In vitro and in vivo antitumor activity of crude extracts obtained from Brazilian Chromobacterium sp isolates

Natural products produced by microorganisms have been an important source of new substances and lead compounds for the pharmaceutical industry. Chromobacterium violaceum is a Gram-negative β-proteobacterium, abundant in water and soil in tropical and subtropical regions and it produces violacein, a pigment that has shown great pharmaceutical potential. Crude extracts of five Brazilian isolates of Chromobacterium sp (0.25, 2.5, 25, and 250 µg/mL) were evaluated in an in vitro antitumor activity assay with nine human tumor cells. Secondary metabolic profiles were analyzed by liquid chromatography and electrospray ionization mass spectrometry resulting in the identification of violacein in all extracts, whereas FK228 was detected only in EtCE 308 and EtCE 592 extracts. AcCE and EtCE 310 extracts showed selectivity for NCI/ADR-RES cells in the in vitro assay and were evaluated in vivo in the solid Ehrlich tumor model, resulting in 50.3 and 54.6% growth inhibition, respectively. The crude extracts of Chromobacterium sp isolates showed potential and selective antitumor activities for certain human tumor cells, making them a potential source of lead compounds. Furthermore, the results suggest that other compounds, in addition to violacein, deoxyviolacein and FK228, may be involved in the antitumor effect observed.

Role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism

Our aim was to investigate the role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism. Coronary angiography and intravascular ultrasound (IVUS) were performed in 60 stable angina pectoris (SAP) patients and 60 unstable angina pectoris (UAP) patients. The chemotactic activity of monocytes in the 2 groups of patients was examined in Transwell chambers. High-sensitivity C-reactive protein (hs-CRP), monocyte chemoattractant protein-1 (MCP-1), regulated on activation in normal T-cell expressed and secreted (RANTES), and fractalkine in serum were examined with ELISA kits, and expression of MCP-1, RANTES, and fractalkine mRNA was examined with real-time PCR. In the SAP group, 92 plaques were detected with IVUS. In the UAP group, 96 plaques were detected with IVUS. The plaques in the UAP group were mainly lipid 51.04% (49/96) and the plaques in the SAP group were mainly fibrous 52.17% (48/92). Compared with the SAP group, the plaque burden and vascular remodeling index in the UAP group were significantly greater than in the SAP group (P<0.01). Chemotactic activity and the number of mobile monocytes in the UAP group were significantly greater than in the SAP group (P<0.01). Concentrations of hs-CRP, MCP-1, RANTES, and fractalkine in the serum of the UAP group were significantly higher than in the serum of the SAP group (P<0.05 or P<0.01), and expression of MCP-1, RANTES, and fractalkine mRNA was significantly higher than in the SAP group (P<0.05). MCP-1, RANTES, and fractalkine probably promote instability of coronary atherosclerotic plaque.

Specific alterations in plasma proteins during depressed, manic, and euthymic states of bipolar disorder

Bipolar disorder (BD) is a common psychiatric mood disorder affecting more than 1-2% of the general population of different European countries. Unfortunately, there is no objective laboratory-based test to aid BD diagnosis or monitor its progression, and little is known about the molecular basis of BD. Here, we performed a comparative proteomic study to identify differentially expressed plasma proteins in various BD mood states (depressed BD, manic BD, and euthymic BD) relative to healthy controls. A total of 10 euthymic BD, 20 depressed BD, 15 manic BD, and 20 demographically matched healthy control subjects were recruited. Seven high-abundance proteins were immunodepleted in plasma samples from the 4 experimental groups, which were then subjected to proteome-wide expression profiling by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-time-of-flight/time-of-flight tandem mass spectrometry. Proteomic results were validated by immunoblotting and bioinformatically analyzed using MetaCore. From a total of 32 proteins identified with 1.5-fold changes in expression compared with healthy controls, 16 proteins were perturbed in BD independent of mood state, while 16 proteins were specifically associated with particular BD mood states. Two mood-independent differential proteins, apolipoprotein (Apo) A1 and Apo L1, suggest that BD pathophysiology may be associated with early perturbations in lipid metabolism. Moreover, down-regulation of one mood-dependent protein, carbonic anhydrase 1 (CA-1), suggests it may be involved in the pathophysiology of depressive episodes in BD. Thus, BD pathophysiology may be associated with early perturbations in lipid metabolism that are independent of mood state, while CA-1 may be involved in the pathophysiology of depressive episodes.

Effect of (+)-dehydrofukinone on GABAA receptors and stress response in fish model

(+)-Dehydrofukinone (DHF) is a major component of the essential oil of Nectandra grandiflora (Lauraceae), and exerts a depressant effect on the central nervous system of fish. However, the neuronal mechanism underlying DHF action remains unknown. This study aimed to investigate the action of DHF on GABAA receptors using a silver catfish (Rhamdia quelen) model. Additionally, we investigated the effect of DHF exposure on stress-induced cortisol modulation. Chemical identification was performed using gas chromatography-mass spectrometry and purity was evaluated using gas chromatography with a flame ionization detector. To an aquarium, we applied between 2.5 and 50 mg/L DHF diluted in ethanol, in combination with 42.7 mg/L diazepam. DHF within the range of 10-20 mg/L acted collaboratively in combination with diazepam, but the sedative action of DHF was reversed by 3 mg/L flumazenil. Additionally, fish exposed for 24 h to 2.5-20 mg/L DHF showed no side effects and there was sustained sedation during the first 12 h of drug exposure with 10-20 mg/L DHF. DHF pretreatment did not increase plasma cortisol levels in fish subjected to a stress protocol. Moreover, the stress-induced cortisol peak was absent following pretreatment with 20 mg/L DHF. DHF proved to be a relatively safe sedative or anesthetic, which interacts with GABAergic and cortisol pathways in fish.

SURVEY OF DEOXYNIVALENOL, DIACETOXYSCIRPENOL, AND T2 TOXIN IN POPCORN HYBRIDS PLANTED IN THE STATE OF SÃO PAULO AND IN POPCORN COMMERCIALIZED IN THE CITY OF CAMPINAS, SP

The incidence of the trichothecenes deoxynivalenol (DON), diacetoxyscirpenol (DAS), and T2 toxin (T2) in popcorn was investigated in 90 samples, belonging to 2 commercial and 28 experimental hybrids planted at experimental stations of the Agronomic Institute of Campinas at the locations of Campinas, Mococa, and Capão Bonito, and in 15 samples of popcorn, 9 branded and 6 unbranded, acquired from commercial outlets in the city of Campinas, SP. The samples were analyzed by gas chromatography with flame ionization detector combined with an alumina:carbon cleanup column. The detection limits were 30ng/g for DON, 50ng/g for DAS, and 40ng/g for T2. Five samples were contaminated with DON, four of them commercial and one from an experimental cultivar. The level of contamination in the commercial samples ranged from 30 to 40ng/g. The sample from the experimental cultivar contained 770ng/g DON. DAS and T2 were not detected in any of the popcorn samples analyzed.

Postharvest respiratory activity and changes in some chemical constituents during maturation of yellow mombin (Spondias mombin) fruit

Mature fruit from the yellow mombin (Spondias mombin) was monitored for its respiration activity. Mature green fruit from the yellow mombin was stored in closed glass chambers and the concentration of oxygen and carbon dioxide at the end of a six hour respiration period was determined. At the same interval of time, the lid of the chamber was opened for air renewal. The increase in carbon dioxide and decrease in oxygen concentration demonstrated that the fruit was climacteric. The maximum liberation of CO2 54.2 mL Kg-1 h-1 and maximum absorption of O2 49.0 mL Kg-1 h-1 occurred 186 hours after the harvest which, obviously, represented the optimum fruit quality after the senescence process started. The respiratory quotient of fruit at a climacteric maximum was 1.11 representing the oxidation of carbohydrates. Total soluble solids increased from 9.1 °Brix (initial) to 13.7 °Brix (climacteric maximum) during maturation, while the total number of acids in the fruit decreased during maturation i.e. from 1.55% initially to 1.40% at pre-climacteric, 1.0% at climacteric maximum and 0.8% in the post-climacteric stage. A similar behaviour was observed in the case of ascorbic acid. There was a continuous decrease in chlorophyll and a continuous increase in the carotenoid content of fruit during maturation.

A method to determine volatile contaminants in polyethylene terephthalate (PET) packages by HDC-GC-FID and its application to post-consumer materials

A simple and low cost method to determine volatile contaminants in post-consumer recycled PET flakes was developed and validated by Headspace Dynamic Concentration and Gas Chromatography-Flame Ionization Detection (HDC-GC-FID). The analytical parameters evaluated by using surrogates include: correlation coefficient, detection limit, quantification limit, accuracy, intra-assay precision, and inter-assay precision. In order to compare the efficiency of the proposed method to recognized automated techniques, post-consumer PET packaging samples collected in Brazil were used. GC-MS was used to confirm the identity of the substances identified in the PET packaging. Some of the identified contaminants were estimated in the post-consumer material at concentrations higher than 220 ng.g-1. The findings in this work corroborate data available in the scientific literature pointing out the suitability of the proposed analytical method.

Color degradation kinetics in low-calorie strawberry and guava jellies

The purpose of this study was to follow-up color changes in low-calorie strawberry and guava jellies during storage. To this end, one formulation of each flavor was prepared varying the application of hydrocolloids (pectin and modified starch). The jellies were studied regarding pH, soluble solids, water activity and syneresis. In order to follow-up color changes, the samples remained stored for 180 days in chambers with controlled temperatures of 10 °C (control) and 25 °C (commercial), and color instrumental analyses (L*, a*, and b*) were performed every 30 days. Arrhenius model was applied to reaction speeds (k) at different temperatures, where light strawberry and guava jellies showed greater color changes when stored at 25 °C compared to the samples stored at 10 °C. Activation energy values between 13 and 15 kcal.mol-1 and Q10 values between 2.1 and 2.3 were obtained for light strawberry jelly and light guava jelly, respectively. Therefore, it was concluded that, with respect to color changes, every 10 °C temperature increase reduces light jellies shelf-life by half.

Fatty acids profile of pulp and nuts of Brazilian fruits

Fruits and nuts from the North and Northeast regions of Brazil were collected to determine the fatty acid profile of their oils. The species studied were Brazil (Bertholletia excelsa H.B.K.), Mucajá (Couma rigida M.), Inajá (Maximiliana maripa D.), Jenipapo (Genipa Americana L.), and Buriti (Mauritia flexuosa L.) nuts. Fatty acid methyl esters were analyzed by gas chromatography with flame ionization detection (GC-FID). Brazil nut major fatty acid was 18:3n-3 (α-linolenic acid), and Buriti nut had approximately 23 times more 18:3n-3 than the pulp. Mucajá nut presented high content of 12:0 (lauric acid) and 16:0 (palmitic acid), and Mucajá pulp showed significant levels of 18:2n-6 (linoleic acid). Considering the PUFA (polyunsaturated fatty acid) sum values, almost all fruits and nuts analyzed presented very high levels of these compounds. Regarding n-6/n-3 ratio, only Brazil Nut, Buriti Nut, Inajá pulp, and Jenipapo pulp corresponded to the desired profile. These Brazilian fruits and nuts could be of potential interest due to their high nutritive value and lipid content.

Reduction of carcass weight loss in swines

The carcass fast freezing is one of the aspects of great prominence to the final quality of pork. In order to reduce weight loss, two experiments were performed, in which the carcasses were monitored during 20 hours to evaluate the main variables involved during two different freezing processes (standard and proposed) as follows: microbiological quality, storage temperature, relative humidity (RH) and air velocity. In experiment I, the carcasses were submitted to a system using heat shock (2 hours in static tunnel at - 25 °C) and subsequently sent to the equalization chamber. In experiment II, the carcasses were submitted to the heat shock and stored in a chamber with RH between 80-85%. The chambers used in both experiments showed no change in the variables studied (internal temperature of 5 °C and air velocity of approximately 0.3 m/s). However, the relative humidity in the three chambers was evaluated and significant differences were found; as a consequence, high levels of weight loss were observed in both chambers In experiment II there was an increase of RH, which reduced the weight loss of the carcasses.

Encapsulated specialty oils commercialized in São Paulo state, Brazil: evaluation of identity (fatty acid profile) and compliance of fatty acids and Vitamin E contents with nutrition labeling

Encapsulated specialty oils commercialized in São Paulo state, Brazil, were evaluated for their identity (fatty acids profile) and compliance with nutrition labeling (fatty acids and Vitamin E (alpha tocopherol) contents). Twenty one samples [flaxseed oil (6), evening primrose (5), safflower (8), borage (1), and black currant (1)] purchased from local markets or collected by the health surveillance agency were analyzed. The fatty acids and vitamin E contents were analyzed by gas chromatography with flame ionization detector and liquid chromatography with UV detector, respectively. Nine samples were adulterated (5 samples of safflower oil, 3 of flaxseed oil, and one of evening primrose). Among them, 3 flaxseed and 2 safflower oil samples were probably adulterated by the addition of soybean oil. Conjugated linoleic acid (CLA) was found in two safflower oils samples although the sale of oils with conjugated linoleic acid (CLA) is not permitted by the National Health Surveillance Agency in Brazil (ANVISA). Only two samples presented all values in compliance with nutrition labeling (one safflower oil sample and one borage oil sample). The results show that a continuous monitoring of encapsulated specialty oils commercialized in Brazil is necessary including a greater number of samples and sanitary surveillance.

Development of a rapid method for the determination of antibiotic residues in honey using UPLC-ESI-MS/MS

Abstract An accurate, reliable and fast multianalyte/multiclass ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed and validated for the simultaneous analysis of 23 pharmaceuticals, belonging to different classes amphenicols, sulfonamides, tetracyclines, in honey samples. The method developed consists of ultrasonic extraction followed by UPLC–ESI–MS/MS with electrospray ionization in both positive mode and negative mode. The influence of the extraction solvents and mobile phase composition on the sensitivity of the method, and the optimum conditions for sample weight and extraction temperature in terms of analyte recovery were extensively studied. The identification of antibiotics is fulfilled by simultaneous use of chromatographic separation using an Acquity BEH C18 (100 mm x 2.1 mm, 1.7 µm) analytical column with a gradient elution of mobile phases and tandem mass spectrometry with an electrospray ionization. Finally, the method developed was applied to the determination of target analytes in honey samples obtained from the local markets and several beekeepers in Muğla, Turkey. Ultrasonic-extraction of pharmaceuticals from honey samples is a well-established technique by UPLC–ESI–MS/MS, the uniqueness of this study lies in the simultaneous determination of a remarkable number of compounds belonging to 23 drug at the sub-nanogram per kilogram level.