aim of the present study was to investigate peripheral IO&NS biomarkers in PD. We included 56 healthy individuals and 56 PD patients divided in two groups: early PD stage and late PD stage. Plasma lipid hydroperoxides (LOOH), malondialdehyde (MDA), nitric oxide metabolites (NOx), sulfhydryl (SH) groups, catalase (CAT) activity, superoxide dismutase (SOD) activity, paraoxonase (PON)1 activity, total radical trapping antioxidant parameter (TRAP) and C-reactive protein (CRP) were measured. PD is characterized by increased LOOH, MDA and SOD activity and lowered CAT activity. A combination of five O&NS biomarkers highly significantly predicts PD with a sensitivity of 94.5% and a specificity of 86.8% (i.e., MDA, SOD activity, TRAP, SH-groups and CAT activity). The single best biomarker of PD is MDA, while LOOH and SOD activity are significantly associated with late PD stage, but not early PD stage. Antiparkinson drugs did not affect O&NS biomarkers, but levodopa+carbidopa significantly increased CRP. It is suggested that MDA may serve as a disease biomarker, while LOOH and SOD activity are associated with late PD stage characteristic. New treatments for PD should not only target dopamine but also lipid peroxidation.

Kaurenoic Acid Possesses Leishmanicidal Activity by Triggering a NLRP12/IL-1ß/cNOS/NO Pathway.

Leishmania amazonensis (L. amazonensis) infection can cause severe local and diffuse injuries in humans, a condition clinically known as American cutaneous leishmaniasis (ACL). Currently, the therapeutic approach for ACL is based on Glucantime, which shows high toxicity and poor effectiveness. Therefore, ACL remains a neglected disease with limited options for treatment. Herein, the in vitro antiprotozoal effect and mechanisms of the diterpene kaurenoic acid [ent-kaur-16-en-19-oic acid] (KA) against L. amazonensis were investigated. KA exhibited a direct antileishmanial effect on L. amazonensis promastigotes. Importantly, KA also reduced the intracellular number of amastigote forms and percentage of infected peritoneal macrophages of BALB/c mice. Mechanistically, KA treatment reestablished the production of nitric oxide (NO) in a constitutive NO synthase- (cNOS-) dependent manner, subverting the NO-depleting escape mechanism of L. amazonensis. Furthermore, KA induced increased production of IL-1ß and expression of the inflammasome-activating component NLRP12. These findings demonstrate the leishmanicidal capability of KA against L. amazonensis in macrophage culture by triggering a NLRP12/IL-1ß/cNOS/NO mechanism.

In vitro and in vivo antileishmania activity of sesquiterpene lactone-rich dichloromethane fraction obtained from Tanacetum parthenium (L.) Schultz-Bip.

The discovery of new treatments for neglected diseases, including leishmaniasis, is a substantial challenge for scientific research. Plant extracts have shown potential in the selective treatment of tropical diseases. The present study evaluated the in vitro and in vivo antileishmania effects of a sesquiterpene lactone-rich dichloromethane fraction (DF) obtained from the aerial parts of Tanacetum parthenium (L.) Schultz-Bip. In vitro studies of the DF indicated an IC50 of 2.40±0.76 µg mL(-1) against the promastigote form and 1.76±0.25 µg mL(-1) against the axenic amastigote form of Leishmania amazonensis. In vivo intramuscular treatment with DF decreased the growth and size of footpad lesions in mice. The DF also significantly decreased the parasite population compared with animals that were treated with the reference drug. Plasma malondialdehyde levels were increased slightly by the DF, attributable to its parthenolide-rich composition that causes cellular apoptosis, compared with the control group, demonstrating treatment efficacy without toxicity or genotoxicity. Because the isolation and purification of plant compounds are costly and time-consuming and generate low yields, extract fractions, such as the DF studied herein, represent a promising alternative for the treatment of leishmaniasis.

Aflatoxin M1 in the urine of non-carriers and chronic carriers of hepatitis B virus in Maringa, Brazil

Exposure to aflatoxins (AFs) in the diet may favour the development of hepatocellular carcinoma (HCC) and the acute exacerbation of hepatitis in chronic hepatitis B virus (HBV) carriers. Measurement of biomarkers such as aflatoxin M1 (AFM1), a metabolite of aflatoxin B1 (AFB1), in urine allows for the assessment of populations exposed to aflatoxins. The aim of this study was to investigate the occurrence of aflatoxin M1 in the urine of HBV carrier and non-carrier patients. One group included 43 randomly selected HBV carriers treated at two hospitals in the city of Maringa, Brazil, from March to June 2008. Control group consisted of 29 healthy adult volunteers with anti-HBs positive and HBsAg negative test results. Detection of AFM1 was performed by fluorescence using high performance liquid chromatography (HPLC) and post-column derivation with the Kobra Cell®. Of the 72 samples analysed, 05/29 (17.2%) AFM1 positive samples were from HBV non-carriers, and 16/43 (37.2%) of samples were from chronic HBV carriers. This study showed AFM1 in the urine of the two surveyed population. However, there is evidence that the chronic HBV carriers have a higher risk of developing HCC due to additive interaction between AFs and HBV.

A exposição às aflatoxinas (AFs) na dieta é um fator de risco para o desenvolvimento do carcinoma hepatocelular (CHC) e a exacerbação da hepatite aguda em indivíduos portadores do vírus da hepatite B (VHB). O uso de biomarcadores, como a aflatoxina M1 (AFM1) na urina, produto de biotransformação da aflatoxina B1 (AFB1), permite avaliar se a população está exposta às AFs. O objetivo do presente estudo foi investigar ocorrência de AFM1 na urina de portadores e não portadores crônicos do VHB. Foi selecionado um grupo, de forma aleatória, representado por 43 portadores do VHB atendidos em dois hospitais da cidade de Maringá, Brasil, no período de Março a Junho/2008. O grupo controle foi composto por 29 voluntários adultos saudáveis anti-HBs positivo e HBsAg negativo. A determinação de AFM1 foi realizada por meio de detecção por fluorescência em sistema de cromatografia a líquido de alta eficiência com derivação pós-coluna utilizando Kobra Cell®. Das 72 amostras analisadas, 05/29 (17,2%) foram positivas para AFM1 em indivíduos não portadores do VHB, e 16/43 (37,2%) de pacientes portadores do VHB. Este estudo demonstrou a ocorrência de AFM1 na urina dos dois grupos estudados. Entretanto, há evidências de que os portadores do VHB possuam alto risco no desenvolvimento do CHC devido ao efeito aditivo pela interação entre aflatoxinas e VHB.

Brazilian red wines made from the hybrid grape cultivar Isabel: phenolic composition and antioxidant capacity.

The hybrid grape cultivar Isabel (V. vinifera x V. labrusca) accounts for about 50% of Brazilian grape production. The anthocyanin composition of the Isabel grape is known, but there is no data for the red wines produced with it. By means of HPLC-DAD-ESI-MS(n) we have studied the most important low molecular phenolic compounds in Isabel red table wines (anthocyanins, pyranoanthocyanins, flavonols and hydroxycinnamic acid derivatives) in order to look for differences between these and V. vinifera red wines, and also their antioxidant capacity using the DPPH method. The anthocyanins found in Isabel red wines are mainly based on malvidin and they were the expected 3-glucosides and the non-vinifera characteristic 3,5-diglucosides including several minor compounds, such as the newly reported cis isomer of malvidin 3-(6''-coumaroyl)-glucoside-5-glucoside. As also found in V. vinifera red wines, Isabel red wines contain pyranoanthocyanins formed from anthocyanidin 3-glucosides during alcoholic fermentation and wine aging (vitisins A and B, and hydroxyphenyl-pyranoanthocyanins). The content of malvidin- and peonidin-based hydroxyphenyl-pyranoanthocyanins derived from p-coumaric and caffeic acids accounted importantly for the pool of low molecular red wine pigments in Isabel wines. The occurrence of other non-anthocyanin phenolic compounds, like hydroxycinnamic acid derivatives and flavonols, are newly reported for Isabel red wines. In contrast to the differences found in the anthocyanins profiles, the flavonols and hydroxycinnamic acid derivatives present in Isabel wines were the same as those in V. vinifera red wines. The content of flavonols is relatively low, but the content in hydroxycinnamic acid derivatives is high in Isabel red wines. Finally, Isabel red wines showed low-medium antioxidant capacity when compared to V. vinifera red wines.

Chemometric evaluation of adulteration profile in coffee due to corn and husk by determining carbohydrates using HPAEC-PAD.

The detection of impurities in coffee samples already roasted and ground is a constant concern mainly in order to verify the incidence of frauds. Carbohydrates contents are important as variations on original constitutes of different matrixes may be able to reveal the final composition of the product. In this sense, a study is performed in this paper in order to evaluate the quality through concentration of total carbohydrates in Arabic roasted and ground coffee. Chemometric methods were applied in order to verify an adulteration pattern by coffee husk and corn, by the mixture of different amounts of these contaminants to coffee, following a statistical design of Simplex-Centroid. It could be concluded that this method was efficient in distinguishing different matrixes. In pure coffee, higher concentrations were found for galactose and mannose with levels of 8.25% and 9.65% (w/w), respectively. However, in the pure husks, the main carbohydrates were mannitol (with a level of 0.64%), arabinose (with 4.24%), and xylose (with 3.40%). For the corn sample, glucose was detected in greater quantity, with 52.53% (w/w). Models presenting the influence of adulterants incorporated to coffee in the carbohydrate level were obtained.

Plasma malondialdehyde levels and CXCR4 expression in peripheral blood cells of breast cancer patients.

INTRODUCTION: Breast cancer is one of the most common neoplasms in women and is a leading cause of cancer related deaths worldwide. Chemokines and their receptors are involved in the control of lymphocyte traffic, a critical component of systemic immunity. CXCR4 mRNA could be involved in the development of variety of diseases. Lipid peroxidation, the result of nonenzymatic autooxidation of polyunsaturated fatty acids, presents numerous harmful effects on biological systems and has been implicated in diseases like cancer. This study examined CXCR4 mRNA expression in peripheral blood cells and malondialdehyde (MDA) in plasma from blood donors and breast cancer patients. MATERIALS AND METHODS: CXCR4 expression in peripheral blood cells from 59 breast cancer patients and 76 healthy blood donors was analyzed by real-time PCR. Plasma MDA was analyzed using high-performance liquid chromatography (HPLC). CONCLUSION: In all stages, MDA levels in total breast cancer patients (1.41 +/- 0.11) were significantly higher (P < 0.01) than those in healthy subjects (0.34 +/- 0.03). No statistically significant difference occurred between CXCR4 expression in peripheral blood cells from breast cancer patients (1.69 +/- 1.05) and the normal healthy control group (1.8 +/- 0.65). However, stage II samples differed statistically (4.3 +/- 1.72) from control, total cancer patients and stages I, III and IV samples.

Association between soy and green tea (Camellia sinensis) diminishes hypercholesterolemia and increases total plasma antioxidant potential in dyslipidemic subjects.

OBJECTIVE: To evaluate the hypolipemic and antioxidant effects of soy and green tea alone and/or in association in dyslipidemic subjects. METHODS: One hundred dyslipidemic individuals were allocated into four groups. The soy group ingested 50 g of soy (kinako) daily, and the green tea group ingested 3 g of green tea in 500 mL of water per day. A third group ingested 50 g of soy and 3 g of green tea daily, and the control group had a hypocholesterolemic diet. Evaluations were performed at baseline and after 45 and 90 d. Plasma levels of total cholesterol, high-density lipoprotein, and triacylglycerols were evaluated by automated methods. Low-density lipoprotein (LDL) cholesterol was calculated using the Friedewald equation. LDL was isolated by ultracentrifugation. Total plasma antioxidant capacity and plasma levels of total lipid hydroperoxides and those linked to LDL were evaluated by chemiluminescence. The results were expressed as median values and their 25th to 75th percentiles, with a 5% level of significance. RESULTS: No significant difference occurred in LDL, high-density lipoprotein cholesterol, and triacylglycerol levels across groups. However, a statistically significant difference in total cholesterol occurred within the soy/green tea group 45 and 90 d after intervention. No statistically significant difference occurred in plasma levels of lipid hydroperoxides or those linked to LDL in any of the groups studied. All the groups that used soy and/or green tea presented increased total plasma antioxidant potential. CONCLUSION: Soy and green tea, alone or in combination, increased the total antioxidant potential of hypercholesterolemic patients, whereas only the combination decreased total cholesterol levels.