Effect of the 5-HT4 receptor and serotonin transporter on visceral hypersensitivity in rats

Visceral hypersensitivity plays an important role in motor and sensory abnormalities associated with irritable bowel syndrome, but the underlying mechanisms are not fully understood. The present study was designed to evaluate the expression of the 5-HT4 receptor and the serotonin transporter (SERT) as well as their roles in chronic visceral hypersensitivity using a rat model. Neonatal male Sprague-Dawley rats received intracolonic injections of 0.5% acetic acid (0.3-0.5 mL at different times) between postnatal days 8 and 21 to establish an animal model of visceral hypersensitivity. On day 43, the threshold intensity for a visually identifiable contraction of the abdominal wall and body arching were recorded during rectal distention. Histological evaluation and the myeloperoxidase activity assay were performed to determine the severity of inflammation. The 5-HT4 receptor and SERT expression of the ascending colon were monitored using immunohistochemistry and Western blot analyses; the plasma 5-HT levels were measured using an ELISA method. As expected, transient colonic irritation at the neonatal stage led to visceral hypersensitivity, but no mucosal inflammation was later detected during adulthood. Using this model, we found reduced SERT expression (0.298 ± 0.038 vs 0.634 ± 0.200, P < 0.05) and increased 5-HT4 receptor expression (0.308 ± 0.017 vs 0.298 ± 0.021, P < 0.05). Treatment with fluoxetine (10 mg·kg-1·day-1, days 36-42), tegaserod (1 mg·kg-1·day-1, day 43), or the combination of both, reduced visceral hypersensitivity and plasma 5-HT levels. Fluoxetine treatment increased 5-HT4 receptor expression (0.322 ± 0.020 vs 0.308 ± 0.017, P < 0.01) but not SERT expression (0.219 ± 0.039 vs 0.298 ± 0.038, P = 0.654). These results indicate that both the 5-HT4 receptor and SERT play a role in the pathogenesis of visceral hypersensitivity, and its mechanism may be involved in the local 5-HT level.

An interleukin-33/ST2 signaling deficiency reduces overt pain-like behaviors in mice

Interleukin (IL)-33, the most recent member of the IL family of cytokines, signals through the ST2 receptor. IL-33/ST2 signaling mediates antigen challenge-induced mechanical hyperalgesia in the joints and cutaneous tissues of immunized mice. The present study asked whether IL-33/ST2 signaling is relevant to overt pain-like behaviors in mice. Acetic acid and phenyl-p-benzoquinone induced significant writhing responses in wild-type (WT) mice; this overt nociceptive behavior was reduced in ST2-deficient mice. In an antigen-challenge model, ST2-deficient immunized mice had reduced induced flinch and licking overt pain-like behaviors. In the formalin test, ST2-deficient mice also presented reduced flinch and licking responses, compared with WT mice. Naive WT and ST2-deficient mice presented similar responses in the rota-rod, hot plate, and electronic von Frey tests, indicating no impairment of motor function or alteration in basal nociceptive responses. The results demonstrate that IL-33/ST2 signaling is important in the development of overt pain-like behaviors.

A method for fast determination of epigallocatechin gallate (EGCG), epicatechin (EC), catechin (C) and caffeine (CAF) in green tea using HPLC

Tea has been considered a medicine and a healthy beverage since ancient times, but recently it has received a great deal of attention because of its antioxidant properties. Green tea polyphenols have demonstrated to be an effective chemopreventive agent. Recently, investigators have found that EGCG, one of the green tea catechins, could have anti-HIV effects when bound to CD4 receptor. Many factors can constitute important influences on the composition of tea, such as species, season, age of the leaf, climate, and horticultural practices (soil, water, minerals, fertilizers). This paper presents an HPLC analytical methodology development, using column RP-18 and mobile phase composed by water, acetonitrile, methanol, ethyl acetate, glacial acetic acid (89:6:1:3:1 v/v/v/v/v) for simultaneous determination and quantification of caffeine (CAF), catechin (C), epicatechin (EC) and epigallocatechin gallate (EGCG) in samples of Camellia sinensis (green tea) grown in Brazil and harvested in spring, in summer and in autumn, in comparison to Brazilian black tea, to samples of Japanese and Chinese green tea and to two standardized dry extracts of green tea. The method has been statistically evaluated and has proved to be adequate to qualitative and quantitative determination of the samples.

Study of bacteria Gluconobacter sp.: isolation, purification, phenotypic and molecular identification

The importance of the study of acetic bacteria, on species of the Gluconobacter genus is based on its industrial application, as these possess the capacity of bioconversion of sorbitol to sorbose, enabling the process of vitamin C production. The study involved samples collected in industries of soft drinks, flowers, fruits and honey, followed by purification, phenotypic identification, molecular identification with the use of primer defined from Nucleotide Sequence Database consultation. Strains preserved were identified as members of the Acetobacteraceae family, Gluconobacter genus. 110 strains had been isolated of substrate: Pyrostegia venusta (ker-gawler), honey, Vitis vinifera (grape), Pyrus communis (pear), Malus sp. (apple) and in two samples of soft drinks. Of this total 57 strains had been recovered in manitol medium (manitol, yeast extract, peptone), 12 in YMG medium (glucose, manitol, yeast extract, ethanol, acetic acid), 41 in enrichment medium (De Ley and Swings) and later in the GYC medium (glucose, yeast extract and calcium carbonate). 68 strains were identified as Gram negative bacilli rods. Of these, 31 were characterized biochemically as belonging to the Acetobacteriaceae family as they were catalase positive, oxidase negative and producers of acid from glucose. The characterization of these strains was complemented with the biochemistry tests: gelatin liquefaction, nitrate reduction, indole and H2S production, oxidation of ethanol to acetic acid and molecular tests for genus identification. Only eight strains were characterized as pertaining to the Gluconobacter genus. The strains are maintained in collection cultures at the Microbiology Laboratory of the Biology Department at the São Paulo State University (UNESP) in Assis, stored in malt extract at -196 ºC.

Changes in guava (Psidium guajava L. var. Paluma) nectar volatile compounds concentration due to thermal processing and storage

Guava nectars were formulated for approximately 10, 12, or 14 ºBrix, with 40% guava pulp. Sodium benzoate, 500 mg.kg-1 was used as preservative. The Brix value was adjusted with saturated sucrose syrup. The guava nectar was pasteurized (85 ºC/42 seconds) in tubular heat exchanger and then hot filled in 500 mL white glass bottles. The products were stored either at room temperature (25 ± 5 ºC) or refrigerated (5 ± 2 ºC) under fluorescent light exposure and analyzed on the day after processing (time zero) and also 40, 80, and 120 days of storage. Eight compounds were identified and quantified by Gas Chromatography (GC) -Mass Spectrometry (MS): hexanal, (E)-hex-2-enal, 1-hexenol, (Z)-hex-3-enol, (Z)-hex-3-enyl acetate, phenyl-3-propyl acetate, cinnamyl acetate, and acetic acid. There was no significant effect of thermal treatment on the volatile compound concentrations, except for a significant decrease (p = 0.0001) in hexanal and (Z)-hex-3-enyl acetate (p = 0.0029). As for the storage time, there was a much greater decrease in the esters contents, such as (Z)-hex-3-enyl and phenyl-3-propyl acetates. Cinnamyl acetate had the greatest decrease over storage time. Refrigeration was better than room temperature for guava nectar volatile compounds stability over storage time, mainly for esters compounds, which are important for the product aroma and flavor

Determination of structural and mechanical properties, diffractometry, and thermal analysis of chitosan and hydroxypropylmethylcellulose (HPMC) films plasticized with sorbitol

In this work, the structural, mechanical, diffractometric, and thermal parameters of chitosan-hydroxypropylmethylcellulose (HPMC) films plasticized with sorbitol were studied. Solutions of HPMC (2% w/v) in water and chitosan (2% w/v) in 2% acetic acid solution were prepared. The concentration of sorbitol used was 10% (w/w) to both polymers. This solutions were mixed at different proportions (100/0; 70/30; 50/50; 30/70, and 0/100) of chitosan and HPMC, respectively, and 20 mL was cast in Petri dishes for further analysis of dried films. The miscibility of polymers was assessed by X-ray diffraction, scanning electronic microscopy (SEM), differential scanning calorimetry (DSC), and thermal gravimetric analysis (TGA). The results obtained indicate that the films are not fully miscible at a dry state despite the weak hydrogen bonding between the polymer functional groups.

Retention of short chain fatty acids under drying and storage conditions

Cheese whey permeate was used as a substrate for the fermentation of Propionibacterium freudenreichi PS1 for the production of short chain fatty acids, components of the bio-aroma of Swiss cheese. The liquid bio-aroma was encapsulated by spray drying under different conditions of air inlet temperature and feed rate. A study was carried out on the stability of the bio-aroma during storage in laminated packages at 30 °C for 96 days using the product showing the greatest retention of acetic and propionic acids. The results showed that the best drying conditions were an air entrance temperature of 180 °C and a feed rate of 24 g/min resulting in particles with a smooth surface and few invaginations and micro-fissures. However, 72% of the acetic acid and 80% of the propionic acid were lost during storage showing that the wall material used was inadequate to guarantee product stability.

Effect of starter culture and inulin addition on microbial viability, texture, and chemical characteristics of whole or skim milk Kefir

The effect of inulin addition and starters (Kefir grains or commercial starter culture) on the microbial viability, texture, and chemical characteristics of Kefir beverages prepared with whole or skim milk was evaluated during refrigerated storage. The type of starter did not influence microbial viability during the storage of the beverages, but the chemical and textural changes (decreases in pH, lactose concentration, and inulin and increased acidity, firmness, and syneresis) were more pronounced in the formulations fermented with grains than those fermented with the starter culture. The addition of inulin did not influence acidity or viability of lactic acid bacteria, but in general, its effect on the survival of acetic acid bacteria, Lactococcus and yeasts, firmness, and syneresis depended on the type of milk and starter culture used. Generally, the yeast, acetic acid bacteria, and Leuconostoc counts increased or remained unchanged, while the total population of lactic acid bacteria and Lactococcus were either reduced by 1 to 2 logs or remained unchanged during storage.

Characterization and extraction of volatile compounds from pineapple (Ananas comosus L. Merril) processing residues

The aim of this study was to extract and identify volatile compounds from pineapple residues generated during concentrated juice processing. Distillates of pineapple residues were obtained using the following techniques: simple hydrodistillation and hydrodistillation by passing nitrogen gas. The volatile compounds present in the distillates were captured by the solid-phase microextraction technique. The volatile compounds were identified in a system of high resolution gas chromatography system coupled with mass spectrometry using a polyethylene glycol polar capillary column as stationary phase. The pineapple residues constituted mostly of esters (35%), followed by ketones (26%), alcohols (18%), aldehydes (9%), acids (3%) and other compounds (9%). Odor-active volatile compounds were mainly identified in the distillate obtained using hydrodistillation by passing nitrogen gas, namely decanal, ethyl octanoate, acetic acid, 1-hexanol, and ketones such as γ-hexalactone, γ-octalactone, δ-octalactone, γ-decalactone, and γ-dodecalactone. This suggests that the use of an inert gas and lower temperatures helped maintain higher amounts of flavor compounds. These data indicate that pineapple processing residue contained important volatile compounds which can be extracted and used as aroma enhancing products and have high potential for the production of value-added natural essences.

Stability of frozen marolo pulp during storage

Marolo, also known as araticum or head-to-black, is a globular berry, a species native to the Brazilian savannah. The aim of this study was to evaluate the physical, chemical, and microbiological stability of frozen marolo pulp during 12 months of frozen storage. It was observed that the levels of ash (0.28-0.22%), protein (0.77-0.71%), lipids (1.75-1.73%), carbohydrates (12.1-10.15%), calorie (67.23-59.01 kcal), sucrose (2.50-1.29%), citric acid (435.63-197.5 µg.g-1), tartaric acid (4.38-1.88 µg.g-1) , acetic acid (470.38-279.25 µg.g-1), ascorbic acid (3.00-0.00 µg.g-1), total pectin (0.67-0.39%), pH (3.88-3.83), and b* chromaticity coordinates (24.85-20.53) decreased reduced during storage, whereas the levels of moisture (85.10-87.19%), color parameters (L* 58.89-62.62 and a* 5.37-7.86), reducing sugars (4.53-5.62%), total soluble sugars (7.1-7.36%), soluble solids (7.0-8.4 ºBrix), total acidity (0.9-1.0%), malic acid (514.13-781.25 µg.g-1), soluble pectin (0.16-0.24%), and antioxidant (6.85-37.35% of DPPH discoloration) increased over the one-year of storage period. According to the physical, 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.

Antioxidant activities of kombucha prepared from three different substrates and changes in content of probiotics during storage

Kombucha is a health-promoting fermented beverage worldwide. The present study compared the free-radical scavenging abilities and total reducing power (TRP) of kombucha prepared from low-cost green tea (LGTK), black tea (BTK), and tea powder (TPK). LGTK had the highest scavenging abilities against 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide anion and hydroxyl radicals, while BTK showed the highest TRP. Changes in content of probiotics in LGTK were investigated during storage as well. The number of acetic acid bacteria decreased moderately up to 10 days of storage. The number of lactic acid bacteria (LAB) decreased significantly, and their survival rate was only 0.98% at the 8th day of storage.

Vinegar rice (Oryza sativa L.) produced by a submerged fermentation process from alcoholic fermented rice

Considering the limited availability of technology for the production of rice vinegar and also due to the potential consumer product market, this study aimed to use alcoholic fermented rice (rice wine (Oryza sativa L.)) for vinegar production. An alcoholic solution with 6.28% (w/v) ethanol was oxidized by a submerged fermentation process to produce vinegar. The process of acetic acid fermentation occurred at 30 ± 0.3°C in a FRINGS® Acetator (Germany) for the production of vinegar and was followed through 10 cycles. The vinegar had a total acidity of 6.85% (w/v), 0.17% alcohol (w/v), 1.26% (w/v) minerals and 1.78% (w/v) dry extract. The composition of organic acids present in rice vinegar was: cis-aconitic acid (6 mg/L), maleic acid (3 mg/L), trans-aconitic acid (3 mg/L), shikimic + succinic acid (4 mg/L), lactic acid (300 mg/L), formic acid (180 mg/L), oxalic acid (3 mg/L), fumaric acid (3 mg/L) and itaconic acid (1 mg/L).

Determination of some quality properties of marinated sea bream (Sparus Aurata L., 1758) during cold storage

In this study, it was aimed to determine the effect of sea bream (Sparus aurata) marinated some quality properties during cold storage. The fillets of fish were immersed into brine including 3.5% acetic acid 11% salt in the ratio of 1: 1.5 (fish: marinate brine) for marination process. After the process of ripening, samples were grouped into two and packed in plastic containers; one being plain (in sunflower oil) and the other being sauced (sauced prepared with sunflower oil). During storage, sensory, crude protein, lipid, dry matter and crude ash, TBA, TVB-N, TMA-N and peroxide analyses were done periodically. According to results of 200 days of storage, TVB-N values of sea bream marinates packaged as plain and sauced were 15.86/14.89 mg/100g, TBA 7.06/7.99 mg MA/kg, TMA-N 2.97/3.12, mg/100g, the value of peroxide was 7.23/7.45 meq/kg respectively. According to chemical and sensory analyses results obtained in the study; it was concluded that sea bream marinates packaged as plain and sauced can be stored in +4 °C for 200 days.

Recovery and purification of anthocyanins from purple-blue potato

The aim of this work was to study techniques to extract and purify of anthocyanins from purple-blue potato. This topic was determined as a master’s thesis and it was done in collaboration with the Food Chemistry and Food Development Department of University of Turku and Department of Chemical and Process Engineering at Lappeenranta University of Technology. At first, purple-blue potatoes were pretreated in four types of boiled, raw, freeze-dried and dried boiled potato for extraction. They were mixed with aqueous acidified ethanol (ethanol:water:acetic acid 40%:53%:7% v/v) for conventional extraction. Boiled potato was selected as a best pretreated potato. Different ethanol concentration and extraction time were examined and the mixture of 80% in 24 h resulted in maximum anthocyanin content (132.23 mg/L). As conventional extraction method of anthocyanins was non-selective, some of impurities such as free sugars might accelerate anthocyanin degradation. Therefore, to obtain anthocyanins in purified form, adsorption as a promising selective method was used to recovery and isolate anthocyanins. It was carried out with six adsorbents. Among those, Amberlite XAD-7HP, a nonionic acrylic ester adsorbent, was found to have the best performance. In an adsorption column, flow rate of 3 mL/min was selected as the loading flow rate among four tested flow rates. Eluent volume and flow rate were 3 BV of aqueous acidified ethanol (75%, v/v) and 1 mL/min for desorption. The quantification of the total anthocyanin contents was performed by pH-differential method using UV-vis spectrophotometer. The resulting anthocyanin solution after purification was almost free from free sugars which were the major cause for degradation of anthocyanins. The average anthocyanin concentration in the purified and concentrated sample was obtained 1752.89 mg/L.

Critical levels of organic acids on seed germination and seedling growth of wheat

The objective of this study was to determine the responses of the wheat cultivars CD 108 and CD 111 for tolerance to organic acids. The effects of five concentrations of the three main acids formed in the soil were studied: acetic acid (0, 4, 8, 12 and 16 mM), propionic acid (0, 4, 8, 12 and 16 mM) and butyric acid (0, 2, 4, 8 and 12 mM). Tests included germination, shoot length, root length and dry weight of shoot and root. The variable root length is the most responsive variable for all the acids tested and the critical level of toxicity of acetic, propionic and butyric acids, which reduced root length by at least 50% was 9.0, 8.5 and 4.0 mM respectively. It was concluded that the presence of acetic, propionic and butyric acids in the germination substratum of wheat seeds of the cultivars CD 111 and CD 108 reduced seedling development, mainly by reducing the length of the radicles.