Chemical and microbiological quality of grass carp (Ctenopharyngodon idella) slaughtered by different methods

The effect of two slaughter methods (immersion in ice-water slurry and electrical stunning followed by ice slurry asphyxiation) on chemical and microbiological parameters of grass carp (Ctenopharyngodon idella) stored in ice for 20 days was evaluated. No differences in total volatile basic nitrogen (TVB-N), pH, carbohydrate or protein content of mucus were observed between the slaughter methods. Ice-slaughtered fish had lower bacteria counts at the beginning of storage, but higher counts than fish slaughtered by electricity at the end of storage (p < 0.05). However, no significant differences in the shelf life were observed between the slaughter methods evaluated (limit of acceptability – counts > 3 × 10⁶ CFU g⁻¹ – attained after 13–16 days). Results indicated that the chemical parameters evaluated have a limited applicability to assess the shelf life of grass carp stored in ice, since pH limit (6.8) was exceeded after 4 days, while TVB-N limit (30 mg%) was not attained after 20 days of storage.     

Characterization of nicotinamide mononucleotide adenylyltransferase from thermophilic archaea

The enzyme nicotinamide mononucleotide (NMN) adenylyltransferase (EC 2.7.7.1) catalyzes the synthesis of NAD+ and nicotinic acid adenine dinucleotide. It has been purified to homogeneity from cellular extracts of the thermophilic archaeon Sulfolobus solfataricus. Through a database search, a highly significant match was found between its N-terminal sequence and a hypothetical protein coded by the thermophilic archaeon Methanococcus jannaschii MJ0541 open reading frame (GenBank accession no. U67503). The MJ0541 gene was isolated, cloned into a T7-based vector, and expressed in Escherichia coli cells, yielding a high level of thermophilic NMN adenylyltransferase activity. The expressed protein was purified to homogeneity by a single-step chromatographic procedure. Both the subunit molecular mass and the N-terminal sequence of the pure recombinant protein were as expected from the deduced amino acid sequence of the MJ0541 open reading frame-encoded protein. Molecular and kinetic properties of the enzymes from both archaea are reported and compared with those already known for the mesophilic eukaryotic NMN adenylyltransferase.     

Trans Fat Supplementation Increases UV-Radiation-Induced Oxidative Damage on Skin of Mice

We evaluated the influence of fish oil (FO, rich in n-3 FA), soybean oil (SO, rich in n-6 FA) and hydrogenated vegetable fat (HVF, rich in trans FA) on the oxidative status and viability of skin cells of mice exposed to ultraviolet radiation (UVR). Mice were supplemented with FO, SO or HVF for three months and exposed to UVR (2.72 mJ/cm²) for 2 days. One day after the last UVR session, the FO group showed higher levels of n-3 fatty acids (FA), while the HVF showed higher incorporation of trans FA (TFA) in dorsal skin. UVR increased lipid peroxidation and protein carbonyl levels of the HVF and to a lesser extent of the control and SO groups. Although all irradiated groups showed increased skin thickness, this increase was slighter in FO mice. UVR exposure reduced skin cell viability of the control, SO and HVF groups, while FO prevented this. Catalase activity was reduced independently of the supplementation and SOD level was increased in C and FO groups after UVR exposure; FO prevented the UVR-induced increase in glutathione levels, which was observed in skin of the control, SO and HVF mice. Our results showed the beneficial effects of FO supplementation, as well as the harmful effects of trans FA, whose intensity can increase vulnerability to skin diseases.     

Characterisation of L-[3H]glutamate binding to fresh and frozen crude plasma membranes isolated from cerebral cortex of adult rats

Specific [3H]glutamate binding to fresh crude plasma membranes (CPMs) was compared with binding to frozen CPMs and the optimal conditions for the binding to frozen CPMs isolated from cerebral cortex of adult rats were determined. Freezing reduced [3H]glutamate binding (3.5-fold), and pre-incubation of previously frozen membranes followed by three washes increased binding (4.5-fold) when compared to fresh samples. CPMs washed once, pre-incubated at 37 degrees C and washed 3 times was adopted as the most adequate condition for the binding assay of frozen membranes. In a Cl(-)-containing medium, [3H]glutamate binding (Bmax=97.9 pmol/mg, Kd=349.68 nM) to this frozen CPM preparation was significantly displaced by excess quisqualic acid (QA) (65%), L-2-amino-4-phosphonobutyric acid (L-AP4) (35%), trans-1-aminocyclopentane-1,3-dicarboxylate (1S,3R-ACPD) (25%) and alfa-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) (25%). In a Cl(-)-free medium, binding (Bmax=44.14 pmol/mg, 311 nM) was significantly displaced by QA (45%), L-AP4 (25%), ACPD (25%), AMPA (25%), kainic acid (20%) and N-methyl-D-aspartate (15%).     

Saliva Proteomics as Fluid Signature of Inflammatory and Immune-Mediated Skin Diseases

Saliva is easy to access, non-invasive and a useful source of information useful for the diagnosis of serval inflammatory and immune-mediated diseases. Following the advent of genomic technologies and -omic research, studies based on saliva testing have rapidly increased and human salivary proteome has been partially characterized. As a proteomic protocol to analyze the whole saliva proteome is not currently available, the most common aim of the proteomic analysis is to discriminate between physiological and pathological conditions. The salivary proteome has been initially investigated in several diseases: oral squamous cell carcinoma and oral leukoplakia, chronic graft-versus-host disease, and Sjögren’s syndrome. Otherwise, salivary proteomics studies in the dermatological field are still in the initial phase, thus the aim of this review is to collect the best research evidence on the role of saliva proteomics analysis in immune-mediated skin diseases to understand the direction of research in this field. The results of PRISMA analysis reported herein suggest that human saliva analysis could provide significant data for the diagnosis and prognosis of several immune-mediated and inflammatory skin diseases in the next future.     

Lipid stability during the frozen storage of fillets from silver catfish exposed in vivo to the essential oil of Lippia alba (Mill.) NE Brown

BACKGROUND: Lippia alba is effective in sedating and reducing stress to fish during transportation. Because some in vitro studies have demonstrated the antioxidant activity of L. alba, we hypothesized that its use in vivo could result in antioxidant effects post mortem. Therefore, in this study we evaluated whether the essential oil of L. alba (EO) used as sedative for fish transport would increase the lipid stability of fillets from silver catfish during frozen storage. RESULTS: The exposure to the EO in vivo did not affect conjugated diene values. However, EO (30 and 40 µL L^?1) delayed the peak formation of peroxides (from the third to the sixth month of storage) and thiobarbituric reactive substances (from the ninth to the twelfth month of storage) when compared to control fillets. After exposure to 40 µL L^?1 EO the free fatty acid content was higher than for control at the start of fillet storage, with no differences among groups thereafter. CONCLUSION: The essential oil of L. alba used as sedative in the water to transport silver catfish can delay lipid oxidation of fillets during frozen storage. Thus L. alba may be a promising source of natural active compounds for use in aquaculture and the food industry. © 2012 Society of Chemical Industry     

Catabolism of Ap4A and Ap5A by rat brain synaptosomes

Adenosine 5',5"'-P1,P4-tetraphosphate (Ap4A) and adenosine 5',5"'-P1,P5-pentaphosphate (Ap5A) are stored in and released from rat brain synaptic terminals. In the present study we investigated the hydrolysis of dinucleotides (Ap4A and Ap5A) in synaptosomes from the cerebral cortex of adult rats. Ap4A and Ap5A, but not Ap3A, were hydrolyzed at pH 7.5 in the presence of 20 mM Tris/HCl, 2.0 mM MgCl2, 10 mM glucose and 225 mM sucrose at 37 degrees C. The disappearance of the substrates measured by FPLC on a mono-Q HR column was both time and protein dependent. Since synaptosome integrity was at least 90% at the end of the assay, hydrolysis probably occurred by the action of an ecto-enzyme. Extracellular actions of adenine dinucleotides at central nervous system terminate due to the existence of ecto-nucleotidases which specifically cleave these dinucleotides. These enzymes in association with an ATP diphosphohydrolase and a 5'-nucleotidase are able to promote the complete hydrolysis of dinucleotides to adenosine in the synaptic cleft.     

delta-Aminolevulinate dehydratase inhibition by 2,3-dimercaptopropanol is mediated by chelation of zinc from a site involved in maintaining cysteinyl residues in a reduced state

Phospholipase A2 (PLA2) hydrolyses membrane phospholipids (PL) and it may release arachidonic acid (AA)--the precursor of eicosanoids--from the sn-2 position. PLA2 and metabolites of its catalytic activity participate in many processes in the organism: metabolism of lipids, inflammation and immune reactions, membrane and tissue reparation, proliferation, and others. PLA2 as also an important element in the signal transduction. In the present article, PLA2 is characterised, classified into several types, and its mechanism of action together with its possible role in the disease processes are described. The attention is aimed at two forms of PLA2: The secretory (PLA2) of the type II which is associated with the inflammation injury, and the cytosolic PLA2 which is the main catalyst in the liberation of AA and which participates in the signal transduction. Other forms of PLA2 has been also described.