Oxidant/antioxidant properties of Croatian native propolis

Native propolis was defined as propolis powder collected from the continental part of Croatia and prepared according to a patented process that preserves all the propolis natural nutritional and organoleptic qualities. Nine phenolic compounds (out of thirteen tested) in propolis sample were detected by high performance liquid chromatography (HPLC) analysis. Among them chrysin was the most abundant (2478.5 microg/g propolis). Contrary to moderate antioxidant activity of propolis examined in vitro (ferric reduction antioxidant power; FRAP-assay), propolis as a food supplement modulated antioxidant enzymes (AOE) and significantly decreased lipid peroxidation processes (LPO) in plasma, liver, lungs, and brain of mice. The effect was dose- and tissue-dependent. The lower dose (100 mg/kg bw) protected plasma from oxidation, whereas the higher dose (300 mg/kg bw) was pro-oxidative. Hyperoxia (long-term normobaric 100% oxygen) increased LPO in all three organs tested. The highest vulnerability to oxidative stress was observed in lungs where hyperoxia was not associated with augmentation of AOE. Propolis protected lungs from hyperoxia by increased catalase (CAT) activity. This is of special importance for lungs since lungs of adult animals are highly vulnerable to oxidative stress because of their inability to augment AOE activity. Because of its strong antioxidant and scavenging abilities, native propolis might be used as a strong plant-based antioxidant effective not only in physiological conditions but also in cases that require prolonged high concentration of oxygen.     

Development of a method for the genetic identification of mussel species belonging to Mytilus, Perna, Aulacomya, and other genera

Legislation regarding the labeling of processed products is an important issue in the protection of consumer rights. This labeling is especially important in products that cannot be identified on the basis of their morphological characters, because these are removed from the animal in the transformation process. The goal of this study was the identification of mussel species using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Forensically Informative Nucleotide Sequencing (FINS) methodologies. The molecular marker selected was 18S rDNA (nuclear small-subunit rDNA gene), which allows identification at the genus level and at the species level in some cases. The genera included in this study were Mytilus, Perna, Aulacomya, Semimytilus, Brachidontes, Choromytilus, and Perumytilus. Different markers were used for genetic identification at the species level. To identify the species included in the genus Perna and Choromytilus, a fragment of ITS 1 (Internal Transcribed Spacer 1) was amplified by multiplex PCR and digested with restrictases. The species of Mytilus were identified by length polymorphism and RFLP of the polyphenolic adhesive protein gene. This methodology was validated with products manufactured in the authors' pilot plant and applied to commercial samples. Therefore, this sequential method can be completely or partially used to determine the mussel genus or species present in any food product.     

Antibacterial activity of coffee extracts and selected coffee chemical compounds against enterobacteria

The in vitro antimicrobial activity of commercial coffee extracts and chemical compounds was investigated on nine strains of enterobacteria. The antimicrobial activity investigated by the disc diffusion method was observed in both the extracts and tested chemical compounds. Even though pH, color, and the contents of trigonelline, caffeine, and chlorogenic acids differed significantly among the coffee extracts, no significant differences were observed in their antimicrobial activity. Caffeic acid and trigonelline showed similar inhibitory effect against the growth of the microorganisms. Caffeine, chlorogenic acid, and protocatechuic acid showed particularly strong effect against Serratia marcescens and Enterobacter cloacae. The IC(50) and IC(90) for the compounds determined by the microtiter plate method indicated that trigonelline, caffeine, and protocatechuic acids are potential natural antimicrobial agents against Salmonella enterica. The concentrations of caffeine found in coffee extracts are enough to warrant 50% of the antimicrobial effect against S. enterica, which is relevant to human safety.     

Phenylphenalenone type compounds from the leaf fibers of abaca (Musa textilis)

A series of phenylphenalenone type compounds, known to play a role as phytoalexins in plants of the Musaceae family, have been identified for the first time in the leaf fibers of abaca (Musa textilis). Among the phenylphenalenone type compounds identified, the structure of a novel compound, (1R)-2,3-dihydro-4,9-dihydroxy-8-methoxy-1-phenylphenalene, has also been described in abaca fibers. Its structure was elucidated by analysis of one- and two-dimensional NMR (correlation spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond correlation) spectroscopic data.     

Production and composition of cider spirits distilled in "alquitara"

The capacity of alquitara (a traditional distillation system) to produce cider brandies is evaluated. To do so, the chemical composition of 12 fractions obtained during the distillation process and the cider brandies obtained from five ciders were analyzed (alcohol strength, methanol, volatile substances, furfural, and metals), taking into account European and Spanish legislation. During the course of distillation, an important increase in methanol, furfural, 2-phenylethanol, and metals in the last fractions was observed, while fusel oils were more abundant in the first fractions collected. Only acetaldehyde behaved differently, showing a minimum concentration in the middle fractions that might be explained by its formation on the surface of alquitara. On the other hand, the final distillates obtained by means of this method complied with the considered regulations. Worth highlighting in this regard are the low levels of a potential toxin such as methanol, as well as the detection of a constant ratio for methanol, ethanol, and fusel oil for the pairs of cider/spirits analyzed, which could be interpreted as an indication of good uniformity in the distillation system and method, thus guaranteeing product quality.     

Molecular cloning and characterization of sea bass (Dicentrarchus labrax L.) CD8alpha

In this work, the gene and cDNA of the sea bass CD8alpha have been isolated and characterized. The coding sequence has an ORF of 666 bp. It retains the Ig motif that interacts with MHC and the two cysteines responsible for an intra-chain disulfide bridge. The hinge region contains the two essential cysteines involved in dimerization. The transmembrane region is well conserved in all analysed sequences. Similar to other teleosts, the cytoplasmic region lacks the consensus p56(lck) motif common in higher vertebrates. Analysis of the expression pattern using RT-PCR shows the highest expression in the thymus. Like in the human gene, the sea bass CD8alpha genomic structure is organized into six exons, which roughly correspond to separate functional domains of the protein. Southern blotting shows that CD8alpha exists as a single copy gene. Together, these results support the concept that the basic structure of CD8alpha has been maintained through evolution.     

Control of the taeniosis/cysticercosis complex: future developments

Cysticercosis is due to the establishment of the larval stage of the zoonotic cestode parasite Taenia solium. The infection causes substantial human morbidity and mortality, particularly in several Latin American countries and parts of Africa and Asia, as well as economic losses in pig husban dry due to condemnation of infected pork meat. The life cycle of T. solium includes human beings as definitive hosts and pigs as intermediate hosts. Cysticercosis is acquired by the ingestion of eggs released by human tapeworm carriers, who become infected after ingesting pork meat contaminated with cysticerci. Taenia solium transmission has been associated with poverty, lack of sanitary services and practices of rearing backyard pigs with free access to the areas that villagers use as toilets, as well as cultural behaviour. Nonetheless, due to the recent increase of migration and tourism, industrial countries are also reporting cases of human cysticercosis. There are many epidemiological studies that have been conducted mainly in Latin American countries that have evaluated intervention measures for control of cysticercosis including the development and testing of vaccines. Furthermore, the involvement of international agencies and institutions, such as the World Health Organization, the Food and Agriculture Organization and the International Livestock Research Institute, as well as the commitment of policymakers, scientists and field workers, are key means for the sustainable control and, hopefully, eradication of T. solium infections.     

Molecular characterization of Haemophilus parasuis ferric hydroxamate uptake (fhu) genes and constitutive expression of the FhuA receptor

Bacteria have evolved a set of highly specialized proteins to capture iron in iron-depleted environments. The acquisition and uptake of iron present in the extracellular milieu of eukaryotic organisms is indispensable for the growth and survival of microbial pathogens in the course of infection. Haemophilus parasuis is the causative agent of Gl?sser disease, which is responsible for considerable financial losses in pig-rearing worldwide. To gain insight into the mechanisms involved in siderophore-mediated iron uptake in H. parasuis, genes in the H. parasuis ferric hydroxamate uptake (Fhu) region were amplified in the work being reported here. As has been described in A. pleuropneumoniae, an Fhu genomic region was also present in H. parasuis, being composed of four potential consecutive open reading frames (ORF) designated as fhuC, fhuD, fhuB, and fhuA, respectively. By immunoblotting, using a cross-reactive polyclonal antibody raised against Actinobacillus pleuropneumoniae FhuA protein, it was demonstrated that this protein was constitutively expressed in H. parasuis and its level of expression was not modified under conditions of restricted iron availability. This is the first report describing the presence of the fhu genes in H. parasuis. Our results indicate that FhuA protein expression is not affected under iron-restricted conditions, however, it is one of the targets of the humoral immune response.     

Carotenoid profile modification during refrigerated storage in untreated and pasteurized orange juice and orange juice treated with high-intensity pulsed electric fields

A comparative study was made of the evolution and modification of various carotenoids and vitamin A in untreated orange juice, pasteurized orange juice (90 degrees C, 20 s), and orange juice processed with high-intensity pulsed electric fields (HIPEF) (30 kV/cm, 100 micros), during 7 weeks of storage at 2 and 10 degrees C. The concentration of total carotenoids in the untreated juice decreased by 12.6% when the juice was pasteurized, whereas the decrease was only 6.7% when the juice was treated with HIPEF. Vitamin A was greatest in the untreated orange juice, followed by orange juice treated with HIPEF (decrease of 7.52%) and, last, pasteurized orange juice (decrease of 15.62%). The decrease in the concentrations of total carotenoids and vitamin A during storage in refrigeration was greater in the untreated orange juice and the pasteurized juice than in the juice treated with HIPEF. During storage at 10 degrees C, auroxanthin formed in the untreated juice and in the juice treated with HIPEF. This carotenoid is a degradation product of violaxanthin. The concentration of antheraxanthin decreased during storage, and it was converted into mutatoxanthin, except in the untreated and pasteurized orange juices stored at 2 degrees C.