Gonadotropin-inhibitory hormone (GnIH) receptor gene is expressed in the chicken ovary: potential role of GnIH in follicular maturation

Gonadotropin-inhibitory hormone (GnIH), an RFamide peptide, has been found to inhibit pituitary LH secretion in avian and mammalian species. The gene encoding a putative receptor for GnIH (GnIHR) was recently identified in the chicken and Japanese quail brain and pituitary gland. GnIHR appears to be a seven-transmembrane protein belonging to a family of G-protein-coupled receptors. In the present study, we have characterized the expression of GnIHR mRNA in the chicken ovary and demonstrate that GnIHR may exert an inhibitory effect on ovarian follicular development. By RT-PCR, we detected GnIHR mRNA in the chicken testis and in the ovary, specifically both thecal and granulosa cell layers. Real-time quantitative PCR analysis revealed greater GnIHR mRNA quantity in theca cells of prehierarchial follicles compared with that of preovulatory follicles. GnIHR mRNA quantity was significantly decreased in sexually mature chicken ovaries versus ovaries of sexually immature chickens. Estradiol (E(2)) and/or progesterone (P(4)) treatment of sexually immature chickens significantly decreased ovarian GnIHR mRNA abundance. Treatment of prehierarchial follicular granulosa cells in vitro with chicken GnIH peptide significantly decreased basal but not FSH-stimulated cellular viability. Collectively, our results indicate that the ovarian GnIHR is likely to be involved in ovarian follicular development. A decrease in ovarian GnIHR mRNA abundance due to sexual maturation or by E(2) and/or P(4) treatment would implicate an inhibitory role for GnIHR in ovarian follicular development. Furthermore, GnIH may affect follicular maturation by decreasing the viability of prehierarchial follicular granulosa cells through binding to GnIHR.     

Spectroscopic evidence for Fe(II)-Fe(III) electron transfer at the iron oxide-water interface

Using the isotope specificity of 57Fe M?ssbauer spectroscopy, we report spectroscopic observations of Fe(II) reacted with oxide surfaces under conditions typical of natural environments (i.e., wet, anoxic, circumneutral pH, and about 1% Fe(II)). M?ssbauer spectra of Fe(II) adsorbed to rutile (TiO2) and aluminum oxide (Al2O3) show only Fe(II) species, whereas spectra of Fe(II) reacted with goethite (alpha-FeOOH), hematite (alpha-Fe2O3), and ferrihydrite (Fe5HO8) demonstrate electron transfer between the adsorbed Fe(II) and the underlying iron(III) oxide. Electron-transfer induces growth of an Fe(III) layer on the oxide surface that is similar to the bulk oxide. The resulting oxide is capable of reducing nitrobenzene (as expected based on previous studies), but interestingly, the oxide is only reactive when aqueous Fe(II) is present. This finding suggests a novel pathway for the biogeochemical cycling of Fe and also raises important questions regarding the mechanism of contaminant reduction by Fe(II) in the presence of oxide surfaces.     

Spectroscopic evidence for Ni(II) surface speciation at the iron oxyhydroxides-water interface

Understanding in situ metal surface speciation on mineral surfaces is critical to predicting the natural attenuation of metals in the subsurface environment. In this study, we have demonstrated the novel Ni K-edge X-ray absorption spectroscopy (XAS) measurements needed to understand Ni(ll) surface speciation in three synthetic iron oxyhydroxides (ferrihydrite, goethite, and hematite). The adsorption of Ni gradually increases with increasing pH from 5 to 8, and the adsorption edge appears at near the point of zero salt effect (PZSE) of the solids. The results of XAS analysis indicate four different Ni inner-sphere surface species are present. While total Ni surface species in hematite at pH 6.85 surfaces consist of approximately 63% face-sharing (interatomic distance of Ni-Fe (R(Ni-Fe)) approximately 2.9 A) and approximately 37% corner-sharing (R(Ni-Fe) approximately 4.0 A) surface species on iron octahedra, a combination of two different edge-sharing (between NiO6 and FeO6 octahedra, in chains or in rows) and corner-sharing surface species are observed in goethite and ferrihydrite at pH 5.09-6.89. In ferrihydrite, approximately 70% of surface species are edge-sharing surface species (in chains) (R(Ni-Fe) approximately 3.0 A), followed by approximately 30% of edge-sharing species (in rows) (R(Ni-Fe) approximately 3.2 A) and approximately 3-5% of corner-sharing surface species (R(Ni-Fe) approximately 4.0 A). Goethite contains approximately 54% edge-sharing (R(Ni-Fe) approximately 3.0 A), approximately 26% edge-sharing (R(Ni-Fe) approximately 3.2 A), and 20% corner-sharing surface species. These findings indicate that the reactivity and surface speciation of Ni are sensitive to the crystallinity of iron oxyhydroxides. The spectroscopic evidence for multi-Ni surface speciation should be factored into predictions of the transport of Ni in soil-water environments.     

Quantification of healthy follicles in the neonatal and adult mouse ovary: evidence for maintenance of primordial follicle supply

Proliferation and partial meiotic maturation of germ cells in fetal ovaries is believed to establish a finite, non-renewable pool of primordial follicles at birth. The supply of primordial follicles in postnatal life should be depleted during folliculogenesis, either undergoing atresia or surviving to ovulation. Recent studies of mouse ovaries propose that intra- and extraovarian germline stem cells replenish oocytes and form new primordial follicles. We quantified all healthy follicles in C57BL/6 mouse ovaries from day 1 to 200 using unbiased stereological methods, immunolabelling of oocyte meiosis (germ cell nuclear antigen (GCNA)) and ovarian cell proliferation (proliferating cell nuclear antigen (PCNA)) and electronmicroscopy. Day 1 ovaries contained 7924+/-1564 (s.e.m.) oocytes or primordial follicles, declining on day 7 to 1987+/-203, with 200-800 oocytes ejected from individual ovaries on that day and day 12. Discarded oocytes and those subjacent to the surface epithelium were GCNA-positive indicating their incomplete meiotic maturation. From day 7 to 100 mean numbers of primordial follicles per ovary were not significantly depleted but declined at 200 days to 254+/-71. Mean numbers of all healthy follicles per ovary were not significantly different from day 7 to 100 (range 2332+/-349-3007+/-322). Primordial follicle oocytes were PCNA-negative. Occasional unidentified cells were PCNA-positive with mitotic figures observed in the cortex of day 1 and 12 ovaries. Although we found no evidence for ovarian germline stem cells, our data support the hypothesis of postnatal follicle renewal in postnatal and adult ovaries of C57BL/6 mice.     

The roles of risk and ambiguity in the adoption of the system of rice intensification (SRI): evidence from Indonesia

Given the recognized yield-enhancing potential of the System of Rice Intensification (SRI), its low adoption and high discontinuance rates in some locales are puzzling. Combining experimental measures of risk and ambiguity aversion with household-level and plot-level panel data collected in rural Indonesia, this study empirically explores factors shaping SRI’s adoption and discontinuance. Employing multivariate and Heckman probit models to control unobserved heterogeneities, we find that farmers’ risk aversion significantly reduces their likelihood of using all individual SRI practices. However, once the effects of risk aversion on the use of SRI in the previous year are statistically controlled, risk aversion does not significantly explain farmers’ subsequent decisions to continue or discontinue SRI practices. Farmers’ ambiguity preferences play no significant role in decisions to use most practices, except alternate wetting and drying, which requires proper coordination of irrigation among neighboring farmers and thus amplifies the uncertainty of effective implementation. The results also show that access to irrigation is a significant factor in the use of SRI and its continuance. Moreover, as SRI requires greater input of labor and therefore curtails time for alternative household activities, including off-farm work, family composition is a significant factor determining its adoption and continuing use. Although these findings are not necessarily generalizable, our study expands the existing knowledge of factors underlying SRI’s slow diffusion.     

Antioxidant activity of phenolic compounds: from in vitro results to in vivo evidence

Over last decade an increasing interest for antioxidants in foods has arisen. The healthy properties of antioxidants related to the prevention of degenerative diseases are the main cause of this boom. An antioxidant prevents the oxidation process, the initial step of development of degenerative diseases, cancer and many others. Literature encompasses analytical methodology development to assess antioxidant properties of foods and beverages. The screening of antioxidant activity of foodstuffs is the subject of a large number of articles. Special interest has been addressed to wine, tea and chocolate. However, the crucial key in the prevention of disease is the action these antioxidants exert after their consumption. Studies involving human subjects are scarce due to the requirements of availability of volunteers and conditions to test are limited. This review summarizes data related to in vitro antioxidant activity of foods, emphasizing the main role of phenolic compounds. A critical comparison is realized between the biological significance of these values and the biological significance of in vivo measurements. In addition, the Plasma Antioxidant Capacity is evaluated and selected as biomarker for in vivo antioxidant status of human organism. In a second part, data collected from different intervention studies performed up to date are compiled and discussed. This review summarized data related to in vitro antioxidant activity of foods, emphasizing the main role of phenolic compounds. A critical comparison is realized between the biological significance of these values and the biological significance of in vivo measurements. In addition, the Plasma Antioxidant Capacity is evaluated and selected as biomarker for in vivo antioxidant status of human organism. In a second part, data collected from different intervention studies performed up to date are compiled and discussed. The original contribution of this work is to compile data of Plasma Antioxidant Capacity after dietetic intervention studies taking into account the portion of food ingested. In addition, we calculated the antioxidant compounds content (phenolic content, ascorbic acid, vitamin E and carotenoids) contained in each food ingested to evaluate better their impact in Plasma Antioxidant Capacity. Intervention studies are grouped by the length of intervention and type of food ingested. Results reported in literature reveal that the increment in Plasma Antioxidant Capacity largely depends on analytical method used.     

Photocatalytic oxidation of arsenic(III): evidence of hydroxyl radicals

Arsenic contamination has been found in the groundwater of several countries. Photocatalysis can rapidly oxidize arsenite (As(III)) to less labile and less toxic arsenate (As(V)), which then can be removed by adsorption onto photocatalyst surfaces. This study investigates the photocatalytic oxidation of As(III) to As(V) as a function of As(III) concentration, pH, catalyst loading, light intensity, dissolved oxygen concentration, type of TiO2 surfaces, and ferric ions to understand the kinetics and the mechanism of As(III) oxidation in the UV/TiO2 system. Photocatalytic oxidation of As(III) to As(V) takes place in minutes and follows zero-order kinetics. Benzoic acid (BA) was used as a hydroxyl radical (.OH) scavenger to provide evidence for the .OH as the main oxidant for oxidation of As(III). The .OH radical was independently generated by nitrate photolysis, and kinetics of As(III) oxidation by the .OH radical was determined. Formation of salicylic acid (SA) from the oxidation of BA by .OH also demonstrates the involvement of .OH in the mechanism of As(III) oxidation. The effect of Fe(III) on As(III) oxidation at different pH values with and without TiO2 under UV light was examined. The results suggest that .OH is the dominant oxidant for As(III) oxidation. Two commercially available TiO2 suspensions, Degussa P25 and Hombikat UV100, were tested for the removal of arsenic through oxidation of As(III) to As(V) followed by adsorption of As(V) onto TiO2 surfaces. Results showed that complete removal of arsenic below the World Health Organization drinking water limit of 10 microg/L could be achieved.     

X-ray absorption spectroscopic evidence for the formation of Pb(II) inner-sphere adsorption complexes and precipitates at the calcite-water interface

Combined batch sorption and in situ X-ray absorption spectroscopy provide direct assessment of the mechanisms for Pb(II) sorption atthe calcite--water interface under low-temperature conditions. At low metal concentration, 1 microM initial Pb, X-ray absorption fine structure data indicate the formation of Pb mononuclear inner-sphere complexes at the surface. A first-shell Pb-O bond length of 2.34 A is consistent with nearest oxygen neighbors in 3- or 4-fold coordination with a distorted trigonal pyramidal or square pyramidal geometry with a stereochemically active electron lone pair. For high initial Pb concentrations, 20 and 60 microM Pb, precipitation of hydrocerussite and cerussite secondary phases dominates Pb partitioning. At 5 and 10 microM initial Pb, the sorption mechanism is dual in nature with persistence of the mononuclear adsorption complex combined with precipitation of a cerussite phase occurring prior to saturation of theoretically available surface sites. The formation of inner-sphere complexes implies strong metal interactions with the surface-the mechanistic reason for the affinity of Pb for calcite as observed in macroscopic studies. The geometry of the adsorbed complex can influence Pb coprecipitation, as a change to octahedral coordination is required for incorporation into calcite. The results provide the basis for predictions of Pb sequestration by calcite in natural systems.     

Expression of the long (OB-RB) and short (OB-RA) forms of the leptin receptor throughout the oestrous cycle in the mature rat ovary

Leptin is secreted by adipocytes and exerts its effects by interacting with the long form of the leptin receptor, OB-RB. The leptin protein and leptin receptors have been localized in the ovary, and acute leptin treatment directly inhibits ovulation in the rat ovary. It was hypothesized that expression of the leptin receptor gene varies throughout the oestrous cycle to modulate the sensitivity of the ovary to leptin. In this study, expression of genes for the long and short isoforms of the leptin receptor in the adult ovary was investigated at different stages of the rat oestrous cycle. Vaginal cytology was used to determine the stage of the oestrous cycle. Ovaries were collected and RNA was extracted for real-time RT-PCR analysis of leptin receptor gene expression. OB-RB gene expression was low in pro-oestrus (3.13 +/- 0.18 fg RNA per microg total DNA) and dioestrus II (2.52 +/- 0.19 fg RNA per microg total DNA) of the oestrous cycle, whereas expression was high in oestrus (5.9 +/- 0.27 fg RNA per microg total DNA) and dioestrus I (4.6 +/- 0.24 fg RNA per microg total DNA) (P < 0.001). Expression of the gene for the short form of the leptin receptor (OB-RA) was at a maximum in dioestrus I (65.5 +/- 0.8 fg RNA per ng total DNA), high in oestrus (39.0 +/- 0.8 fg RNA per ng total DNA) and low at pro-oestrus (5.0 +/- 0.2 fg RNA per ng total DNA) and dioestrus II (1.1 +/- 0.09 fg RNA per ng total DNA) (P < 0.001). Plasma oestradiol concentrations (pg ml-1) were highest at pro-oestrus (19.38 +/- 1.3), and similar at the remaining three stages studied (oestrus: 13.7 +/- 1.9; dioestrus I: 12.4 +/- 1.0; dioestrus II: 10.3 +/- 0.9) (P < 0.05). Plasma progesterone concentrations (ng ml-1) were higher in the luteal phases of the oestrous cycle (dioestrus I: 18.6 +/- 2.3; dioestrus II: 14.7 +/- 2.5) than during pro-oestrus (5.12 +/- 0.6) and oestrus (5.9 +/- 0.8) (P < 0.05). Plasma leptin concentrations were detectable only in pro-oestrus (0.35 +/- 0.05 ng ml(-1)) and were below the detection limit of the assay at other stages of the oestrous cycle. In summary, mRNA content for the long and short isoforms of the leptin receptor is lower in pro-oestrus and dioestrus II than in oestrus and dioestrus I of the rat oestrous cycle. The fluctuations in leptin receptor mRNA content may be a response to the concentrations of circulating steroid hormones and leptin. This research supports the initial hypothesis and shows that ovarian leptin receptor concentrations vary throughout the oestrous cycle in response to the changing environment of the ovary.     

In vivo evidence for non-universal usage of the codon CUG in Candida maltosa

An alkane-assimilating yeast Candida maltosa had been studied in order to establish systems suitable for biotransformation of hydrophobic compounds. However, functional expression of heterologous genes tested for this purpose had not been successful in several cases. On the other hand, it had been reported that the codon CUG, a universal leucine codon, is read as serine in C. cylindracea. The same altered codon usage had also been suggested by in vitro experiments in some Candida yeasts which are phylogenetically closely related to C. maltosa. In this study we have shown that the failure in functional expression of a heterologous gene is due to the fact that the codon CUG is read as serine in C. maltosa. This conclusion was drawn from the following experimental results: (1) when a cytochrome P450 gene of C. maltosa containing a CTG codon was expressed in C. maltosa, the corresponding amino acid was found to be serine, and not leucine; (2) a tRNA gene with an almost identical structure to that of the tRNA ^SerCAG gene of C. albicans could be isolated from the genome of C. maltosa; (3) the Saccharomyces cerevisiae URA3 gene, which has one CTG codon, could not complement the ura3 mutation of C. maltosa as itself, but when the CTG codon was changed to another leucine codon, CTC, the mutated gene could complement the ura3 mutation. The last result is the first example of succeeding in functional expression of a heterologous gene in Candida species having an altered codon usage by changing the CTG codon in the gene to another codon. The nucleotide sequence datum reported in this paper will appear in the GSDB, DDBJ, EMBL and NCBI nucleotide sequence databases with the Accession Number D26074.     

In vitro and in vivo evidence for a satiating effect of fish protein hydrolysate obtained from blue whiting (Micromesistius poutassou) muscle

To find appetite suppressive molecules derived from fish protein hydrolysates, both in vitro and in vivo experiments were performed in order to demonstrate that hydrolysates produced from blue whiting muscle (BWMH) possess satiating properties. Here we demonstrated for the first time that a protein hydrolysate obtained from marine source was able to enhance cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1) secretion in STC-1 cell line. To demonstrate that these in vitro activities also exist in vivo, we investigated the effect of BWMH preload administration in rats and its repercussion on food intake and metabolic plasma marker levels. Results showed that BWMH reduced the short term food intake which was correlated to an increase in the CCK and GLP-1 plasma levels. Moreover it was demonstrated that the chronic administration of BWMH led to a decrease in the body weight gain.     

益生菌中胆盐水解酶(BSHs)研究进展

探讨益生菌及其制品中胆盐水解酶降血压的理论依据，并阐述了益生菌胆盐水解酶的特性，对菌体自身的影响和其对宿主发挥的功能特性，展望了胆盐水解酶的未来吁究方向和发展趋势。     

Expression of alternative oxidase gene (aox1) at the stage of single-cell conidium in citric acid-producing Aspergillus niger

Mycelia of citric acid-producing Aspergillus niger WU-2223L show cyanide-insensitive respiration catalyzed by alternative oxidase. In this study, the constitutive expression of alternative oxidase gene (aox1) even at the stage of single-cell conidium in A. niger WU-2223L was found using the visual expression analysis system of aox1 with green fluorescent protein under microscopy observation.     

The in vivo antioxidant and antifibrotic properties of green tea (Camellia sinensis,Theaceae)

The in vivo antioxidant and antifibrotic properties of green tea (Camellia sinensis, Theaceae) were investigated with a study of carbon tetrachloride (CCl₄)-induced oxidative stress and hepatic fibrosis in male ICR mice. Oral administration of green tea extract at doses of 125, 625 and 1250 mg/kg for 8 weeks significantly reduced (p < 0.05) the levels of thiobarbituric acid-reactive substances (TBARS) and protein carbonyls in the liver by at least 28% compared with that was induced by CCl₄ (1 mL/kg) in mice. Moreover, green tea extract administration significantly increased (p < 0.05) the activities of catalase, glutathione peroxidase (GSH-Px) and glutathione reductase (GSH-Rd) in the liver. Our study found that oral administration of green tea extract prevented CCl₄-induced hepatic fibrosis, as evidenced by a decreased hydroxyproline level in the liver and a reduced incidence of hepatic fibrosis by histological observations. These results indicate that green tea exhibits potent protective effects against CCl₄-induced oxidative stress and hepatic fibrosis in mice by inhibiting oxidative damage and increasing antioxidant enzyme activities.     

Identification of a single nucleotide polymorphism at intron 16 of the caprine acyl-coenzyme A: diacylglycerol acyltransferase 1 (DGAT1) gene

The DGAT1 gene encodes a microsomal enzyme that catalyses the only committed step in triacylglycerol synthesis by joining diacylglycerol and fatty acyl coenzyme A. In cattle, a K232A substitution in the DGAT1 molecule has a significant effect on enzyme activity and milk fat content. The prominent role of this gene in lipid metabolism led us to undertake the structural characterization of DGAT1 in goats. In this way, we have sequenced a 1552 bp fragment of the goat DGAT1 cDNA, which encompasses most of the coding sequence (from exon 1 to 17), and a genomic fragment covering exons 12 to 17. Multiple alignment of the goat DGAT1 sequences revealed the existence of a single nucleotide polymorphism (SNP) involving a T to C substitution at intron 16. We optimized a primer extension based genotyping method that allowed us to determine that the C variant is a minority allele with frequencies ranging from 0.062 (Murciano-Granadina) to 0.109 (Malague?a). This SNP, although not expected to have any functional effect, might be useful as a genetic marker in association studies to detect additional DGAT1 polymorphisms which might influence fat milk content and other traits of economic interest.