Low-intensity red and infrared laser effects at high fluences on Escherichia coli cultures

Semiconductor laser devices are readily available and practical radiation sources providing wavelength tenability and high monochromaticity. Low-intensity red and near-infrared lasers are considered safe for use in clinical applications. However, adverse effects can occur via free radical generation, and the biological effects of these lasers from unusually high fluences or high doses have not yet been evaluated. Here, we evaluated the survival, filamentation induction and morphology of Escherichia coli cells deficient in repair of oxidative DNA lesions when exposed to low-intensity red and infrared lasers at unusually high fluences. Cultures of wild-type (AB1157), endonuclease III-deficient (JW1625-1), and endonuclease IV-deficient (JW2146-1) E. coli, in exponential and stationary growth phases, were exposed to red and infrared lasers (0, 250, 500, and 1000 J/cm2) to evaluate their survival rates, filamentation phenotype induction and cell morphologies. The results showed that low-intensity red and infrared lasers at high fluences are lethal, induce a filamentation phenotype, and alter the morphology of the E. coli cells. Low-intensity red and infrared lasers have potential to induce adverse effects on cells, whether used at unusually high fluences, or at high doses. Hence, there is a need to reinforce the importance of accurate dosimetry in therapeutic protocols.

CROMATOGRAFIA DE AFINIDADE COM CORANTE RED A versus TROCA IÔNICA-PERMEABILIDADE EM GEL: COMPARAÇÃO DA PRATICIDADE NA PURIFICAÇÃO DE ENTEROTOXINA ESTAFILOCÓCICA A

O presente trabalho compara processos de purificação de enterotoxina estafilocócica A, utilizando cromatografia de afinidade com corante Red A em relação a troca iônica (SP - Sephadex C-25) - permeabilidade em gel (Sephadex G-75). Aplicou-se nas colunas o sobrenadante da cultura de Staphylococcus aureus 722 em caldo contendo 3% de triptona e suplementado com 1% de extrato de levedura, previamente concentradas com Amberlite CG-50. O processo capturou rapidamente a EEA, porém a proporção de 15 mg de resina para 150 mg de toxina causou saturação, recuperando apenas 10 a 30% de toxina do sobrenadante. A cromatografia de afinidade com Red A permitiu a recuperação de 60,87% de toxina aplicada em 76 horas, em relação a 114 horas requeridas para purificação utilizando coluna de troca iônica e permeabilidade em gel, com rendimento de 6,5%. O perfil eletroforético das amostras purificadas indicaram que, a toxina obtida da coluna Red A apresentou teor de pureza superior, na ordem de 90%, em relação a 60% atingida pelo método clássico.

Photon Upconverting Nanophosphors: Unique Reporters for Optical Biosensing

Upconversion photoluminescence is a unique property of mostly certain inorganic materials, which are capable of converting low-energy infrared radiation into a higher-energy emission at visible wavelengths. This anti-Stokes shift enables luminescence detection without autofluorescence, which makes the upconverting materials a highly suitable reporter technology for optical biosensing applications. Furthermore, they exhibit long luminescence lifetime with narrow bandwidths also at the optical window of biomaterials enabling luminescence measurements in challenging sample matrices, such as whole blood. The aim of this thesis was to study the unique properties and the applicability of nano-sized upconverting phosphors (UCNPs) as reporters in biosensing applications. To render the inorganic nanophosphors water-dispersible and biocompatible, they were subjected to a series of surface modifications starting with silica-encapsulation and ending with a bioconjugation step with an analyte-recognizing biomolecule. The paramagnetism of the lanthanide dopants in the nanophosphors was exploited to develop a highly selective separation method for the UCNP-bioconjugates based on the magnetic selectivity of the high gradient magnetic separation (HGMS) system. The applicability of the nano-sized UCNPs as reporters in challenging sample matrices was demonstrated in two homogeneous sensing applications based on upconversion resonance energy transfer (UC-RET). A chemosensor for intracellular pH was developed exploiting UC-RET between the UCNP and a fluorogenic pH-sensitive dye with strongly increasing fluorescence intensity in decreasing pH. The pH-independent emission of the UCNPs at 550 nm was used for referencing. The applicability of the pH-nanosensor for intracellular pH measurement was tested in HeLa cells, and the acidic pH of endosomes could be detected with a confocal fluorescence microscope. Furthermore, a competitive UC-RET-based assay for red blood cell folic acid was developed for the measurement of folate directly from a whole blood sample. The optically transparent window of biomaterials was used in both the excitation and the measurement of the UC-RET sensitized emission of a near-infrared acceptor dye to minimize sample absorption, and the anti-Stokes detection completely eliminated the Stokes-shifted autofluorescence. The upconversion photoluminescence efficiency is known to be dependent on crystallite size, because the increasing surface-to-volume ratio of nano-sized UCNPs renders them more susceptible to quenching effects of the environment than their bulk counterpart. Water is known to efficiently quench the luminescence of lanthanide dopants. In this thesis, the quenching mechanism of water was studied using luminescence decay measurements. Water was found to quench the luminescence of UCNPs by increasing the non-radiative relaxation of the excited state of Yb3+ sensitizer ion, which had a very strong quenching effect on upconversion luminescence intensity.

Discrimination of Brazilian red wines according to the viticultural region, varietal, and winery origin

This work evaluated the physicochemical composition of 171 red Brazilian wines from the 2006 vintage, which were represented by 21 varietals. These wines were produced by 58 Brazilian wineries in different regions of the country, with latitudes varying from 9º to 31º South. Physicochemical wine analysis was performed in the same year and discrimination in the viticultural regions, varietal wines, and wineries was performed by means of the principal component analysis (PCA). The main results show that wines from São Joaquim had higher values of A420, A520, A620, color intensity, total phenolic compounds, anthocyanins, and dry extracts, while those from Toledo had lower values of these variables; those from Vale do São Francisco had higher values of potassium, pH, density, and volatile acidity; from Serra do Nordeste A, they had higher titratable acidity; and from Planalto Superior B, higher hue. Regarding the varietal wines, PCA mainly discriminated the wines produced from the varieties Ancellotta, Teroldego, Egiodola, Refosco, Marselan, Cabernet Sauvignon, Pinotage, Pinot Noir, Malbec, Arinarnoa, Barbera, and Alfrocheiro. In relation to wineries, twenty two of them were discriminated by their higher values of some variables, i.e., three were characterized by color intensity; three by hue; eight by alcohol content; six by potassium, dry extract, density, and pH; and two by titratablel acidity.

Evaluation of phenolic compounds content and in vitro antioxidant activity of red wines produced from Vitis labrusca grapes

Wine production in the northern Curitiba, Paraná, Brazil, specifically the communes of Colombo and Almirante Tamandaré, is based mainly on the utilization of Vitis labrusca grapes var. Bordô (Ives). Total sugar content, pH, and total acidity were analyzed in red wine samples from 2007 and 2008 vintages following official methods of analysis. Moreover, total phenolic, flavonoid, and tannin contents were analyzed by colorimetric methodologies and the antioxidant activity was determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical methodology. Phenolic compounds were identified by high performance liquid chromatography. The total phenolic content of wine samples presented concentrations varying between 1582.35 and 2896.08 mg gallic acid.L-1 since the major part corresponds to flavonoid content. In these compounds' concentration range, a direct relationship between phenolic compounds content and levels of antioxidant activity was not observed. Among the identified phenolic compounds, chlorogenic, caffeic, and syringic acids were found to be the major components. Using three principal components, it was possible to explain 81.36% of total variance of the studied samples. Principal Components Analysis does not differentiate between vintages.

Conversion of hydroxycinnamic acids into volatile phenols in a synthetic medium and in red wine by Dekkera bruxellensis

The conversion of p-coumaric acid, ferulic acid, and caffeic acid into 4-ethylphenol, 4-ethylguaiacol and 4-ethylcatechol was studied in Dekkera bruxellensis ISA 1791 under defined conditions in a synthetic medium and in a red wine. Liquid chromatography (HPLC-DAD) was used to quantify the phenolic acids, and gas chromatography (GC) coupled to a FID detector was used to quantify volatile phenols using a novel analytical methodology that does not require sample derivatization. Identification was achieved by gas chromatography-mass detection (GC-MS). The results show that phenolic acids concentration decreases while volatile phenols concentration increases. The proportion of caffeic acid taken up by Dekkera bruxellensis is lower than that for p-coumaric or ferulic acid; therefore less 4-ethylcatechol is formed. More important, 4-ethylcathecol synthesis by Dekkera bruxellensis in wine has never been demonstrated so far. These results contribute decisively to a better understanding of the origin of the volatile phenols in wines. The accumulation of these compounds in wine is nowadays regarded as one of the key factors of quality control.

Citotoxicity of food dyes sunset yellow (E-110), bordeaux red (E-123), and tatrazine yellow (E-102) on Allium cepa L. root meristematic cells

The objective of this study was to evaluate the cytotoxic effect of the food dyes sunset yellow, bordeaux red, and tartrazine yellow on the cellular cycle of Allium cepa L. Each dye was evaluated at the doses of 0.4 and 4.0 mL, at the exposure times of 24 and 48 hours in root tip cells of Allium cepa L. Slides were prepared and cells were analyzed during the whole cell cycle for cellular aberrations totaling 5,000 total cells for each dose evaluated. The mitotic index was calculated, and statistical analysis was performed using the Chi-squared test (p < 0.05). The results showed that the three dyes used under the evaluated doses and exposure times were cytotoxic to the cells of the system-test used. Further cytotoxicity studies should be conducted for additional results and a proper evaluation of the effect of these three dyes on a cellular level.

Mineral characterization of native fruits from the southern region of Brazil

Although the greatest variety of Brazilian flora is in the Amazon region, the Southern region of Brazil also has an estimated number of at least 5,000 species of vascular native plants. These species have been neglected as potential food sources, remaining unknown and under-utilized and limiting the potential variety in the diet of Brazilians and other peoples. Therefore the aim of this study was to characterize the mineral composition and content present in seven native fruit species of Southern Brazil using inductively coupled plasma optical emission spectrometry (ICP-OES). The essential element concentrations in the fruit samples were higher or similar to the values reported for traditional fruits. The araticum-do-mato fruit samples had high concentrations of the elements Ca, K, and Cu, and trace elements such as Pb and Sr. Mandacaru-de-três-quinas had predominance of Ba, Bi, and Ga, and the essential elements Mg and Mn. Uvaia and guabiroba had the highest levels of Al and Cr, but uvaia had high levels of Fe and Zn. The pindo palm had high amounts of Cd and Ni, and the yellow guava had high concentrations of Na, while red guava had high levels of Co.

Chloride concentration in red wines: influence of terroir and grape type

Red wines from different countries have been assessed in order to determine the influence of terroir and grape variety in their concentration of chloride. Chloride analysis was carried out by Laboratório de Bebidas de Origem Vegetal do Espírito Santo (Labeves), using the potentiometric method, in which the dosing is directly applied to the sample with an Ag/AgCl electrode. Data were collected to establish the level of chloride, as presented in the analysis reports issued by Labeves, and to serve as a wine categorization database, according to grape variety and country of origin. Australia and Argentina presented the highest levels of chloride and the wines made from the Syrah variety presented the highest concentration of such ion. We have, therefore, found that terroir and grape variety do have an influence over the concentration of chloride in red wines.

Chemical markers and antifungal activity of red propolis from Sergipe, Brazil

The aim of this study was to analyze the physicochemical properties and antifungal activities of the red propolis samples from Sergipe, Brazil, and also evaluate their variability throughout the year. The characterization of the hydroalcoholic extract (HPE) of the red propolis samples was performed monthly from October 2009 to September 2010. The concentrations of the bioactive compounds varied during the year, but their chromatographic profiles were similar. Four compounds were identified by comparison with authentic standards. Formononetin was one of the predominant compounds in all propolis extracts. In our study, it was observed that all the propolis samples inhibited the growth of Candida species. Multivariate analysis confirmed the variations in chemical composition and color of the HPEs throughout the year. The biological activities of the HPEs were statistically significant (p<0.05), and all samples exhibited antifungal properties.

Physicochemical and antioxidative properties of black, brown and red rice varieties of northern Thailand

Rice, the seed of Oryza species, is the major cereal crop in most of the developing countries. Nearly 95% of global rice production is done in Asian countries, and about half of the world’s population consumes it. Some speciality rices are not commonly consumed. Colored rice is one of such variety. In these varieties, high amounts of anthocyanin pigment are deposited in the rice coat to form its black (also known as purple), brown and red colors. Minimum studies are there to explain the properties of these rice varieties of Thailand. Thus, the current study was aimed to assess the physicochemical and antioxidative properties of three rice varieties (Chiang Mai Black rice, Mali Red rice and Suphanburi-1 Brown rice) of different cultivars of northern Thailand. Rice bran extracts of these three cultivars were prepared with different solvents (polar and non-polar) for the evaluation of total phytochemical content and anti-oxidant free-radical-scavenging properties. Chiang Mai Black rice contained higher concentration of phenolic acid, flavonoids, and anthocyanins (Cyanidin 3-glucoside, peonidin 3-glucoside, cyanidin chloride). Chiang Mai Black rice is richer in free-radical-scavenging compounds and activities than the other tested varieties. Polar extractions of rice bran are high in anti-oxidative compounds and activities than non-polar extractions.

Fresh meat quality of pigs fed diets with different fatty acid profiles and supplemented with red wine solids

Abstract Three groups of pigs were fed three different diets, namely a diet rich in saturated fatty acids (palm oil-based, PO), a polyunsaturated fatty acid (PUFA)-rich diet (corn oil-based, CO), and a PUFA-rich diet (corn oil-based) supplemented with red wine solids (RWS), which was added to the diet (CO+RWS) in order to assess the protective effect on the oxidative status of the pork meat. The addition of corn oil favourably modulates the FAs profile of the backfat, and to a lesser extent of the intramuscular fat of semimembranosus muscle, without causing adverse effects on the meat quality or on its oxidative stability. Moreover, these parameters were not affected by the addition of the RWS in the CO+RWS diet.

Optimization of the HS-SPME-GC/MS technique for determining volatile compounds in red wines made from Isabel grapes (Vitis labrusca)

Abstract Brazilian wine production is characterized by Vitis labrusca grape varieties, especially the economically important Isabel cultivar, with over 80% of its production destined for table wine production. The objective of this study was to optimize and validate the conditions for extracting volatile compounds from wine with the solid-phase microextraction technique, using the response surface method. Based on the response surface analysis, it can be concluded that the central point values maximize the process of extracting volatile compounds from wine, i.e., an equilibrium time of 15 minutes, an extraction time of 35 minutes, and an extraction temperature of 30 °C. Esters were the most numerous compounds found under these extraction conditions, indicating that wines made from Isabel cultivar grapes are characterized by compounds that confer a fruity aroma; this finding corroborates the scientific literature.

Quality indicators and shelf life of red octopus (Octopus maya) in chilling storage

Abstract There are no precedents concerning the quality of Octopus maya during chilled storage. This study evaluated the shelf life of the red octopus in chilling storage (4oC) and the correlation of the sensory quality index with microbiological counting and the biochemical indicators (hypoxanthine, histamine and volatile amines). A total of 112 whole raw octopi (average weight of 896 g) were randomly selected from seven batches and exposed to 4°C for 18, 24, 48, 72, 84, 96, and 100 h. The histamine concentration (91.7%), followed by the counts of psychrotrophic bacteria (5.5%) and hypoxanthine (2.2%), were the predictors from the redundancy analysis that better explained the changes taking place during the chilling hours. After 72 h of chilling, the microbial count was determined to be log 4.7 CFU/g, and the octopus samples were classified as B quality (minor sensory quality defects) based on the sensory quality scale. Although the samples were not classified as unacceptable at 100 h of refrigeration by the sensory index, the level of histamine reached the defect action level (5 mg/100 g) as ruled by the International Food Safety Authorities. The shelf life of the red octopus in chilling storage was predicted to be 119 h.

Little Red Riding Hood

Kuvitusta satuun Punahilkka.