Paronychia argentea Lam. protects renal endothelial cells against oxidative injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Paronychia argentea Lam. (Arabic tea), a species spontaneously growing in the Mediterranean area, has been used in folk medicine for renal diseases. AIM OF THE STUDY: To assess the antioxidant and protective potentials of different extracts from P. argentea in the renal endothelial NRK-52E cell line by several in vitro models, including a H2O2-induced oxidative stress model. MATERIAL AND METHODS: Aerial parts of P. argentea were collected in Algeria and ethanolic, chloroform and aqueous-chloroform extracts were obtained from dried plant. The antioxidant capacity was first evaluated by the Oxygen Radical Absorbance Capacity (ORAC) and the free radical scavenging activity (DPPH) methods. Cellular viability was assessed by MTT method assay after 24 h pretreatment with each extract concentration in order to measure protection from H2O2 in NRK-52E cells. Furthermore, the intracellular ROS formation (DCFH-DA method), was determined. RESULTS: P. argentea showed in vitro antioxidant activity as evidenced by the ORAC and DPPH assays. No cell toxicity was observed for concentrations ranging from 0.1 to 100 µg/mL of each extract. These extracts also exerted a protective effect on renal endothelial cells simultaneously treated with 1 mM H2O2. Chemical composition for the aqueous-chloroform extract was assessed by HPLC, as it showed the strongest antioxidant ability, revealing three quercetin derivatives as the main phenolic compounds. CONCLUSION: P. argentea is endorsed with antioxidant activity and protects renal endothelial cells against oxidative damage which indicate this plant constitutes a potential treatment for renal diseases.

Effects of Fiber Purified Extract of Carob Fruit on Fat Digestion and Postprandial Lipemia in Healthy Rats.

Increased postprandial lipemia is a cardiovascular disease (CVD) risk factor. Carob fruit extract (CFE) contains condensed tannins, and their intake has been inversely related to CVD. The objective was to evaluate the in vitro pancreatic lipase activity in the presence of CFE and the in vivo effect of CFE on postprandial lipemia of healthy Wistar rats in acute and subchronic digestibility studies and to relate it with changes in fat digestion and absorption. CFE significantly reduced pancreatic lipase activity. A peak delay and a dose-dependent decrease in plasma triglyceride and cholesterol areas under the curve were observed, effects that increased after the subchronic treatment. The levels of nondigested, nonabsorbed triglycerides of the remaining intestinal lumen fat were significantly higher in the maximum dose of CFE administrated versus the control ( P < 0.05). This study demonstrates for the first time the hypolipemic properties of CFE from the first administration, modifying postprandial lipemia by reducing the extents of fat digestion and absorption.

Acanthocotyle gurgesiella n. sp. (Monogenea: Acanthocotylidae) from the deep-sea skate Gurgesiella furvescens (Rajidae) in the south-eastern Pacific.

Little is known about the diversity of parasites of the deep-sea fish of the world's oceans. Here, a new species of monogenean parasite of the deep-sea skate Gurgesiella furvescens is described. Specimens of parasites were obtained from the skin of two specimens of the dusky finless skate, G. furvescens (Rajidae), in the vicinity of Valparaiso (33°S, 72°W), central Chile, from midwater trawl fishing at depths of 350-450 m. Both morphological and molecular analyses were conducted to provide a full description of the new species, named Acanthocotyle gurgesiella. For the molecular analyses, nuclear large subunit (LSU) rDNA and the mitochondrial gene cytochrome c oxidase 1 (COI) were used. From the morphological analysis and a comparison with the known species of the genus, A. gurgesiella can be identified by a combination of morphological characteristics, including the number of testes, number of radial rows of sclerites in the pseudohaptor, aperture of the genital pore and shape of the vitelline follicles. The results from the DNA analysis indicated that A. gurgesiella has a genetic divergence of 3.2-3.7% (LSU rDNA gene) from A. urolophi, the only congener species for which molecular data are available.

Fiber purified extracts of carob fruit decrease carbohydrate absorption.

The postprandial state plays a central role in the development and setting of chronic diseases. Condensed tannins (CT) are polyphenols with a known ability to modify carbohydrate digestion and absorption. The high concentration of CT in the pulp of carob fruit suggests a potential antidiabetic effect. The aim of this work was to analyze the in vitro and in vivo effects of carob fruit extract (CFE) on the digestion and absorption of carbohydrates. α-Glucosidase activity and glucose diffusion were tested in vitro using 0.5, 1, 2 and 5 mg mL-1 CFE concentrations. Two in vivo absorption studies, acute and subchronic, were carried out in four groups of 6 two-month-old male Wistar rats (control and CFE 25, 50 and 150 mg per kg b.w.), administering 1 mL of olive oil and 0.5 g per kg b.w. of glucose solution by oral gavage. CFE significantly inhibited α-glucosidase activity, through a competitive mechanism, from 1 mg mL-1, and also reduced glucose diffusion in a dose-dependent manner. In the acute study, CFE (50 and 150 mg per kg b.w.) significantly reduced the area under the curve (AUC) of blood glucose. Subchronic CFE administration induced further AUC decreases; and CFE at 150 mg per kg b.w. reduced sodium-glucose-linked transporter-1 (SGLT1) levels in the duodenum. This study demonstrates the hypoglycemic properties of CFE, highlighting its potential role as a suitable nutritional strategy in diabetic patients.

Could post-weaning dietary chia seed mitigate the development of dyslipidemia, liver steatosis and altered glucose homeostasis in offspring exposed to a sucrose-rich diet from utero to adulthood?

The present work analyzes the effects of dietary chia seeds during postnatal life in offspring exposed to a sucrose-rich diet (SRD) from utero to adulthood. At weaning, chia seed (rich in α-linolenic acid) replaced corn oil (rich in linoleic acid) in the SRD. At 150 days of offspring life, anthropometrical parameters, blood pressure, plasma metabolites, hepatic lipid metabolism and glucose homeostasis were analyzed. Results showed that chia was able to prevent the development of hypertension, liver steatosis, hypertriglyceridemia and hypercholesterolemia. Normal triacylglycerol secretion and triacylglycerol clearance were accompanied by an improvement of de novo hepatic lipogenic and carnitine-palmitoyl transferase-1 enzymatic activities, associated with an accretion of n-3 polyunsaturated fatty acids in the total composition of liver homogenate. Glucose homeostasis and plasma free fatty acid levels were improved while visceral adiposity was slightly decreased. These results confirm that the incorporation of chia seed in the diet in postnatal life may provide a viable therapeutic option for preventing/mitigating adverse outcomes induced by an SRD from utero to adulthood.

Helminth parasites of South American fishes: current status and characterization as a model for studies of biodiversity.

The South American subcontinent supports one of the world's most diverse and commercially very important ichthyofauna. In this context, the study of South American fish parasites is of increased relevance in understanding their key roles in ecosystems, regulating the abundance or density of host populations, stabilizing food webs and structuring host communities. It is hard to estimate the number of fish parasites in South America. The number of fish species studied for parasites is still low (less than 10%), although the total number of host-parasite associations (HPAs) found in the present study was 3971. Monogeneans, with 835 species (1123 HPAs, 28.5%), and trematodes, with 662 species (1127 HPAs, 30.9%), are the more diverse groups. Data gathered from the literature are useful to roughly estimate species richness of helminths from South American fish, even though there are some associated problems: the reliability of information depends on accurate species identification; the lack of knowledge about life cycles; the increasing number of discoveries of cryptic species and the geographically biased number of studies. Therefore, the closest true estimations of species diversity and distribution will rely on further studies combining both molecular and morphological approaches with ecological data such as host specificity, geographical distribution and life-cycle data. Research on biodiversity of fish parasites in South America is influenced by problems such as funding, taxonomic impediments and dispersion of research groups. Increasing collaboration, interchange and research networks in the context of globalization will enable a promising future for fish parasitology in South America.

Metazoan parasites of Brama australis from southern Chile: a tool for stock discrimination?

The metazoan parasites of 403 specimens of the southern ray's bream Brama australis from three localities in southern Chile (Lebu 36° 70' S; 73° 40' W, Calbuco 41° 50' S; 73° 08' W and Punta Arenas 53° 10' S; 70° 50' W) were recorded. More than 23 400 parasite specimens belonging to 12 taxa were registered. Metazoan parasites were dominated by the copepod Hatschekia conifera, constituting 97% of the total number of parasites; the larval cestode Hepatoxylon trichiuri was the second most important parasite (2·1% of the total number of parasites). The remaining 10 species constituted <1% of the metazoan parasites. Parasitological evidence, based on univariate and multivariate analysis, does not support the existence of discrete stocks in the studied zone.

Mechanisms underlying the beneficial effect of soy protein in improving the metabolic abnormalities in the liver and skeletal muscle of dyslipemic insulin resistant rats.

PURPOSE: The present study analyzes the effect of the replacement of dietary casein by soy protein on the mechanisms underlying dyslipidemia, liver steatosis and altered glucose and lipid metabolism in the skeletal muscle which developed in rats fed long-term a sucrose-rich diet (SRD). METHODS: Wistar rats were fed a SRD for 4 months. From months 4 to 8, half the animals continued with the SRD, and the other half were fed a SRD in which the source of protein casein was replaced by soy. The control group received a diet with cornstarch as source of carbohydrate. RESULTS: Compared to SRD-fed animals, the rats fed soy showed: A--in the liver: reduction of triglyceride and cholesterol storage and decreased steatosis; normalization of mature forms of the protein mass levels of SREBP-1 and the activities of lipogenic enzymes, while the protein mass level of PPAR-α and fatty acid oxidase activity increased. B-in the gastrocnemius muscle: normalization of the enhanced lipid storage and the altered glucose oxidation, improving glucose phosphorylation; decreasing protein mass level of nPKCθ in the membrane fraction; reversion of the impaired insulin-stimulated glucose transporter Glut-4, and glucose-6-phosphate and glycogen concentrations. Besides, dyslipidemia and glucose homeostasis returned to control values. CONCLUSIONS: This study provides new information concerning some key mechanisms related to the effect of dietary soy on hepatic lipid metabolism and insulin action in the skeletal muscle in the presence of pre-existing dyslipidemia and insulin resistance induced by a SRD.

Two new species of Encotyllabe (Monogenea: Capsalidae) based on morphometric and molecular evidence: parasites of two inshore fish species of northern Chile.

Two new species, Encotyllabe cheilodactyli and Encotyllabe antofagastensis (Monogenea: Capsalidae), have been found in the pharyngeal plates of Cheilodactylus variegatus and Anisotremus scapularis, respectively, in northern Chile (23°38'N, 70°24'W). Descriptions of the new species were based on morphometric and molecular evidence. Both species differ from previously described species of the genus by a combination of characteristics, including the size and relative position of the testes and the shapes of the major and small hamulus. In addition, E. cheilodactyli is unique among the known species of Encotyllabe due to the testes crossing the equatorial line of the body proper, and E. antofagastensis is unique among the known species of Encotyllabe due to its penis structure and anatomy leading to the genital pore. The analysis of the cytochrome c oxidase I gene as well as morphometric analyses demonstrated that the specimens belonged to 2 different species.

Does a facultative precocious life cycle predispose the marine trematode Proctoeces cf. lintoni to inbreeding and genetic differentiation among host species?

Intraspecific variability in parasite life cycle complexity (number of hosts and species of hosts in the life cycle) may have an impact how parasite genetic variation is partitioned among individual parasites, host individuals or host species within a given area. Among digenean trematodes, a three-host life cycle is common. However, a few species are precocious and may reach sexual maturity in what is typically regarded as the second intermediate host. The objective of this study was to determine whether a precocious life cycle predisposes digeneans to possible inbreeding or genetic subdivision among host species. As a study system, we used the digenean Proctoeces cf. lintoni whose metacercariae precociously mature (facultative) without a cyst wall in the gonads of multiple sympatric species of keyhole limpets (Fissurella spp.), typically regarded as the second intermediate hosts. Genotyped parasites were collected from four species of limpets and the clingfish Sicyases sanguineus, the third and final host where sexual maturity occurs. We found very high microsatellite diversity, Hardy-Weinberg equilibrium over all genotyped individuals, and little to no genetic structuring among parasites collected from the different host species. The fact that metacercariae do not encyst in the keyhole limpets, coupled with the high mixing potential of an aquatic environment, likely promote panmixia in local populations of P. cf. lintoni.

Dietary Salba (Salvia hispanica L) seed rich in α-linolenic acid improves adipose tissue dysfunction and the altered skeletal muscle glucose and lipid metabolism in dyslipidemic insulin-resistant rats.

This work reports the effect of dietary Salba (chia) seed rich in n-3 α-linolenic acid on the morphological and metabolic aspects involved in adipose tissue dysfunction and the mechanisms underlying the impaired glucose and lipid metabolism in the skeletal muscle of rats fed a sucrose-rich diet (SRD). Rats were fed a SRD for 3 months. Thereafter, half the rats continued with SRD while in the other half, corn oil (CO) was replaced by chia seed for 3 months (SRD+chia). In control group, corn starch replaced sucrose. The replacement of CO by chia seed in the SRD reduced adipocyte hypertrophy, cell volume and size distribution, improved lipogenic enzyme activities, lipolysis and the anti-lipolytic action of insulin. In the skeletal muscle lipid storage, glucose phosphorylation and oxidation were normalized. Chia seed reversed the impaired insulin stimulated glycogen synthase activity, glycogen, glucose-6-phosphate and GLUT-4 protein levels as well as insulin resistance and dyslipidemia.

Is Anisakis simplex s.l. a biological marker for stock identification of Strangomera bentincki from Chile?

The absence of the larval Anisakis simplex s. l. in samples from central Chile but heavy infections in fish from southern localities suggest the existence of two stocks of Strangomera bentincki, one closely associated with the central coast of Chile (landing port San Antonio and Talcahuano, c. 33° 30' S-36° 40' S) and the other associated with southern Chile (landing port Puerto Montt, c. 41° 30' S). Results confirm the usefulness of metazoan parasites, like A. simplex s. l. as biological markers.

Endoparasite fauna of five Gadiformes fish species from the coast of Chile: host ecology versus phylogeny.

The aims of the present study were to compare, using multivariate analyses, the degree of similarity of the endoparasite fauna of five fish species belonging to the order Gadiformes: Merluccius gayi, Merluccius australis, Macruronus magellanicus (Gadoidei) and Micromesistius australis and Nezumia pulchella (Macrouroidei), from the southern and central Chilean coast, and to evaluate whether the composition of the endoparasite fauna was determined by phylogenetic or ecological relationships. We employed our database of Merluccius australis, M. magellanicus and Micromesistius australis, which was complemented with published information for M. magellanicus, Merluccius australis, Micromesistius australis, M. gayi and N. pulchella. A higher number of endoparasite species was recorded for Merluccius australis, Micromesistius australis and M. magellanicus, namely Anisakis sp. and Hepatoxylon trichiuri, which is the most prevalent parasite among these hosts. Aporocotyle wilhelmi and Hysterothylacium sp. were detected only in M. gayi, whereas Lepidapedon sp. was found exclusively in N. pulchella. These results suggest that fish ecology rather than host phylogeny was the most important factor for the determination of similarity in parasite composition. This result could be explained by the similar trophic patterns of hosts and by the predominance of generalist larval species among these fish parasite communities.

Permanent genetic resources added to Molecular Ecology Resources Database 1 February 2011-31 March 2011.

This article documents the addition of 111 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Acipenser oxyrinchus desotoi, Anopheles nuneztovari sensu lato, Asellus aquaticus, Calopteryx splendens, Calopteryx virgo, Centaurea aspera, Centaurea seridis, Chilina dombeyana, Proctoeces cf. lintoni and Pyrenophora teres f. teres.

Metazoan parasites as biological tags for stock discrimination of whitemouth croaker Micropogonias furnieri.

Examination of 248 adult specimens of whitemouth croaker Micropogonias furnieri from five localities along the Brazilian coast revealed 8735 parasites belonging to 41 metazoan species. Samples from Ceará to Bahia and Rio de Janeiro to Santa Catarina showed a high level of correct assignation (92 and 87%, respectively) and cross assignation (i.e. almost all specimens misidentified in Ceará were assigned to Bahia and almost all specimens misidentified in Bahia were classified as Ceará), so samples were pooled in the northern and south-eastern samples, and Rio Grande do Sul was considered a southern area. Eight parasite species were characteristic of the northern localities, five species were found just in the area associated with south-eastern localities and two species were characteristic of the southern area providing first evidence of stock discreteness. The multivariate discriminant analysis successfully discriminated three groups of localities associated with three stocks of M. furnieri in Brazil: a northern stock associated with Ceará and Bahia, a south-eastern stock related to Rio de Janeiro and Santa Catarina and a southern stock in the area of Rio Grande do Sul, which could be considered as the northern limit of the stock associated with the Common Fishing Zone of Uruguay and Argentina.

Molecular evidence confirms that Proctoeces humboldti and Proctoeces chilensis (Digenea: Fellodistomidae) are the same species.

Two species of Proctoeces Odhner, 1911 have been described in marine organisms from Chile: P. humboldti George-Nascimento & Quiroga (1983), parasitizing the gonads of keyhole limpets (Fissurella spp.), and P. chilensis Oliva (1984), an intestinal parasite of Sicyases sanguineus (Teleostei); both species were subsequently considered as P. lintoni Siddiqi & Cable (1960). To assist in the resolution of the taxonomic identification of Proctoeces species in marine organisms from Chile, phylogenetic studies using DNA sequences from the V4 region of the SSU rRNA gene were performed. Several specimens of P. lintoni were isolated from keyhole limpets (Fissurella spp.) and clingfish (S. sanguineus) from Bahia San Jorge (23°40'S) and Bahia Concepción (36°50'S). Phylogenetic analyses were conducted using three different approaches: a neighbour-joining (NJ), a maximum likelihood (ML) and Bayesian inference (BI). The phylogenetic analysis confirms that specimens of Proctoeces obtained from keyhole limpets and those specimens from the clingfish are in fact the same species. We prefer to consider our specimens as Proctoeces cf. lintoni, as the morphology of Proctoeces appears to be of doubtful value and genetic information about P. lintoni Siddiqi & Cable (1960) is not available. In addition, our results strongly suggest that there are at least three species in this genus.

Two new species of Choricotyle Van Beneden & Hesse (Monogenea: Diclidophoridae), parasites from Anisotremus scapularis and Isacia conceptionis (Haemulidae) from northern Chilean coast.

Two new species, Choricotyle scapularis and Choricotyle isaciencis (Monogenea: Diclidophoridae), are described from the gills of haemulid fishes, Anisotremus scapularis and Isacia conceptionis, respectively, in San Jorge Bay, northern Chile (23 degrees 42'S, 70 degrees 24'W); they are compared with the known species in the genus. Diagnostic characters for C. scapularis includes the number of hooks in the male copulatory organ, a developed isthmus, the shape of the haptor, and the number of testes. The diagnostic characters for C. isaciensis include the presence of an oval accesory sclerite in the clamps, the number of testes, the number of hooks in the male copulatory organ, and the relative clamp peduncle size.

Is the nestedness of metazoan parasite assemblages of marine fishes from the southeastern Pacific coast a pattern associated with the geographical distributional range of the host?

Nested structure is a pattern originally described in island biogeography to characterize how a set of species is distributed among a set of islands. In parasite communities, nestedness has been intensively studied among individual fish from a locality. However, nested patterns among parasite assemblages from different host populations (localities) have scarcely been investigated. We recorded the occurrence of parasites in 9 fish species widely distributed along the southeastern Pacific coast to determine whether the ecto- and endoparasite assemblages of marine fishes show a nested structure associated with host distributional range. Nestedness was tested using Brualdi-Sanderson index of discrepancy (BR); and 5 null models incorporated in a 'Nestedness' programme (Ulrich, 2006). The ecto- and endoparasite richness do not show similar patterns of latitudinal gradients among fish hosts, with 33-66% of analysed ectoparasite assemblages, and 25-75% of endoparasite assemblages showing nested structures through the host distributional range. For ectoparasites, species richness gradients and nested structure (when present) might be associated with decreased host densities or could reflect negative environmental conditions in the distributional border of the host species, whereas for endoparasites might be caused by geographical breaks of prey or changes in prey availability (intermediate hosts). The sampled extension of the distributional range of the host species, as well as the lack of specificity of some parasites, could influence the detection of nestedness.

Growth hormone modulates the degradative capacity of muscle nucleases but not of cathepsin D in post-weaning mice.

We determined whether recombinant human growth hormone (rhGH) administration might modulate the enzyme degradative capacity of the muscle lysosomal system and influence muscle growth. Muscle cathepsin D, acid RNase and DNase II activities are determined in the gastrocnemius muscle of rhGH-treated post-weaning female BALB/c mice. Linear regressions were used to analyze the relationships of each enzyme with their respective substrate. GH induced a depletion-recovery response of muscle growth through a mechanism which is similar to catch-up growth. In these conditions, cathepsin D activity decreased with age in all animals (GH: 40%; saline: 79%), showing a substantial developmental decline that could reflect changes in the rate of protein breakdown. However, the degradative capacity of cathepsin D was paradoxically unmodified in rhGH-mice compared with saline mice (according to the enzyme vs. substrate linear regression slope), in spite of the increase in enzyme activity elicited by GH. This suggests that the muscle protein breakdown is not increased by GH-treatment in post-weaning mice. The enhancement of muscle protein deposition as indicated by the augmented muscle cell size (protein:DNA ratio) of rhGH-mice (increased 178% from 25 to 50 days) vs. saline, can be attributed to a higher muscle K(RNA). In contrast, acid RNase and DNase II activities directly participate in muscle RNA and DNA degradation. Both nucleases were inhibited by GH treatment (a decrease of 48% and 63%, respectively, vs. saline at 50 days). The decrease in RNase activity suggests an inverse relation between the rate of protein synthesis (high) and acid RNase activity (low), leading to spare muscle RNA for synthesizing protein during catch-up growth. Also, low DNase II activity could contribute to inhibiting of muscle DNA degradation, facilitating muscle growth. Thus, GH seems to act as a direct modulator of the degradative capacity of skeletal muscle nucleases but not of cathepsin D, influencing DNA and RNA degradation during the depletion-recovery response to GH of gastrocnemius muscle in female post-weaning mice.

The modulator effect of GH on skeletal muscle lysosomal enzymes is dietary protein dependent.

OBJECTIVE: The purpose of this work is to determine whether changes in dietary protein level could alter the modulator effect that GH has on the muscle lysosomal system by influencing the hydrolytic activities of cathepsin D, acid RNase and DNase II and the participation of these enzymes in muscle growth. DESIGN: BALB/c female mice were fed a diet containing 20% (HP) or 12% (MP) protein ad libitum and were treated with either saline (s) or rhGH (GH) (74 ng/g) for 29 days. Body weight and feed intake were recorded daily. At 25, 30, 35, 40, 45 and 50 days of age, five mice from each group were slaughtered and nucleic acids and protein concentrations and cathepsin D, acid RNase and DNase II activities in gastrocnemius muscle were analysed. Correlation coefficients were used to analyse the links between the activity of each enzyme with its substrate. RESULTS: GH-treatment induced a depletion-recovery response in muscle growth through a compensatory mechanism. Changes in protein content, DNA and RNA concentrations were related to changes in lysosomal enzyme activities. Muscle cathepsin D activity in saline mice fell as the dietary protein concentration increased. GH-treatment reversed this effect by enhancing the proteolytic activity in muscle of well-fed mice and inhibiting it in mice fed a 12% protein diet. This inversion appears to be related to the different mechanism elicited by GH-treatment on skeletal muscle protein growth in each dietary group. An opposite trend was observed in muscle acid nuclease activities. Acid RNase and DNase II increased according to the dietary protein concentration, since a 12% protein diet induced a lower catabolism, especially on muscle DNA of saline mice. In contrast, GH-treatment decreased acid RNase and DNase II activities, but only in mice fed a 20% protein diet, perhaps leading to spare muscle RNA for protein synthesis, as well as to the inhibition of DNA degradation during catch-up growth. A lower dietary protein concentration appeared to reverse the GH protective effect on nucleic acids. CONCLUSIONS: GH seems to act as a dietary protein-dependent modulator of the skeletal muscle lysosomal enzyme activity. These lysosomal enzymes play a role during muscle growth in GH-treated post-weaning mice by modifying muscle protein and DNA and RNA degradation.