The P2X7 purinergic receptor on bovine macrophages mediates mycobacterial death

P2X7 is an ATP gated purinoceptor that has been linked to various immune responses. P2X7 appears to be expressed ubiquitously in the immune system and thus may be important as an effector pathway or play significant roles in cell activation/death. 2',3'-(4-Benzoyl)benzoyl ATP is the most potent agonist of this receptor and ATP in its fully dissociated form (ATP(4-)) also activates the receptor. High concentrations of ATP can cause the P2X7 receptor to induce pore formation on the surface of the cell that allows molecules of considerable size to pass and can lead to cell death. The P2X7 receptor has also been linked to various immune activities when the concentration of ATP is lower, including the release of IL-1beta. The role P2X7 receptors have on immune cell activities is just beginning to be understood. We sought to determine the role of P2X7 on bovine macrophages in eliminating the causative agent of bovine-type tuberculosis, Mycobacterium bovis. Because high concentrations of ATP are linked to macrophage death, we determined if this method of cell destruction also leads to reduced bacterial viability. We find that P2X7 is present on bovine macrophages from different sources, including both peripheral blood-derived as well as alveolar macrophages. In addition, P2X7 mRNA is present in B and T lymphocytes. The treatment of M. bovis-infected macrophages with ATP results in reduced macrophage viability as well as reduced M. bovis viability.     

Evaluation of laryngeal mask as an alternative to endotracheal intubation in cats anesthetized under spontaneous or controlled ventilation

OBJECTIVE: To compare the cardiorespiratory effects and incidence of gastroesophageal reflux with the use of a laryngeal mask airway (LMA) or endotracheal tube (ET) in anesthetized cats during spontaneous (SV) or controlled ventilation (CV). STUDY DESIGN: Prospective randomized experimental trial. ANIMALS: Thirty-two adult crossbred cats, weighing 2.7 +/- 0.4 kg. METHODS: The cats were sedated with intramuscular (IM) methotrimeprazine (0.5 mg kg(-1)) and buprenorphine (0.005 mg kg(-1)), followed 30 minutes later by induction of anesthesia with intravenous (IV) thiopental (12.5-20 mg kg(-1)). An ET was used in 16 cats and an LMA in the remaining 16 animals. Anesthesia was maintained with 0.5 minimum alveolar concentration (0.6%) of halothane in oxygen using a Mapleson D breathing system. Cats in both groups were further divided into two equal groups (n = 8), undergoing either SV or CV. Neuromuscular blockade with pancuronium (0.06 mg kg(-1)) was used to facilitate CV. Heart and respiratory rates, direct arterial blood pressure, capnometry (PE'CO2) and arterial blood gases were measured. Gastric reflux and possible aspiration was investigated by intragastric administration of 5 mL of radiographic contrast immediately after induction of anesthesia. Cervical and thoracic radiographs were taken at the end of anesthesia. Data were analyzed using anova followed by Student-Newman-Keuls, Kruskal-Wallis or Friedman test where appropriate. RESULTS: Values for PaCO2 and PE'CO2 were higher in spontaneously breathing cats with the LMA when compared with other groups. Values of PaO2 and hemoglobin oxygen saturation did not differ between groups. Gastroesophageal reflux occurred in four of eight and two of eight cats undergoing CV with ET or LMA, respectively. There was no tracheal or pulmonary aspiration in any cases. CONCLUSIONS AND CLINICAL RELEVANCE: The use of an LMA may be used as an alternative to endotracheal intubation in anesthetized cats. Although aspiration was not observed, gastric reflux may occur in mechanically ventilated animals.     

Functional and Bioactive Properties of Peptides Derived from Marine Side Streams

In fish processing, a great amount of side streams, including skin, bones, heads and viscera, is wasted or downgraded as feed on a daily basis. These side streams are rich sources of bioactive nitrogenous compounds and protein, which can be converted into peptides through enzymatic hydrolysis as well as bacterial fermentation. Peptides are short or long chains of amino acids differing in structure and molecular weight. They can be considered as biologically active as they can contribute to physiological functions in organisms with applications in the food and pharmaceutical industries. In the food industry, such bioactive peptides can be used as preservatives or antioxidants to prevent food spoilage. Furthermore, peptides contain several functional qualities that can be exploited as tools in modifying food ingredient solubility, water-holding and fat-binding capacity and gel formation. In the pharmaceutical industry, peptides can be used as antioxidants, but also as antihypertensive, anticoagulant and immunomodulatory compounds, amongst other functions. On the basis of their properties, peptides can thus be used in the development of functional foods and nutraceuticals. This review focuses on the bioactive peptides derived from seafood side streams and discusses their technological properties, biological activities and applications.     

Development of Antioxidant Protein Extracts from Gilthead Sea Bream (Sparus aurata) Side Streams Assisted by Pressurized Liquid Extraction (PLE)

The pressurized liquid extraction (PLE) technique was used, for the first time, to obtain protein extracts with antioxidant activity from side streams (muscle, heads, viscera, skin, and tailfins) of gilthead sea bream (Sparus aurata) in order to give added value to these underutilized matrices. Extraction conditions previously optimized for sea bass (Dicentrarchus labrax) side streams were applied. Protein recovery percentages were 22% (muscle), 33% (heads), 78% (viscera), 24% (skin), and 26% (tailfins), which represented an increase of 1.2–4.5-fold compared to control samples (extraction by stirring). The SDS-PAGE profiles revealed that PLE-assisted extraction influenced protein molecular weight distribution of the obtained extracts. PLE conditions also allowed increasing the antioxidant capacity measured by both Trolox equivalent antioxidant capacity (TEAC; 1.3–2.4 fold) and oxygen radical absorbance capacity (ORAC; 1.9–6.4) assays for all fish extracts. Inductively coupled plasma mass spectrometry (ICP-MS) and high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-qTOF-MS) were used to investigate the presence of toxic metals and mycotoxins in sea bream side streams. The levels of As, Hg, Cd, and Pb were below those established by authorities for fish muscle for human consumption (except for Cd in viscera samples). Through a nontargeted screening approach, no mycotoxins or related metabolites were detected for all sea bream side streams. This study contributes to the research on the valorization of fish processing side streams using environmentally friendly technology.     

Salmon (Salmo salar) Side Streams as a Bioresource to Obtain Potential Antioxidant Peptides after Applying Pressurized Liquid Extraction (PLE)

The pressurized liquid extraction (PLE) technique was used to obtain protein extracts with antioxidant capacity from salmon muscle remains, heads, viscera, skin, and tailfins. A protein recovery percentage ≈28% was obtained for all samples except for viscera, which was ≈92%. These values represented an increase of 1.5–4.8-fold compared to stirring extraction (control). Different SDS-PAGE profiles in control and PLE extracts revealed that extraction conditions affected the protein molecular weight distribution of the obtained extracts. Both TEAC (Trolox equivalent antioxidant capacity) and ORAC (oxygen radical antioxidant capacity) assays showed an outstanding antioxidant activity for viscera PLE extract. Through liquid chromatography coupled with electrospray ionization triple time-of-flight (nanoESI qQTOF) mass spectrometry, 137 and 67 peptides were identified in control and PLE extracts from salmon viscera, respectively None of these peptides was found among the antioxidant peptides inputted in the BIOPEP-UMP database. However, bioinformatics analysis showed several antioxidant small peptides encrypted in amino acid sequences of viscera extracts, especially GPP (glycine-proline-proline) and GAA (glycine-alanine-alanine) for PLE extracts. Further research on the relationship between antioxidant activity and specific peptides from salmon viscera PLE extracts is required. In addition, the salmon side streams studied presented non-toxic levels of As, Hg, Cd, and Pb, as well as the absence of mycotoxins or related metabolites. Overall, these results confirm the feasible use of farmed salmon processing side streams as alternative sources of protein and bioactive compounds for human consumption.     

The high tolerance of different pomegranate cultivars to the excess of boron in irrigation water is due to their capacity to limit boron transport from the root to the leaves

Background : Presently, irrigation waters often have a high concentration of boron (B), and the fruit trees in the Mediterranean areas, in general, are exposed to a high risk of B toxicity. Aims : To test the hypothesis that pomegranate trees are very tolerant to the presence of B in the irrigation water, to elucidate the physiological mechanisms behind this tolerance, and to assess differences between different varieties. Methods : In this study, the physiological and nutritional behavior of three pomegranate cultivars (‘Mollar de Elche', ‘Valenciana', and ‘Wonderful'), treated with five B concentrations (0.25, 1.25, 2.5, 5.0, 10.0 mg L^?1), were studied. At day 30 and 60 after the start of the treatments, growth measurements, gas exchange parameters and organic solutes were recorded. Results : Almost no significant differences were observed between the B treatments for any of the cultivars. Increasing the B concentration in the nutrient solution increased B concentrations in the leaves, but they never exceed 40 mg kg^?1 of B. In the roots, however, B concentrations were very high, up to 400 mg kg^?1. Conclusion : Pomegranate trees are very tolerant to B excess due to their ability to accumulate B in great quantities in the root, without causing toxicity to this part of the plant, thus limiting its transport to the shoots.     

EFFECT OF THE APPLICATION OF MONOSILICIC ACID ON THE PRODUCTION AND QUALITY OF TRIPLOID WATERMELON

Experiments were conducted in two consecutive years in a multilayered plastic-covered greenhouse, of the “raspa and amagado” (covered skeleton structure) types in the fields of Almeria (south-eastern Spain), to evaluate the production and quality of a triploid watermelon crop (Citrullus lanatus Thunb) cv. ‘Queen of Hearts’ grafted onto the (Cucurbita maxima x Cucurbita moschata) squash hybrid cv. ‘Strongtosa’. A 2.5% solution of the monosilicic acid [Si(OH)₄] product was applied to this crop in the drip irrigation system at a dose of 260 mL·ha^?1 in both experiments. Results showed that the product did not influence productive parameters in terms of kg of product·m^?2, kg plant^?1, or number of fruits plant^?1. Nevertheless the monosilicic acid [Si(OH)₄] product appeared to have a positive effect on some quality aspects of the fruit with an approximate 10% increase in pulp consistency and rind width.