Combination of Hydrolysable Tannins and Zinc Oxide on Enterocyte Functionality: In Vitro Insights.

The management of gastrointestinal disease in animals represents a significant challenge in veterinary and zootechnic practice. Traditionally, acute symptoms have been treated with antibiotics and high doses of zinc oxide (ZnO). However, concerns have been raised regarding the potential for microbial resistance and ecological detriment due to the excessive application of this compound. These concerns highlight the urgency of minimizing the use of ZnO and exploring sustainable nutritional solutions. Hydrolysable tannins (HTs), which are known for their role in traditional medicine for acute gastrointestinal issues, have emerged as a promising alternative. This study examined the combined effect of food-grade HTs and subtherapeutic ZnO concentration on relevant biological functions of Caco-2 cells, a widely used model of the intestinal epithelial barrier. We found that, when used together, ZnO and HTs (ZnO/HTs) enhanced tissue repair and improved epithelial barrier function, normalizing the expression and functional organization of tight junction proteins. Finally, the ZnO/HTs combination strengthened enterocytes' defense against oxidative stress induced by inflammation stimuli. In conclusion, combining ZnO and HTs may offer a suitable and practical approach for decreasing ZnO levels in veterinary nutritional applications.

Protective effects of fatty acid amide hydrolase inhibition in UVB-activated microglia.

Neuroinflammation is a hallmark of several neurodegenerative disorders that has been extensively studied in recent years. Microglia, the primary immune cells of the central nervous system (CNS), are key players in this physiological process, demonstrating a remarkable adaptability in responding to various stimuli in the eye and the brain. Within the complex network of neuroinflammatory signals, the fatty acid N-ethanolamines, in particular N-arachidonylethanolamine (anandamide, AEA), emerged as crucial regulators of microglial activity under both physiological and pathological states. In this study, we interrogated for the first time the impact of the signaling of these bioactive lipids on microglial cell responses to a sub-lethal acute UVB radiation, a physical stressor responsible of microglia reactivity in either the retina or the brain. To this end, we developed an in vitro model using mouse microglial BV-2 cells. Upon 24 h of UVB exposure, BV-2 cells showed elevated oxidative stress markers and, cyclooxygenase (COX-2) expression, enhanced phagocytic and chemotactic activities, along with an altered immune profiling. Notably, UVB exposure led to a selective increase in expression and activity of fatty acid amide hydrolase (FAAH), the main enzyme responsible for degradation of fatty acid ethanolamides. Pharmacological FAAH inhibition via URB597 counteracted the effects of UVB exposure, decreasing tumor necrosis factor α (TNF-α) and nitric oxide (NO) release and reverting reactive oxidative species (ROS), interleukin-1ß (IL-1ß), and interleukin-10 (IL-10) levels to the control levels. Our findings support the potential of enhanced fatty acid amide signaling in mitigating UVB-induced cellular damage, paving the way to further exploration of these lipids in light-induced immune responses.

Comparative study on trace metal accumulation in the liver of two fish species (Torpedinidae): concentration-size relationship.

Metal concentrations (Hg, Cd, Pb, As, Cr, Cu, Zn and Ni) were measured in the liver of two fish, Torpedo nobiliana (electric ray) and Torpedo marmorata (marbled electric ray), from the Mediterranean Sea in order to comparatively investigate their current pollution status. Maximum mean levels of Hg were detected in electric ray (mean: 2.16µgg(-1) ww), while marbled electric ray accumulated especially Cd (mean: 0.06µgg(-1) ww), Cu (mean: 3.83µgg(-1) ww) and As (mean: 32.64µgg(-1) ww). The metal concentrations are similar to those reported in literature, except for Cd, As and Ni. Hg concentrations increased with increasing fish body length in both species, whilst no significant concentration-size relationship was found for other metals. To the best of our knowledge this is the first report providing information on trace metal levels and relationship between concentration and size of these cartilaginous fishes. Future studies on the concentrations and effects of environmental contaminants in various torpedinid species are surely needed.

Fluorimetric Assay of FAAH Activity.

Fatty acid amide hydrolase (FAAH) is the enzyme responsible for the degradation of anandamide (N-arachidonoylethanolamine, AEA) to arachidonic acid (AA) and ethanolamine. The method described here measures FAAH activity through the fluorometric arachidonoyl-7-amino-4-methyl-coumarin amide (AAMCA) substrate, which allows a simple and sensitive assay suitable for high-throughput screening tests. FAAH catalyzes the hydrolysis of AAMCA producing AA and the highly fluorescent compound 7-amino-4-methylcoumarin (AMC).

Contamination by polychlorinated biphenyls (PCBs) in striped dolphins (Stenella coeruleoalba) from the Southeastern Mediterranean Sea.

Concentrations of polychlorinated biphenyls (PCBs) including dioxin-like PCBs (non-ortho, PCB 77, PCB 126, and PCB 169 and mono-ortho, PCB 105, PCB 118, and PCB 156) were measured in different organs and tissues (melon, blubber, liver, kidney, lung, heart, and muscle tissue) of striped dolphins (Stenella coeruleoalba) from the Eastern Mediterranean Sea (Adriatic Sea). The mean highest levels were in blubber and melon, followed by liver, kidney, lung, heart, and muscle tissue. PCB profiles were similar in all tissues and organs being dominated by the higher chlorinated homologues (hexa-CBs, 55.8-62.1%; penta-CBs, 15.4-20.0%; and hepta-CB PCB 180, 12.7-16.5%). Major PCBs in all tissues were congeners 138 and 153 collectively accounting for 50.6-58.3% of the total PCB concentrations, followed by PCB 101, 105, 118, and 180 constituting from 27.0% to 31.0%. PCB levels were higher in adult males than in adult females. The estimated 2,3,7,8-TCDD toxic equivalents of non- and mono-ortho PCBs were much higher than the threshold level above which adverse effects have been observed in other marine mammals species, suggesting that striped dolphins in this region are at risk for toxic effects.

Cellular Prion Protein Expression in the Brain Tissue from Brucella ceti-Infected Striped Dolphins (Stenella coeruleoalba).

Brucella ceti, a zoonotic pathogen of major concern to cetacean health and conservation, is responsible for severe meningo-encephalitic/myelitic lesions in striped dolphins (Stenella coeruleoalba), often leading to their stranding and death. This study investigated, for the first time, the cellular prion protein (PrPc) expression in the brain tissue from B. ceti-infected, neurobrucellosis-affected striped dolphins. Seven B. ceti-infected, neurobrucellosis-affected striped dolphins, found stranded along the Italian coastline (6) and in the Canary Islands (1), were investigated, along with five B. ceti-uninfected striped dolphins from the coast of Italy, carrying no brain lesions, which served as negative controls. Western Blot (WB) and immunohistochemistry (IHC) with an anti-PrP murine monoclonal antibody were carried out on the brain parenchyma of these dolphins. While PrPc IHC yielded inconclusive results, a clear-cut PrPc expression of different intensity was found by means of WB analyses in the brain tissue of all the seven herein investigated, B. ceti-infected and neurobrucellosis-affected cetacean specimens, with two dolphins stranded along the Italian coastline and one dolphin beached in Canary Islands also exhibiting a statistically significant increase in cerebral PrPc expression as compared to the five Brucella spp.-negative control specimens. The significantly increased PrPc expression found in three out of seven B. ceti-infected, neurobrucellosis-affected striped dolphins does not allow us to draw any firm conclusion(s) about the putative role of PrPc as a host cell receptor for B. ceti. Should this be the case, an upregulation of PrPc mRNA in the brain tissue of neurobrucellosis-affected striped dolphins could be hypothesized during the different stages of B. ceti infection, as previously shown in murine bone marrow cells challenged with Escherichia coli. Noteworthy, the inflammatory infiltrates seen in the brain and in the cervico-thoracic spinal cord segments from the herein investigated, B. ceti-infected and neurobrucellosis-affected striped dolphins were densely populated by macrophage/histiocyte cells, often harboring Brucella spp. antigen in their cytoplasm, similarly to what was reported in macrophages from mice experimentally challenged with B. abortus. Notwithstanding the above, much more work is needed in order to properly assess the role of PrPc, if any, as a host cell receptor for B. ceti in striped dolphins.

Immunohistochemical investigations on Brucella ceti-infected, neurobrucellosis-affected striped dolphins (Stenella coeruleoalba).

Bacteria of the genus Brucella cause brucellosis, an infectious disease common to humans as well as to terrestrial and aquatic mammals. Since 1994 several cases of Brucella spp. infection have been reported in marine mammals worldwide. While sero-epidemiological data suggest that Brucella spp. infection is widespread globally, detecting Brucella spp.-associated antigens by immunohistochemistry (IHC) in tissues from infected animals is often troublesome. The present study was aimed at investigating, by means of IHC based upon the utilization of an anti-Brucella LPS monoclonal antibody (MAb), the central nervous system (CNS) immunoreactivity shown by B. ceti-infected, neurobrucellosis-affected striped dolphins. The aforementioned MAb, previously characterized by means of ELISA and Western Blotting techniques, was able to immunohistochemically detect smooth brucellae both within the CNS from B. ceti-infected striped dolphins and within a range of tissues from Brucella spp.-infected domestic ruminants. In conclusion, the results of the present study are of relevance both from the B. ceti infection's diagnostic and pathogenetic standpoints.

Relationship between total mercury concentration and fish size in two pelagic fish species: implications for consumer health.

Total mercury concentrations were determined in different size classes of two pelagic fish species of great commercial importance, horse mackerel (Trachurus trachurus) and Mediterranean horse mackerel (Trachurus mediterraneus), to evaluate the relationship between total mercury concentration and fish size and to determine whether any differences might affect the quantitative assessment of mercury exposure for consumers. Mercury concentrations in horse mackerel and in Mediterranean horse mackerel were between 0.16 and 2.41 microg g(-1) of weight wet (mean, 0.68 microg g(-1)) and between 0.09 and 1.62 microg g(-1) (mean, 0.51 microg g(-1)), respectively. The regression curves revealed a significant relationship between mercury concentration and fish size (length and weight) for both species. Concentrations exceeding the proposed limit for human consumption were observed in 33.3% of the samples of both species and were associated with larger specimens. The consumption of the larger specimens could lead to an increase in mercury exposure for consumers. Estimated weekly intakes, calculated on the basis of concentrations relative to each size class, revealed a high exposure associated with the consumption of fish larger than 30 cm (horse mackerel, 11.63 to 20.16 microg/kg of body weight; Mediterranean horse mackerel, 5.86 to 13.55 microg/kg of body weight). An understanding of the factors leading to an increase in mercury exposure can help consumers make informed decisions about eating fish.

Evaluation of trace metal and polychlorinated biphenyl levels in tea brands of different origin commercialized in Italy.

The objectives of this study were to investigate the trace element (Hg, Cd, Pb, Cu, Zn, Ni, Fe, Cr and Se) and polychlorinated biphenyl (PCBs) content of several commercially available brands of green and black tea marketed in Italy. The concentrations these chemicals were found to be variable and largely dependent upon the type and brand of analysed tea. The most abundant element among the essential elements was Fe, followed by Zn, Cu, Se, Ni and Cr, whereas Pb was the predominant among the tested nonessential elements followed by Hg and Cd. Assessment based on several available guidelines showed that element content were low, except for Hg and Ni. The PCBs concentrations were generally low, with a homologue profile dominated by low-chlorinated congeners, namely three- and tetra-PCBs accounting for more than 60% of the total residue. Apart from trace elements, this is the first study documenting in detail the concentrations and congener distribution of PCBs in tea samples of different origin.

Health risk of coplanar polychlorinated biphenyl congeners in edible fish from the Mediterranean Sea.

The muscle tissue of different species of fish was analyzed for polychlorinated biphenyls (PCBs) in order to estimate the daily intake of these contaminants. Contamination levels among the different species varied between 108 and 678 ng/g lipid weight. Isomer-specific analysis in the muscle tissue of the various fish revealed a profile dominated by hexa- and pentachlorobiphenyls, followed by hepta- and tetrachlorobiphenyls. Other congeners, including those with fewer than four or more than nine chlorine atoms, were below the instrumental limit of detection in all samples. PCB dietary intake was below the range of 1 to 4 pg 2,3,7,8-tetrachloro-p-dioxin equivalents per kg body weight per day set by the World Health Organization. From a public health point of view, there is no indication of important risks associated with the consumption of these seafoods.

PCBs and PCDD/PCDFs in fishery products: occurrence, congener profile and compliance with European Union legislation.

PCB and PCDD/F concentrations and congener specific profiles were determined in seafood (fish, cephalopods, crustaceans) purchased in supermarkets in Southern Italy. The results intended to assess and compare the levels and congener profiles among the various organisms and evaluate whether the pollutant concentrations exceed the maximum permitted levels according to European Union guidelines. Contaminant accumulation followed the order PCBs > PCDFs > PCDDs in all samples. Species-specific bioaccumulation of contaminants and differences in PCB and PCDD/F profiles among the three different groups of seafood were noted. Pollutant concentrations, given as sum of WHO-TEQ, in all samples (fish: 0.28 pg g(-1) wet wt; cephalopods: 0.01 pg g(-1) wet wt; crustaceans: 0.03 pg g(-1) wet wt) did not exceed the maximum limits for human consumption. Also the sum of six indicator PCBs (fish: 0.07-16.7 ng g(-1) wet weight; cephalopods: 0.05-0.21 ng g(-1) wet weight; crustaceans: 0.07-0.57 ng g(-1) wet weight) was below the level prescribed by the legislation in all the species tested. The estimated dietary intakes (fish: 0.52 TEQs/kg bw/week; cephalopods: 0.01 TEQs/kg bw/week; crustaceans: 0.02 TEQs/kg bw/week) were below the tolerable weekly intake (TWI) proposed by the European Commission. Therefore, the samples analyzed can be considered safe considering the levels obtained and the in-force legislation.

The expression and activity of 5-LOX in the large intestine of horses harbouring encysted cyathostomin larvae.

Leukotrienes are products of the arachidonic acid metabolism and act as potent inflammatory mediators modulating the immune response and various physiological processes. This study evaluated the expression and activity of 5-lipoxygenase (5-LOX), the enzyme that catalyzes the first two steps in the biosynthesis of leukotrienes, in horses infected by larval cyathostomins. Tissue samples from dorsal and ventral colon, and from the cecum were collected from 16 horses slaughtered for human consumption. Samples were analyzed to estimate the burdens of encysted cyathostomin larvae and adult luminal stages, and then processed for the evaluation of biochemical parameters. No significant differences were found in the protein expression and enzymatic activity of 5-LOX between animals harbouring only adult parasites and negative horses. The protein expression and enzyme activity of 5-LOX were significantly higher in horses harbouring encysted larvae in comparison with horses free of encysted larvae. Although preliminary, these results indicate that 5-LOX is an important mediator in the course of horse cyathostominosis and further studies are warranted to unveil the possible role this enzyme plays in the pathogenesis of horse cyathostominosis, and its potential as a diagnostic marker.

Toxic metals (Hg, Cd, and Pb) in fishery products imported into Italy: suitability for human consumption.

Mercury, cadmium, and lead concentrations were determined in various fishery products (fishes, cephalopod molluscs, and crustaceans) imported into Italy from many European and non-European coastal countries. Considerable differences were found in the concentrations of these metals among the products tested. The highest mean Hg concentration was found in fishes (0.21 µg g(-1) wet weight), whereas cephalopods had the highest mean Cd concentration (0.35 µg g(-1) wet weight). Swordfish (0.80 µg g(-1) wet weight), longtail tuna (0.53 µg g(-1) wet weight), and thornback ray (0.52 µg g(-1) wet weight) had the highest concentrations of Hg, whereas maximum Cd concentrations were found in samples of common cuttlefish (0.85 µg g(-1) wet weight) and common octopus (0.64 µg g(-1) wet weight). The majority of the samples analyzed were in compliance with European Union legislation, except for a few cases. The calculated mean weekly intakes of Hg, Cd, and Pb through consumption of the fishery products tested were all below the legislated respective provisional tolerable weekly intakes. In general, the samples analyzed were considered safe to eat with regard to the metal concentrations found and the allowable intakes based on legislation. Nevertheless, the consumption of some species may be of significant importance for consumer health.

Vitamin E-related inhibition of monocyte 5-lipoxygenase and cardiovascular outcome in maintenance hemodialysis patients.

A daily supplement of vitamin E is recommended for the secondary prevention of cardiovascular events in end-stage renal disease patients on maintenance hemodialysis. Vitamin E has been entrusted with therapeutic properties against cardiovascular disease for more than 60 years. Several epidemiological studies and intervention trials have been performed with vitamin E, and some of them showed that it prevents atherosclerosis. For a long time, vitamin E was assumed to act by decreasing the oxidation of low-density lipoproteins, a key step in atherosclerosis initiation. However, at the cellular level vitamin E interferes with smooth muscle cell proliferation, platelet aggregation, monocyte adhesion, and oxidized low-density lipoproteins uptake and cytokine production, all reactions implied in the progression of atherosclerosis. Recent research points out that these effects may be not only the result of the antioxidant activity of vitamin E but also of its distinct molecular actions. These biological properties of vitamin E may allow to design better strategies for primary and secondary prevention of cardiovascular disease, with a potential exploitation of vitamin E supplements in primary and secondary prevention of major adverse cardiovascular events in all uremic patients. In this review, we also outline relevant patents on vitamin E and lipoxygenase inhibitors.

Relationships between paraoxon and 2-coumaranone hydrolytic activities in sera genotyped for PON1 Q192R polymorphism.

OBJECTIVES: To set-up a method for a direct evaluation in human serum of paraoxonase enzymatic activities, establishing a possible correlation with Q192R genotype polymorphism. DESIGN AND METHODS: 101 different human serum samples were genotyped for paraoxonase Q192R polymorphism by PCR restriction analysis, and evaluated spectrophotometrically with regard to paraoxon and 2-coumaranone hydrolytic activities. Both activities of paraoxonase were assayed, quantified through normalization by arylesterase activity, and compared with the data concerning Q/R genetic polymorphism. RESULTS: The mean normalized paraoxonase activity was found to be significantly higher in RR than in QQ human sera (3.99+/-0.6 versus 1.32+/-0.44; P<0.0001); instead, the 2-coumaranone hydrolysis showed an opposite trend (0.10+/-0.02 versus 0.23+/-0.04, in RR and QQ sera respectively; P<0.0001). CONCLUSIONS: These methods were successfully applied to the whole serum, suggesting a possible use of this approach for a clinically relevant phenotypic characterization.