Identification of extracellular vesicles and characterization of miRNA expression profiles in human blastocoel fluid.

In this study, for the first time, we demonstrated the presence of microRNAs and extracellular vesicles in human blastocoel fluid. The bioinformatic and comparative analyses identified the biological function of blastocoel fluid microRNAs and suggested a potential role inside the human blastocyst. We found 89 microRNAs, expressed at different levels, able to regulate critical signaling pathways controlling embryo development, such as pluripotency, cell reprogramming, epigenetic modifications, intercellular communication, cell adhesion and cell fate. Blastocoel fluid microRNAs reflect the miRNome of embryonic cells and their presence, associated with the discovery of extracellular vesicles, inside blastocoel fluid, strongly suggests their important role in mediating cell communication among blastocyst cells. Their characterization is important to better understand the earliest stages of embryogenesis and the complex circuits regulating pluripotency. Moreover, blastocoel fluid microRNA profiles could be influenced by blastocyst quality, therefore, microRNAs might be used to assess embryo potential in IVF cycles.

Expression analysis of TFIID in single human oocytes: new potential molecular markers of oocyte quality.

Molecular characterization of human female gametes should make it easier to understand the basis of certain infertility disorders. Biologically significant mRNAs have been analysed in single oocytes to search for molecular biomarkers of oocyte quality. Initial analysis was focused on mRNA for proteins involved in cell growth and cycle control, specifically those encoding members of the general transcription apparatus such as the subunits of the general transcription factor TFIID. This heteromultimeric protein, comprising about 15 subunits, is the most important general transcription factor of the second class. These proteins are essential for the initiation of transcription of protein-coding genes, so they must be present in mature oocytes for mRNA synthesis during the first phases of embryonic development. Semi-quantitative reverse transcriptionpolymerase chain reaction was used to identify different TFIID subunits in single oocytes and to search for differences in expression as compared with control tissues. The data show that the mRNAs for most TFIID subunits are indeed synthesized in oocytes, but their expression profiles differ markedly. TATAbox-binding protein associated factor 4B (TAF4B), TAF5 and TATAbox-binding protein-like 2 (TBPL2) are expressed at higher levels in oocytes than in control tissues. It is suggested that they could be used as biomarkers of oocyte quality.

Intramuscular fatty acid composition of lambs given a tanniniferous diet with or without polyethylene glycol supplementation.

The aim of this trial was to investigate the effects that dietary tannins have on lamb intramuscular fatty acids. Twenty-seven lambs were divided into three homogeneous groups: control group, receiving commercial concentrate based on maize; tannin group, fed a diet based on carob pulp (45% as fed basis); PEG group, receiving the same diet as the latter with addition of 42g/kg of polyethylene glycol (PEG, a binding agent that eliminates the effects of condensed tannins). The duration of the trial was 45 d. Intramuscular fatty acids were measured in the longissimus dorsi muscle. The isomer cis-9 trans-11 of linoleic acid (conjugated linoleic acid or CLA) and linolenic acid were higher in the longissimus muscle fat from animals fed the control diet compared to the other groups (P<0.0005); these fatty acids were higher in the fat from animals fed the carob diet supplemented with PEG compared to those fed the same diet without PEG (P<0.05). trans-Vaccenic acid (C18:1 trans-11) was found at higher concentration in fat from control and PEG lambs compared to tannin lambs (P<0.01); the CLA/C18:1 trans-11 ratio was lower in lambs fed control and PEG diets than in tannin-fed animals (P<0.05).

Lamb meat quality as affected by a natural or artificial milk feeding regime.

Fourteen Barbaresca lambs were divided into two groups of seven at 24h from birth and reared exclusively on natural milk (NR) or on artificial milk (AR). Lambs were slaughtered at 40 days of age. The NR group tended (P<0.10) to grow faster, thus resulting in a higher (P<0.10) carcass weight as compared to the AR group. AR meat was darker (P<0.05), leaner (P<0.001) and with a higher moisture (P<0.10) and ash (P<0.05) content than its counterpart. The proportion of polyunsaturated fatty acids was higher (P<0.01), while that of saturated lower (P<0.01) in meat from the AR group, resulting in a higher (P<0.01) polyunsaturated/saturated ratio. Linoleic acid content (P<0.001) and its n-6 series derivatives, except 9-12 trans C18:2 n-6 (P<0.001), increased more in the AR group meat than in the NR group. On the other hand, α-linolenic (P<0.001) and other n-3 series fatty acids were higher in meat from the NR group than in the AR group, leading to a lower (P<0.001) n-6/n-3 ratio. Furthermore, 9 cis 11 trans CLA was higher (P<0.001) in NR meat compared to AR meat. Finally, a milk-feeding regime exclusively based on artificial milk adversely affected the dietetic value of lamb meat compared to a natural rearing system, reducing the level of desirable fatty acids such as n-3 series and CLA.

Citrus pulp as an ingredient in ostrich diet: effects on meat quality.

The purpose of this trial was to study the effects of citrus pulp inclusion in ostrich diets on meat quality, evaluated on iliofibularis and gastrocnemius muscles. M. iliofibularis had a lower ultimate pH (P<0.05) and was lighter in colour (P<0.05) than M. gastrocnemius. The latter had higher moisture (P<0.05) and lower crude protein contents (P<0.05) as compared to M. iliofibularis. M. gastrocnemius had a lower proportion of C14:0 (P<0.05), C16:0 (P<0.05) and C16:1 (P<0.05) and a higher percentage of C18:0 (P<0.05), C20:2ω6 (P<0.05), C20:4ω6 (P<0.05) and C20:5ω3 (P<0.05) than M. iliofibularis. Citrus pulp diet increased (P<0.05) meat ultimate pH and reduced cooking losses (P<0.05) as compared to the control diet. Meat from animals given citrus pulp had lower crude fat (P<0.05) and ash percentages (P<0.05) in comparison to that from the control group. The proportions of intramuscular saturated and monounsaturated fatty acids were lower (P<0.05) in the citrus pulp group as compared to the control one. Meat from the citrus pulp treatment group had a higher (P<0.05) percentage of polyunsaturated fatty acids than meat from the control treatment. In particular, meat from ostriches given citrus pulp diet exhibited higher C18:2ω6 (P<0.05) and C20:4ω6 (P<0.05) contents that accounted for a higher ω6/ω3 ratio (P<0.05) as compared to the control group. Overall, the use of a citrus pulp-included diet in ostrich feeding did not adversely affect meat quality and, therefore, citrus pulp seems to be a possible ingredient to reduce feeding costs.