Genetic diversity and phylogeny of pearl millets [Pennisetum glaucum (L.) R. Br.] based on chloroplast trnL-F region

Local varieties of pearl millets in Saudi Arabia are heat- and drought-tolerant and are grown under various environments. Here, we evaluated the diversity of six local varieties based on sequencing and analysis of the trnL-F region of their chloroplast DNA that includes the genes of tRNA^Leu and tRNA^Phe and the intergenic spacer between them, trnL-F. The obtained sequences were 1171–1172 bp in length, including the complete sequences of the tRNA^Leu, trnL-F and tRNA^Phe regions. Three variable nucleotide sites were observed in the sequences of the Indian variety, whereas for each of the three varieties, Tihamah, Jazan, and Sayah, a single variable site was observed. The varieties, Yemeni and Baydhan, were identical for the entire analyzed sequence. The secondary structure of tRNA^Leu intron of Indian variety was distinct from the other five varieties, while Tihamah and Jazan were distinct in the secondary structure of trnL-F spacer. Additionally, Indian was distinct in the secondary structure of tRNA^Leu whereas Sayah and Jazan were distinct in the secondary structure of tRNA^Phe. Phylogenetic analysis with NCBI-retrieved accessions of the Pennisetum glaucum revealed close relationships between the investigated varieties with an exception of the Indian, which clustered separately from the other five varieties. The nucleotide polymorphism analysis revealed higher nucleotide diversity in the spacer between tRNA^Leu and tRNA^Phe than in the tRNA^Leu intron across the six local varieties. This study provides insights into the genetic diversity within local pearl millets, which is important for the conservation of the local varieties that exhibit exceptional adaptation to the harsh climate conditions in the Arabian Peninsula.

     

Viscosity of oceanic asthenosphere inferred from remote triggering of earthquakes

A sequence of large interplate earthquakes from 1952 to 1965 along the Aleutian arc and Kurile-Kamchatka trench released accumulated stresses along nearly the entire northern portion of the Pacific Plate boundary. The postseismic stress evolution across the northern Pacific and Arctic basins, calculated from a viscoelastic coupling model with an asthenospheric viscosity of 5 x 10(17) pascal seconds, is consistent with triggering of oceanic intraplate earthquakes, temporal patterns in seismicity at remote plate boundaries, and space-based geodetic measurements of anomalous velocity over an area 7000 by 7000 kilometers square during the 30-year period after the sequence.     

Osteopontin in Seminal Plasma and Sperm Membrane of Dogs

The objective of this work was to describe the presence of osteopontin (OPN) in canine seminal plasma and sperm membranes. A pool of seminal plasma and sperm membrane extract from 30 dogs was used. Polyacrylamide electrophoresis gels were performed and the bands were transferred to nitrocellulose paper and Western blot was undertaken using an antibody anti-OPN. Two and 12 bands were marked in the seminal plasma (77.2 and 15.6 kDa) and sperm membrane extracts (70.6–26.6 kDa), respectively. However, from 12 marked bands in the sperm membrane extract, only three (46.4, 37.7 and 36.5 kDa) were strongly marked. We conclude that, seminal plasma and sperm membranes from dogs contain different isoforms of OPN; yet, further studies will be necessary to determine their function in this species.     

Effects of Retinoids on the In Vitro Development of Capra Hircus Embryos to Blastocysts in Two Different Culture Systems

The objective of this study was to evaluate the effect of retinol (RT) and retinoic acid (RA) on the in vitro development of pre‐implantation goat embryos cultured in potassium simplex optimized medium or synthetic oviduct fluid or cocultured in oviductal cells monolayer either in potassium simplex optimized medium or synthetic oviduct fluid. A total of 2407 cumulus‐oocyte complexes were aspirated from 2 to 6 mm ovarian follicles from slaughtered animals. Selected cumulus‐oocyte complexes were subjected to in vitro maturation in TCM 199 for 24 h at 39°C in an atmosphere of 5% (v/v) CO₂ in humidified air. In vitro fertilization was performed in modified defined medium. Eighteen hours after in vitro fertilization, cumulus cells were removed and presumptive zygotes were randomly distributed into experimental groups. In Experiment 1, presumptive zygotes were cultured in potassium simplex optimized medium, potassium simplex optimized medium + RT, potassium simplex optimized medium + retinoic acid, synthetic oviduct fluid, synthetic oviduct fluid + RT and synthetic oviduct fluid + RA at 39°C in a humidified atmosphere of 5% (v/v) CO₂, 5% (v/v) O₂ and 90% (v/v) N₂. In Experiment 2, presumptive zygotes were cocultured in potassium simplex optimized medium + oviductal cells monolayer, potassium simplex optimized medium + RT + oviductal cells monolayer, potassium simplex optimized medium + RA + oviductal cells monolayer, synthetic oviduct fluid + oviductal cells monolayer, synthetic oviduct fluid + RT + oviductal cells monolayer and synthetic oviduct fluid + RA + oviductal cells monolayer in an atmosphere of 5% (v/v) CO₂ in humidified air. In both experiments, media were partially changed on day 2 after in vitro fertilization and unfertilized oocytes were excluded from the experiment. Embryos were cultured or cocultured for 8 days. In Experiment 1, there was no effect of RT or RA supplementation on the proportion of oocytes that reached the morula or blastocyst stages. By contrast, Experiment 2 demonstrated that the addition of 0.28 μg/ml RT and 0.5 μm RA to the embryo culture media stimulated (p < 0.05) development to the morula and blastocyst stages under the coculture conditions tested. In conclusion, retinoids play an important role in pre‐implantation development of goat embryos and can be used to enhance in vitro embryo production.     

Promotion of the cycling of diet-enhancing nutrients by african grazers

Experiments in Serengeti National Park, Tanzania, provide direct evidence that large, free-ranging mammalian grazers accelerate nutrient cycling in a natural ecosystem in a way that enhances their own carrying capacity. Both nitrogen and sodium were at considerably higher plant-available levels in soils of highly grazed sites than in soils of nearby areas where animal density is sparse. Fencing that uncoupled grazers and soils indicated that the animals promote nitrogen availability on soils of inherently similar fertility and select sites of higher sodium availability as well as enhancing that availability.     

Evaluation of Fertilizing Potential of Frozen-thawed dog Spermatozoa Diluted in ACP-106® using an In Vitro Sperm–Oocyte Interaction Assay

The aim of present study was to evaluate frozen canine semen with ACP-106 (Powder Coconut Water) using an in vitro sperm--oocyte interaction assay (SOIA). Ten ejaculates from five stud dogs were diluted in ACP-106 containing 20% egg yolk, submitted to cooling in a thermal box for 40 min and in a refrigerator for 30 min. After this period, a second dilution was performed using ACP-106 containing 20% egg yolk and 12% glycerol. Samples were thawed at 38 degrees C for 1 min. Post-thaw motility was evaluated by light microscopy and by using a computer aided semen analysis (CASA). Plasma membrane integrity and sperm morphology/acrosomal status were evaluated by fluorescent probes (C-FDA/PI) and Bengal Rose respectively. Moreover, frozen-thawed semen was analysed by a SOIA. Subjective post-thaw motility was 52.0 +/- 14.8% and it was significant higher than the total motility estimated by CASA (23.0 +/- 14.8%) because this system considered the egg yolk debris as immotile spermatozoa. Although normal sperm rate and acrosomal integrity evaluated by Bengal Rose stain was 89.6 +/- 3.1% and 94.3 +/- 3.1%, respectively, post-thaw percentage of intact plasma membrane was only 35.1 +/- 14.3%. Regarding SOIA, the percentage of interacted oocytes (bound, penetrated and bound and/or penetrated) was 75.3%. Using regression analysis, it was found significant relations between some CASA patterns and data for SOIA. In conclusion, the freezing-thawing procedure using ACP-106 was efficient for maintain the in vitro fertility potential of dog spermatozoa.     

Epigenetic Programming of Porcine Endometrial Function and the Lactocrine Hypothesis

Epigenetic programs controlling development of the female reproductive tract (FRT) are influenced by the effects of naturally occurring bioactive agents on patterns of gene expression in FRT tissues during organizationally critical periods of foetal and perinatal life. Aberrations in such important cellular and molecular events, as may occur with exposure to natural or manmade steroid or peptide receptor-modulating agents, disrupt the developmental program and can change the developmental trajectory of FRT tissues, including the endometrium, with lasting consequences. In the pig, as in other mammals, maternal programming of FRT development begins pre-natally and is completed post-natally, when maternal effects on development can be communicated via signals transmitted in milk. Studies involving relaxin (RLX), a prototypic milk-borne morphoregulatory factor (MbF), serve as the basis for ongoing efforts to identify maternal programming events that affect uterine and cervical tissues in the neonatal pig. Data support the lactocrine hypothesis for delivery of MbFs to neonates as a specific consequence of nursing. Components of a maternally driven lactocrine mechanism for RLX-mediated signalling in neonatal FRT tissues, including evidence that milk-borne RLX is delivered into the neonatal circulation where it can act on RLX receptor (RXFP1) -positive neonatal tissues to affect their development, are in place in the pig. The fact that all newborn mammals drink milk extends the timeframe of maternal influence on neonatal development across many species. Thus, lactocrine transmission of milk-borne developmental signals is an element of the maternal epigenetic programming equation that deserves further study.