Long-term habituation (LTH) in the crab Chasmagnathus: a model for behavioral and mechanistic studies of memory

A decade of studies on long-term habituation (LTH) in the crab Chasmagnathus is reviewed. Upon sudden presentation of a passing object overhead, the crab reacts with an escape response that habituates promptly and for at least five days. LTH proved to be an instance of associative memory and showed context, stimulus frequency and circadian phase specificity. A strong training protocol (STP) (³15 trials, intertrial interval (ITI) of 171 s) invariably yielded LTH, while a weak training protocol (WTP) (£10 trials, ITI = 171 s) invariably failed. STP was used with a presumably amnestic agent and WTP with a presumably hypermnestic agent. Remarkably, systemic administration of low doses was effective, which is likely to be due to the lack of an endothelial blood-brain barrier. LTH was blocked by inhibitors of protein and RNA synthesis, enhanced by protein kinase A (PKA) activators and reduced by PKA inhibitors, facilitated by angiotensin II and IV and disrupted by saralasin. The presence of angiotensins and related compounds in the crab brain was demonstrated. Diverse results suggest that LTH includes two components: an initial memory produced by spaced training and mainly expressed at an initial phase of testing, and a retraining memory produced by massed training and expressed at a later phase of testing (retraining). The initial memory would be associative, context specific and sensitive to cycloheximide, while the retraining memory would be nonassociative, context independent and insensitive to cycloheximide

Techniques of radioautography for medical and biological research

Standard techniques for radioautography used in biological and medical research can be classified into three categories, i.e., macroscopic radioautography, light microscopic radioautography and electron microscopic radioautography. The routine techniques used in these three procedures are described. With regard to macroscopic radioautography, whole body radioautography is a standard technique which employs freezing and cryosectioning and can demonstrate organ distributions of both soluble and insoluble compounds. In contrast, in light and electron microscopic radioautography, soluble and insoluble techniques are separated. In order to demonstrate insoluble labeled compounds, conventional chemical fixations such as formalin for light microscopy or buffered glutaraldehyde and osmium tetroxide for both light and electron microscopy followed by dehydration, embedding and wet-mounting applications of radioautographic emulsions can be used. For the demonstration of soluble labeled compounds, however, cryotechniques such as cryofixation, cryosectioning, freeze-drying, freeze-substitution followed by dry-sectioning and dry-mounting radioautography should be employed both for light and electron microscopy. The outlines of these techniques, which should be utilized in various fields of biological and medical research, are described in detail

CYP1A1 and GSTP1 polymorphisms in an oral cancer case-control study

CYP1A1 and GSTP1 polymorphisms have been associated with a higher risk to develop several cancers, including oral squamous cell carcinoma (OSCC), which is closely related to tobacco and alcohol consumption. Both genes code for enzymes that have an important role in activating or detoxifying carcinogenic elements found in tobacco and other compounds, and polymorphic variants of these genes may result in alterations of the enzymatic activity. The CYP1A1 gene codes for the enzyme aryl hydrocarbon hydroxylase, which is responsible for the metabolism of polycyclic aromatic hydrocarbons. The investigated polymorphism, Ile/Val, seems to increase the activity of the enzyme in homozygous individuals, leading to an accumulation of carcinogens. The Ile/Val polymorphism occurs because of an A->G transition at exon 7, resulting in the CYP1A1*2B allele. The GSTP1*B variant shows an A->G transition at exon 5, changing the amino acid Ile to Val, with a reduced catalytic activity of the enzyme. Due to this reduction, the carriers of mutant alleles lost the capability to metabolize carcinogens, which could be responsible for a higher susceptibility to cancer. We conducted a case-control study in a group of 72 cases with newly diagnosed OSCC and 60 healthy controls matched for age, gender, smoking habits, and ethnicity. We used PCR methods to identify the allelic variants CYP1A1*2B and GSTP1*B. The data obtained showed no statistically significant association of allelic or genotypic variants of CYP1A1*2B (OR = 1.06; 95% CI = 0.49-2.29) and GSTP1*B (OR = 1.40; 95% CI = 0.70-2.79) with OSCC.

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.

β-carotene biotransformation to obtain aroma compounds

Carotenoids are important constituents of food due to their color and because their degradation products generate important volatile compounds in foods. Aroma compounds derived from carotenoids are widely distributed in nature, and they are precursors of many important aromas in foods such as fruits and in flowers as well. They present high aromatic potential and are therefore of great interest to the industries of aromas and fragrances. In this study, more than 300 previously isolated microorganisms with potential for biotransformation of β-carotene present in the culture medium were selected using the plate method; about 80 strains presented capacity to produce aroma compounds and 7 strains were selected by an untrained panel of tasters to generate aroma compounds. The β-ionone was the main compound produced by CS1 (34.0 mg.L-1) and CF9 (42.4 mg.L-1) microorganisms at 72 and 24 hours of fermentation, cultured with and without pre-inoculation, respectively. The β-damascone and pseudoionone were found in low concentrations, 1,1,6-trimethyl-1,2,3,4-tetrahydronaphthalen (TTN) was tentatively identified and other compounds such as apocarotenoids, apparently obtained from the cleavage of the central part of the carotenoid, were detected.

Microbial profile of a kefir sample preparations: grains in natura and lyophilized and fermented suspension

Probiotics are supplementary foods developed by microbial strains that improve animal health beyond basic nutrition. Probiotics are consumed orally, regardless of being considered as normal inhabitants of the intestines, able to survive in enzimatic and biliary secretions. Kefir is a probiotic originated from the old continent, fermented by several bacteria and yeasts, encapsulated in a polyssacharide matrix, and resembles jelly grains. Kefir is also presented as its sourish product both in sugary or milky suspensions containing vitamins, aminoacids, peptides, carbohydrates, ethanol, and volatile compounds. Kefir is known to have a diverse microbial content depending on the country and fermentative substrates, which cause distinct probiotic effects. In this sense, the purpose of this work was to isolate, identify, and quantify the microbial content of a native sugary kefir sample (fermented suspension and lyophilized natural grains). Serial dilutions were plated on Rogosa agar (AR) and De Man, Rogosa and Sharpe (MRS), for Lactobacillus; Brain Heart Infusion (BHI), for total bacteria; Sabouraud-Dextrose-Agar (SDA), for yeasts and filamentous fungi; Thioglycolate Agar (TA), for Streptococcus, Acetobacteria and Leuconostoc; and Coconut Water Agar (CWA), and CWA supplemented with yeast extract (CWAY), for various genera. Genera and species for all strains were identified through biochemical reactions and specific API systems. The microbial profile of kefir was different from other sources of grains despite the presence of similar microorganisms and others which have not been reported yet. The data obtained with the CWA and CWAE media suggest that both substrates are alternative and salutary media for culture of kefir strains.

Chemical pattern of brazilian apples: a chemometric approach based on the Fuji and Gala varieties

The chemical composition of apple juices may be used to discriminate between the varieties for consumption and those for raw material. Fuji and Gala have a chemical pattern that can be used for this classification. Multivariate methods correlate independent continuous chemical descriptors with the categorical apple variety. Three main descriptors of apple juice were selected: malic acid, total reducing sugar and total phenolic compounds. A chemometric approach, employing PCA and SIMCA, was used to classify apple juice samples. PCA was performed with 24 juices from Fuji and Gala, and SIMCA, with 15 juices. The exploratory and predictive models recognized 88% and 64%, respectively, as belonging to a mixed domain. The apple juice from commercial fruits shows a pattern related to cv. Fuji and Gala with boundaries from 0.18 to 0.389 g.100 mL-1 (malic acid), from 8.65 to 15.18 g.100 mL-1 (total reducing sugar) and from 100 to 400 mg.L-1 (total phenolic compounds), but such boundaries were slightly shorter in the remaining set of commercial apple juices, specifically from 0.16 to 0.36 g.100 mL-1, from 9.25 to 15.5 g.100 mL-1 and from 180 to 606 mg.L-1 for acidity, reducing sugar and phenolic compounds, respectively, representing the acid, sweet and bitter tastes.

Changes in breaded chicken and oil degradation during discontinuous frying with cottonseed oil

Discontinuous frying of breaded chicken in cottonseed oil was evaluated. Three 400 g batches of foodstuff were fried daily in a 28 L fryer at 182 °C for 4.5 minutes for 7-8 days, and the experiment was repeated three times. The total polar compounds in the oil were determined by the conventional method. Changes in the oil were determined by the quick tests Testo 265, Viscofrit and Fri-check based on physical constants, and the results were compared with those of total polar compounds obtained by the conventional method. The free fatty acids, conjugated dienes, Lovibond color, oxidative stability, fatty acid composition, and polymeric compounds were also determined. During frying, the oil samples presented 6.0-39.2% total polar compounds, 0.0-12.9% polymerized triacylglycerols, 1.3-14.5% oxidized triacylglycerols, 2.8-11.0% diacylglycerols, and 1.6-2.6% fatty acids and unsaponifiable polar compounds. The breaded chicken samples lost moisture, absorbed oil up to approximately 6%, and there were small changes in the fatty acid composition and low formation of trans-isomers. The best method for monitoring and discarding the oil was that used for the determination of total polar compounds.

Nutritional composition of Annona crassiflora pulp and acceptability of bakery products prepared with its flour

Annona crassiflora offers an edible fruit native to the Savanna. This study aimed to develop a flour meal from Annona crassiflora pulp; analyze the chemical composition of the fresh pulp and its flour; develop and verify the acceptance of formulations with different concentrations of the flour of Annona crassiflora pulp. Fruit used were selected and processed. The pulp was dried in an oven at 60-65 ºC/48h. We analyzed the chemical composition, and two formulations of breads were prepared with 10 and 20% Annona crassiflora pulp. The results showed that the drying of Annona crassiflora pulp enriched its nutritional value. The Annona crassiflora pulp showed important chemical components, as insoluble fibers (pulp and flour), minerals (potassium, calcium, manganese and others) and antioxidant compounds. The formulations were well-accepted in a sensory point of view and proved to be a good alternative to the exploitation of the fruit.

Chemical constituents and technological functional properties of acerola (Malpighia emarginata DC.) waste flour

Acerola is a fruit that can be consumed in the form of juice and pulp. However, during its processing, a large amount of waste is generated (seed and bagasse). Adding value to these by-products is of great interest, since their use can enrich foods with nutrients and fiber. In this study, we performed phytochemical screening, determined the proximate and mineral composition, bioactive compounds and the technological functional properties of acerola seed flour and acerola bagasse flour. Seeds were dried in a ventilated oven at ± 45 °C and the bagasse was lyophilized. Samples were ground, stored in flasks protected from light. Phytochemical screening revealed metabolites of nutritional and pharmacological interest and no potentially toxic substances in the flours. Seed flour and bagasse flour showed high levels (g 100 g- 1 of dry matter - DM) of soluble fiber: 4.76 and 8.74; insoluble fiber: 75.76 and 28.58, and phenolic compounds: 4.73 and 10.82, respectively. The flours also showed high absorption of water, oil and emulsion stability, presenting potential for inclusion in meat products and bakery products.

Changes in the physicochemical and microbiological properties of frozen araça pulp during storage

Araça belongs to the Myrtaceae family and is popularly known as araçá-comum, araçá-azedo, or araçá-do-campo. Frozen fruit pulp is of great importance for the food industry, which can produce it at the time of harvest, store it, and use it according to the demand of the consumer market and/or as an ingredient in the formulation of products such as yogurt, candies, and ice creams among others. The aim of this study was to evaluate the physical, chemical, and microbiological stability of frozen araça pulp during 12 months of frozen storage. It was observed that the levels of moisture (90.55-88.75%), ash (0.34-0.26%) total soluble sugars (7.11-6.62%), sucrose (3.55-1.39%), soluble pectin (0.24-0.23%), total pectin (0.5-0.46%), pH (3.82-2.31%), organic acids (698.12-122.25 µg.g-1 citric acid), and phenolic compounds (6.22-0.00 mg GAE.100 g-1) decreased during storage, whereas the levels of protein (0.61-0.83%), lipids (0.14-0.38%), total carbohydrates (8.36-9.78%), calorific value (37.14-45.86 kcal.100 g-1), reducing sugars (3.51-5.21%), soluble solids (5.17-6.0%), total antioxidant capacity (6.89-35.13%), and color parameters (L*49.75-50.67; a*0.79-1.82 and b*22.5-25.19) increased over the one-year storage period. According to the chemical and microbiological parameters assessed, the product can be stored for 12 months without loss of quality with addition of citric acid as a preservative.

Prevention of enzymatic browning of yacon flour by the combined use of anti-browning agents and the study of its chemical composition

Yacon roots present functional properties because of the high levels of fructooligosaccharides (FOS), which are considered as prebiotic fibers. In addition, yacon roots are rich in phenolic compounds. During the processing of yacon, the freshly cut surface undergoes rapid enzymatic browning. Control of enzymatic browning during processing is very important to preserve the appearance of yacon flour. In this study, it was evaluated the combined effect of anti-browning agents (ascorbic acid, citric acid and L-cysteine) on the inhibition of enzymatic browning of yacon, using Response Surface Methodology. The yacon pre-treated with anti-browning agents in concentrations of 15.0 mM for ascorbic acid, 7.5 mM for citric acid and 10.0 mM for L-cysteine was used for the processing of flour. Yacon flour presented an attractive color and good sensory properties, without residual aroma. The contents of FOS and phenolic compounds obtained in yacon flour were 28.60 g.100 g- 1 and 1.35 g.100 g- 1. Yacon flour can be considered as a potential functional food, especially due to high levels of FOS, which allows for its use in formulation of various foods.

Chemical characterization of the flour of peel and seed from two papaya cultivars

Papaya is among the currently most important tropical fruits grown in Brazil and in the world. The fruit is mainly consumed fresh although it offers many industrial products. The processing of this fruit, as well as its fresh consumption, results in large amounts of waste, such as peels and seeds. Papaya consumption is one of the causes of significant loss of food value; therefore, new aspects on the use of its waste as by-products, or in the production of food additives, or even the incorporation of its flour in food have aroused great interest because these are products of high nutritional value, and their use may be economically viable. The objective of this study was to produce and characterize peel and seed flours from two papaya cultivars (Havai and Calimosa) for their chemical constituents focusing on possible use in foods. The proximate and mineral composition, titratable acidity, soluble solids, pH, contents of vitamin C, and phenolic compounds were determined. According to the results obtained, the papaya peel and seed flours had high contents of protein and fiber and therefore can be used as alternative sources of nutrients and can also be added in foods avoiding waste and adding value to the fruit.

Evaluation of biological activities of Physalis peruviana ethanol extracts and expression of Bcl-2 genes in HeLa cells

Physalis species are used in folk medicine for phytotherapeutic properties. The extracts of medicinal plants are known to possess cytotoxic and chemopreventative compounds. In this study we investigated antibacterial, antioxidant, DNA damage preventative properties of Physalis peruviana (golden berry) on leaf and shoot ethanol extracts and their effects on cytotoxicity of HeLa cells and expression of apoptotic pathway genes. Among the tested bacteria for antibacterial activity, maximum inhibition zone was determined in Lactococcus lactis. The phenolic content was found higher in leaf extracts than shoot extracts. The antioxidant activity showed the highest TEAC values of the leaf (2 mg/mL) and the shoot (0.5 mg/mL) extracts as 0.291±0.04 and 0.192±0.015, respectively. In DNA damage prevention assay both leaf and shoot extracts, especially 30 and 20 µg/mL concentrations, exhibited significant protection against DNA damage-induced by hydroxyl radical generated by Fenton reaction. Our results suggest that leaf and shoot extracts possess cytotoxic effect on HeLa cells when applied as 100 µg/mL concentration. Also mRNA expression analysis showed the alteration of antiapoptotic genes, so the results suggest that P. peruviana ethanol extracts induce apoptotic cell death and should be investigated for identification of active compounds and their mechanisms of action.

Comparison between aroma compounds in wines from four Vitis vinifera grape varieties grown in different shoot positions

The aroma characteristics of wines from four Vitis vinifera grape varieties (‘Cabernet sauvignon’, ‘Merlot’, ‘Chardonnay’, and ‘Italian Riesling’) grown in three shoot positions were evaluated by HS-SPME-GC/MS. In this study, the numerous significant differences found in most of the aromatic compounds influence of different shoot positions on the quality of wine. The results showed that the middle shoot position increased significantly the aroma concentration in the majority of wines investigated. The volatile components showing the greatest differences in the wines of different cultivars were aldehydes and terpenes. 8 and 11 compounds were found and quantified (OAVs>1) in the two red wines and white wines at concentrations higher than their corresponding odor thresholds, respectively; and therefore they significantly contributed to the wine aromas. According to their OAVs, fruity, floral, cheese and fatty aroma strongly influenced the characteristics of the four monovarietal wines, while the two white wines showed the green and fresh aroma characteristics. These results are related to the different microclimate of the canopies of the different shoot positions and varieties. They suggest that proper elevating the fruiting zones could improve the accumulation of aroma compounds in wines from the different varieties. On the other hand, grapevines trained to systems with uniform fruiting zones could improve the quality of wine.