Phenolic constituents and antioxidant properties of Xanthosoma violaceum leaves

An extract of Xanthosoma violaceum leaves was subjected to a polyphenol profile determination, including total polyphenols, and antioxidant activity evaluation. Analysis of the extract resulted in the isolation of a new flavone C-glycoside, apigenin 6-C-beta-D-glucopyranosyl-8-C-beta-D-apiofuranoside (1), as well as known flavone C-glycosides, including vitexin (2), isovitexin (3), isovitexin 4'-O-rhamnopyranoside (4), apigenin 6-C-[beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside] (5), and apigenin 6,8-diC-beta-D-glucopyranoside (6). The antioxidant activity of the extract was assessed by means of two different in vitro tests: bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH test) and peroxidation induced by the water-soluble radical initiator 2,2'-azobis(2-amidinopropane) hydrochloride, on mixed dipalmitoylphosphatidylcholine/linoleic acid unilamellar vesicles (LP-LUV test). In both tests used, the extract and a fraction II showed a significant antioxidant/free-radical scavenging effect (fraction II, EC(50) = 11.6 microg/mL) in comparison to alpha-tocopherol (EC(50) = 10.1 microg/mL).     

Relationship between oxidative stress and the success of artificial insemination in dairy cows in a pasture-based system

This study was designed to evaluate whether the outcome of artificial insemination (AI) was affected by the metabolic and oxidative status of dairy cows. Seventy-nine inseminations in 40 cows, were classified, on the basis of blood progesterone (P4) and pregnancy-associated glycoprotein (PAG) concentrations and clinical confirmation of pregnancy into, three categories: (1) positive (AI+, resulted in pregnancy, n=26; 33%), (2) negative (AI-, did not result in pregnancy, n=49; 62%), and (3) embryonic mortality (EM, n=4; 5%). Reactive oxygen metabolites, biological antioxidant potential, oxidative stress index, body condition score, glucose, total proteins, albumin, urea, non-esterified fatty acids (NEFAs), cholesterol, triglycerides, haptoglobin and advanced oxidative protein products (AOPPs) were measured on the day of AI (day 0), and 30 and 42days later. Cows with EM had lower BCS scores (2.5) than AI+ (2.8) and AI- (2.9) cows (P<0.05). During the post-partum period, body condition score (BCS) increased and NEFAs decreased (P<0.05) suggesting a recovery from the negative energy balance (NEB). The only significant differences found were that the mean concentration of AOPPs was higher and that of albumin lower in EM cows than in AI+ and AI- (P<0.05) animals. Plasma concentration of reactive oxygen metabolites and biological antioxidant potential were not related to AI outcome. Further studies are required to confirm this finding and to clarify the role of oxidative status on cows' fertility.     

Isolation of new pregnancy-associated glycoproteins from water buffalo (Bubalus bubalis) placenta by Vicia villosa affinity chromatography

The present study describes the isolation and characterization of new pregnancy-associated glycoprotein molecules (PAG) from midpregnancy and late-pregnancy placentas in the water buffalo (Bubalus bubalis). After extraction, the homogenates are subjected to acid and ammonium sulfate precipitations followed by DEAE chromatography. Subsequently, the water buffalo PAG (wbPAG) from these solutions are enriched by Vicia villosa agarose (VVA) affinity chromatography. As determined by western blotting with anti-PAG sera, the apparent molecular masses of the immunoreactive bands from the VVA peaks range from 59.5 to 75.8 kDa and from 57.8 to 73.3 kDa in the midpregnancy and late-pregnancy placentas, respectively. Amino-terminal microsequencing of the immunoreactive proteins has allowed the identification of three distinct wbPAG sequences, which have been deposited in the SwissProt database: RGSXLTIHPLRNIRDFFYVG (acc. no. P85048), RGSXLTILPLRNIID (acc. no. P85049), and RGSXLTHLPLRNI (acc. no. P85050). Their comparison to previously identified proteins has shown that two of them are new because they have not been described before. Our results confirm the suitability of VVA chromatography for the enrichment of the multiple PAG molecules expressed in buffalo placenta.     

Effects of field bean (Vicia faba L. var. minor) dietary supplementation on plasma thyroid hormones, insulin, insulin-like growth factor-1 concentrations and mohair characteristics in growing Angora goat kids

Twenty-two Angora kids were used to study the effect of a dietary supplementation with field bean (Vicia faba L. var. minor) on hair follicle activity, mohair characteristics and plasma total thyroxine (T4), triiodothyronine (T3), insulin (INS) and insulin-like growth factor-1 (IGF-1) concentrations. At birth, their mothers were divided in two groups (S: supplemented, C: control), the S group being supplemented with 300 g/head/day of whole field bean. At weaning, diet of kids from S group was supplemented with 80 g/head/day of whole field bean. At weaning, secondary hair follicle activity (S: 0.91 ± 0.01, C: 0.84 ± 0.02, p < 0.01) and staple length (S: 5.82 ± 0.13 cm, C: 5.16 ± 0.14 cm, p < 0.001) were greater in the S group. At 155 days of age, secondary fibre diameter (S: 16.54 ± 0.35 μm, C: 18.09 ± 0.31 μm, p < 0.01) was higher in the C kids. Concentrations of total plasma T4 and T3 were higher (p < 0.001) in S (120 ± 12 and 4.87 ± 0.92 ng/ml respectively) than in C kids (92 ± 8 and 2.97 ± 0.77 ng/ml respectively). At weaning, T4 values were negatively correlated (p < 0.05) with the secondary fibre diameter and the T3/T4 ratio was positively correlated (p < 0.05) with the secondary to primary hair follicle ratio. At 155 days of age, both T3 and T3/T4 values were positively correlated (p < 0.01) with staple length. Plasma INS (0.11 ± 0.03 ng/ml) and total IGF-1 concentrations (232 ± 13 ng/ml) were not affected by diet. Improvements of diet quality in kids positively affected fibre diameter and length and follicle activity rate. We suggest that increased circulating thyroid hormones concentrations, but not those of INS and IGF-1, may contribute to the beneficial actions of nutritional supplementation on anatomical growth and mohair production of kids.     

Photoperiod and Soil Munition Constituent Effects on Phytoaccumulation and Rhizosphere Interactions in Boreal Vegetation

Permafrost thaw is expected to alter biogeochemistry and hydrology, potentially increasing the mobility of soil constituents. Northern latitude boreal forests where permafrost thaw is occurring also experience extreme changes in day length during the growing season. As the effects of photoperiod on plant uptake of soil constituents or interactions with the rhizosphere are unknown, our objective was to determine these interactions with three plant species from different functional groups. A tree, forb, and grass common to military training ranges in this region were grown in soil spiked with or without lead, antimony, or 2,4-dinitrotoluene and grown under 16, 20, or 24 h of light. Plant biomass, soil constituent uptake, and rhizosphere bacterial communities were compared between treatments. Photoperiod had no effect on plant uptake of any soil constituent or on rhizosphere community, indicating that plants and their associated microbial communities adapted to this environment are resilient to extremes in photoperiod. Lead uptake was not significant in any plant species and had no effect on the rhizosphere. Antimony increased the percentage composition of Saprospirales in the rhizospheres of two of the three plants, indicating an interaction between this bacterial order and antimony. Antimony uptake by white spruce (Picea glauca) was considerable, with a mean concentration of 1731 mg kg^?1 in roots, while mean shoot concentration was only 155 mg kg^?1, indicating its potential to phytostabilize this heavy metal. Although antimony had the strongest impact on the rhizosphere bacterial community, it was also readily accumulated by the grass and tree.     

Assessment of pregnancy‐associated glycoprotein (PAG) concentrations in swamp buffalo samples from fetal and maternal origins by using interspecies antisera

Pregnancy‐associated glycoproteins (PAG) constitute a large family of glycoproteins found in the outer placental epithelial cell layer of the placenta in Eutherian species. In ruminants, they are noted to be structurally closely related among the different species. This study was designed to determine PAG concentrations in maternal and fetal plasma, allantoic and amniotic fluids in buffalo species. Antisera (AS) generated in rabbits against distinct PAG molecules were used in three radioimmunoassay (RIA)‐PAG systems: RIA‐1 (antiserum raised against bovine PAG67kDa; AS#497), RIA‐2 (antiserum raised against caprine PAG55 + 62 kDa; AS#706) or RIA‐3 (antiserum raised against buffalo PAG; AS#859). Samples were collected at a slaughterhouse (n = 67). PAG concentrations determined by RIA‐2 gave significantly higher results in both allantoic and amniotic fluids (12.7 ± 2.1 ng/mL and 24.0 ± 7.3 ng/mL, respectively). Regarding maternal and fetal plasma, PAG concentrations obtained by RIA‐2 (21.8 ± 2.4 ng/mL and 20.2 ± 2.5 ng/mL, respectively) and RIA‐3 (25.0 ± 2.2 ng/mL and 21.9 ± 3.2 ng/mL, respectively) were higher than those obtained by RIA‐1 (15.5 ± 1.4 ng/mL and 16.1 ± 1.8 ng/mL, respectively). The correlation among the three systems was very high. The study clearly reveals the ability of different PAG‐RIA systems to measure PAG concentration in swamp buffalo samples.     

Effects of genotype and cryopreservation of avian semen on fertility and number of perivitelline spermatozoa

1. The fertility of freshly diluted and cryopreserved samples of semen obtained from a population of chickens selected for duration of fertility of cryopreserved spermatozoa (FS line) and its unselected control (FC line) were compared over a range of spermatozoa concentrations (10, 40, 80, and 160 × 10⁶ sperm/50 μ1 insemination).

2. The spermatozoa of the FS line had greater fertility than spermatozoa of the FC line, whether freshly diluted or cryopreserved. Cryopreservation resulted in a reduction in fertility, regardless of line. There were no significant line by genotype interactions.

3. There were fewer spermatozoa from the FG line than the FS line found in the perivitelline membrane (perivitelline spermatozoa). The increase in number of perivitelline spermatozoa with increasing sperm concentration was greater in the FS than FC line. However, the slope of the increase in sperm number in the perivitelline membrane with increasing concentrations of cryopreserved spermatozoa was zero.

4. A minimum of 10³ perivitelline spermatozoa must be found on day 2 post‐insemination for duration of fertility to exceed three days. The ability to produce spermatozoa capable of reaching the forming perivitelline membrane appears to be a quantitative, rather than a qualitative, trait and may be subject to genetic manipulation.     

Identification of pregnancy-associated glycoproteins and alpha-fetoprotein in fallow deer (Dama dama) placenta

Background

This paper describes the isolation and characterization of pregnancy-associated glycoproteins (PAG) from fetal cotyledonary tissue (FCT) and maternal caruncular tissue (MCT) collected from fallow deer (Dama dama) pregnant females. Proteins issued from FCT and MCT were submitted to affinity chromatographies by using Vicia villosa agarose (VVA) or anti-bovine PAG-2 (R#438) coupled to Sepharose 4B gel. Finally, they were characterized by SDS-PAGE and N-terminal microsequencing.

Results

Four distinct fallow deer PAG (fdPAG) sequences were identified and submitted to Swiss-Prot database. Comparison of fdPAG with PAG sequences identified in other ruminant species exhibited 64 to 83% identity. Additionally, alpha-fetoprotein was identified in fetal and maternal tissues.

Conclusion

Our results demonstrate the efficacy of VVA and bovine PAG-2 affinity chromatographies for the isolation of PAG molecules expressed in deer placenta. This is the first report giving four specific amino acid sequences of PAG isolated from feto-maternal junction (FCT and MCT) in the Cervidae family.     

A ω-secalin contained decamer shows a celiac disease prevention activity

Celiac disease (CD) is an autoimmune permanent enteropathy that is triggered in susceptible individuals after the ingestion of gluten, a storage protein fraction presents in wheat, rye and barley endosperm. Specific gluten peptides can bind to HLA-DQ2/8 and induce lamina propria CD4⁺ T cell responses causing damage of the small intestine mucosa. Recent studies suggested that beside immunodominant and toxic epitopes, wheat gluten also contains epitopes capable of preventing the mucosal response in vitro. Among them, a decapeptide (QQPQDAVQPF) from wheat was reported to have an antagonist effect on the agglutination of K562(S) cells and celiac T-cell activation, although the corresponding nucleotidic sequence remained unknown. This study was therefore designed to clone the sequence encoding the protein carrying the decapetide with CD protective properties. A ω-secalin gene encoding containing the decapeptide QQPQRPQQPF was isolated. Although the decapeptide was not identical to the one previously described, QQPQRPQQPF showed the same capability to prevent K562(S) cell agglutination and celiac mucosa immune activation induced by toxic gliadins. The ω-secalin gene was found in wheat carrying the wheat–rye chromosomal translocations 1BL.1RS. The identification of this immunomodulatory gliadin sequence, naturally occurring in cultivars of wheat toxic for celiac patients, might offer new therapeutic strategies for CD.     

Pasta made from durum wheat semolina fermented with selected lactobacilli as a tool for a potential decrease of the gluten intolerance

A pool of selected lactic acid bacteria was used to ferment durum wheat semolina under liquid conditions. After fermentation, the dough was freeze-dried, mixed with buckwheat flour at a ratio of 3:7, and used to produce the "fusilli" type Italian pasta. Pasta without prefermentation was used as the control. Ingredients and pastas were characterized for compositional analysis. As shown by two-dimensional electrophoresis, 92 of the 130 durum wheat gliadin spots were hydrolyzed almost totally during fermentation by lactic acid bacteria. Mass spectrometry matrix-assisted laser desorption/ionization time-of-flight and reversed phase high-performance liquid chromatography analyses confirmed the hydrolysis of gliadins. As shown by immunological analysis by R5-Western blot, the concentration of gluten decreased from 6280 ppm in the control pasta to 1045 ppm in the pasta fermented with lactic acid bacteria. Gliadins were extracted from fermented and nonfermented durum wheat dough semolina and used to produce a peptic-tryptic (PT) digest for in vitro agglutination tests on cells of human origin. The whole PT digests did not cause agglutination. Affinity chromatography on Sepharose-6-B mannan column separated the PT digests in three fractions. Fraction C showed agglutination activity. The minimal agglutinating activity of fraction C from the PT digest of fermented durum wheat semolina was ca. 80 times higher than that of durum wheat semolina. Pasta was subjected to sensory analysis: The scores for stickiness and firmness were slightly lower than those found for the pasta control. Odor and flavor did not differ between the two types of pasta. These results showed that a pasta biotechnology that uses a prefermentation of durum wheat semolina by selected lactic acid bacteria and tolerated buckwheat flour could be considered as a novel tool to potentially decrease gluten intolerance and the risk of gluten contamination in gluten-free products.