Effect of n-3 and n-6 fatty acid supplementation on fetal, gestation and parturition in pregnant Sprague Dawley rats

The aim of this study was to evaluate the effect of different ratio of n-6:n-3 on gestation and parturition as well as to determine the hormone concentration on plasma of the pregnant rats. At the end of the trial period, blood plasma was collected and analysed for progesterone, estradiol and prostaglandin F2α hormone (PGF2α) concentrations, and fatty acids profile. The results indicated that the group with a very low ratio of n-6:n3 fatty acids (diet 1) showed increased concentration of progesterone in the plasma (P<0.05), while the group with high ratio of n6:n3 fatty acids (diet 3) with high arachidonic acid (AA) showed significantly (P<0.05) increased concentration of estradiol and PGF2α in the plasma. The n-3 polyunsaturated fatty acids (PUFA) in plasma of diet 1 group were significantly (P<0.05) higher than the other treatment diet groups. The total n-6 PUFA was significantly higher (P<0.05) in diet 3 group as compared to diet 1 group. In contrast, the number of implanted embryos was significantly lower (P<0.05) in the diet 3 group at 15 days of gestation, while, the litter size were significantly lower in the diet 2 group and diet 3 group by 4.57 and 1.00 folds respectively, as compared to the control group (diet 4). In conclusion, for the rat with very low ratio on diet 1, the n-6:n-3 ratio satisfied the requirement for the growth of mother and fetuses but was inadequate for the normal process of parturition, probably through inadequate production of the prostaglandins involved.Key words: n-6:n-3, plasma fatty acids, progesterone, estradiol, prostaglandin production, pregnant rat

Effect of dietary n-6 to n-3 polyunsaturated fatty acid ratio on prostaglandin plasma levels and genes expression peroxisome proliferator-activated receptor (PPAR) in pregnant Sprague Dawley rats

The peroxisome proliferator-activated receptors (PPARs) are a family of nuclear transcription factors thought to act as receptors for polyunsaturated fatty acids and to reduce production of series 2 prostaglandins (PG). The objective of this study was to investigate the effect of different ratio n-6:n-3 on the PPAR expression of rats endometrial tissue. The findings obtained from this study showed significant induction of PPARδ mRNA levels in endomatral cells treatment 1:1 group by 1.38 fold compared with the PPARδ mRNA levels in endomatral cells treatment 30:1 group. This induction was due to the cellular demands for prostaglandin were high in the endometrial cells when cells were treated with high ratio n6:n3 on 30:1 group, thus, resulting in an increase in both prostaglandin PGE2 and PGF2α production by induction of PPARδ genes. On the other hand, treatment 1:1 group and control group of endometrial cells did not show any significant changes in mRNA level of PPARδ, compared with treatment ratio n6:n3 on 6:1 group and treatment high ratio n6:n3 on 30:1 group of the endometrial cells. These findings show that inhibition of uterine PGF2α synthesis by n-3 fatty acids may depend on the amount of n-6 fatty acids reaching the target tissue. In conclusion, PPARδ function in the response of rat endometrium to long chain n-6:n3 polyunsaturated fatty acids.Key words: Polyunsaturated fatty acid, gene expression, peroxisome proliferator-activated receptor, prostaglandin, pregnancy rat

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Analysis of the melon (Cucumis melo) small RNAome by high-throughput pyrosequencing

Abstract Background Melon (Cucumis melo L.) is a commercially important fruit crop that is cultivated worldwide. The melon research community has recently benefited from the determination of a complete draft genome sequence and the development of associated genomic tools, which have allowed us to focus on small RNAs (sRNAs). These are short, non-coding RNAs 21-24 nucleotides in length with diverse physiological roles. In plants, they regulate gene expression and heterochromatin assembly, and control protection against virus infection. Much remains to be learned about the role of sRNAs in melon. Results We constructed 10 sRNA libraries from two stages of developing ovaries, fruits and photosynthetic cotyledons infected with viruses, and carried out high-throughput pyrosequencing. We catalogued and analysed the melon sRNAs, resulting in the identification of 26 known miRNA families (many conserved with other species), the prediction of 84 melon-specific miRNA candidates, the identification of trans-acting siRNAs, and the identification of chloroplast, mitochondrion and transposon-derived sRNAs. In silico analysis revealed more than 400 potential targets for the conserved and novel miRNAs. Conclusion We have discovered and analysed a large number of conserved and melon-specific sRNAs, including miRNAs and their potential target genes. This provides insight into the composition and function of the melon small RNAome, and paves the way towards an understanding of sRNA-mediated processes that regulate melon fruit development and melon-virus interactions.This work was supported by grants AGL2009-07552/AGR, BIO2006-13107 (Ministerio de Ciencia e Innovación, Spain) and MELONOMICS (Fundación Genoma España, Spain).Peer Reviewe

Decellularized Matrix from Tumorigenic Human Mesenchymal Stem Cells Promotes Neovascularization with Galectin-1 Dependent Endothelial Interaction

BACKGROUND: Acquisition of a blood supply is fundamental for extensive tumor growth. We recently described vascular heterogeneity in tumours derived from cell clones of a human mesenchymal stem cell (hMSC) strain (hMSC-TERT20) immortalized by retroviral vector mediated human telomerase (hTERT) gene expression. Histological analysis showed that cells of the most vascularized tumorigenic clone, -BD11 had a pericyte-like alpha smooth muscle actin (ASMA+) and CD146+ positive phenotype. Upon serum withdrawal in culture, -BD11 cells formed cord-like structures mimicking capillary morphogenesis. In contrast, cells of the poorly tumorigenic clone, -BC8 did not stain for ASMA, tumours were less vascularized and serum withdrawal in culture led to cell death. By exploring the heterogeneity in hMSC-TERT20 clones we aimed to understand molecular mechanisms by which mesenchymal stem cells may promote neovascularization. METHODOLOGY/PRINCIPAL FINDINGS: Quantitative qRT-PCR analysis revealed similar mRNA levels for genes encoding the angiogenic cytokines VEGF and Angiopoietin-1 in both clones. However, clone-BD11 produced a denser extracellular matrix that supported stable ex vivo capillary morphogenesis of human endothelial cells and promoted in vivo neovascularization. Proteomic characterization of the -BD11 decellularized matrix identified 50 extracellular angiogenic proteins, including galectin-1. siRNA knock down of galectin-1 expression abrogated the ex vivo interaction between decellularized -BD11 matrix and endothelial cells. More stable shRNA knock down of galectin-1 expression did not prevent -BD11 tumorigenesis, but greatly reduced endothelial migration into -BD11 cell xenografts. CONCLUSIONS: Decellularized hMSC matrix had significant angiogenic potential with at least 50 angiogenic cell surface and extracellular proteins, implicated in attracting endothelial cells, their adhesion and activation to form tubular structures. hMSC -BD11 surface galectin-1 expression was required to bring about matrix-endothelial interactions and for xenografted hMSC -BD11 cells to optimally recruit host vasculature

Neonatal exendin-4 reduces growth, fat deposition and glucose tolerance during treatment in the intrauterine growth-restricted lamb

BACKGROUND IUGR increases the risk of type 2 diabetes mellitus (T2DM) in later life, due to reduced insulin sensitivity and impaired adaptation of insulin secretion. In IUGR rats, development of T2DM can be prevented by neonatal administration of the GLP-1 analogue exendin-4. We therefore investigated effects of neonatal exendin-4 administration on insulin action and β-cell mass and function in the IUGR neonate in the sheep, a species with a more developed pancreas at birth. METHODS Twin IUGR lambs were injected s.c. daily with vehicle (IUGR+Veh, n = 8) or exendin-4 (1 nmol.kg-1, IUGR+Ex-4, n = 8), and singleton control lambs were injected with vehicle (CON, n = 7), from d 1 to 16 of age. Glucose-stimulated insulin secretion and insulin sensitivity were measured in vivo during treatment (d 12–14). Body composition, β-cell mass and in vitro insulin secretion of isolated pancreatic islets were measured at d 16. PRINCIPLE FINDINGS IUGR+Veh did not alter in vivo insulin secretion or insulin sensitivity or β-cell mass, but increased glucose-stimulated insulin secretion in vitro. Exendin-4 treatment of the IUGR lamb impaired glucose tolerance in vivo, reflecting reduced insulin sensitivity, and normalised glucose-stimulated insulin secretion in vitro. Exendin-4 also reduced neonatal growth and visceral fat accumulation in IUGR lambs, known risk factors for later T2DM. CONCLUSIONS Neonatal exendin-4 induces changes in IUGR lambs that might improve later insulin action. Whether these effects of exendin-4 lead to improved insulin action in adult life after IUGR in the sheep, as in the PR rat, requires further investigation.Kathryn L. Gatford, Siti A. Sulaiman, Saidatul N. B. Mohammad, Miles J. De Blasio, M. Lyn Harland, Rebecca A. Simmons, Julie A. Owen