Extrachromosomal microDNAs and chromosomal microdeletions in normal tissues

We have identified tens of thousands of short extrachromosomal circular DNAs (microDNA) in mouse tissues as well as mouse and human cell lines. These microDNAs are 200 to 400 base pairs long, are derived from unique nonrepetitive sequence, and are enriched in the 5'-untranslated regions of genes, exons, and CpG islands. Chromosomal loci that are enriched sources of microDNA in the adult brain are somatically mosaic for microdeletions that appear to arise from the excision of microDNAs. Germline microdeletions identified by the "Thousand Genomes" project may also arise from the excision of microDNAs in the germline lineage. We have thus identified a previously unknown DNA entity in mammalian cells and provide evidence that their generation leaves behind deletions in different genomic loci.     

Diversity in starch, protein and mineral composition of sorghum landrace accessions from Ethiopia

Sorghum (Sorghum bicolor (L.) Moench) is an important food crop in arid and semi-arid tropics where it is grown mainly due to its drought tolerance. In Ethiopia, sorghum is grown in different agro-ecological zones primarily for food. Landraces are widely cultivated in the region due to their adaptation to the growing conditions and other characteristics such as food quality, quantity, and quality of by-products. The objective of this research was to determine the genetic variation in protein, mineral composition, total starch and its components in sorghum landraces collected from the western regions of Ethiopia. Sorghum whole grains were analyzed for crude protein, total starch and its components, and mineral profile (calcium, magnesium, potassium, phosphorus, iron, manganese, zinc, and sodium). The analysis of variance showed significant differences among the sorghum landraces in nutritional composition. There was highly significant correlation between phosphorus and magnesium, and between zinc and phosphorus and protein which indicated that some interaction existed in absorption and translocation between minerals and protein content. The genetic variation found among the sorghum landrace accessions in chemical composition can be exploited in the breeding program.     

The Effects of Frequent Electroejaculation on the Semen Characteristics of a Captive Siberian Tiger (Panthera tigris altaica)

Artificial insemination (AI) can help to avoid inbreeding and genetic degeneration for sustaining genetically healthy populations of endangered species in captivity. Collection of a sufficient quantity of viable sperm is an essential first step in the AI process. In the present study, we examined the effects of frequent electroejaculation on semen characteristics in a Siberian tiger. We collected semen in all 17 trials during 6 breeding seasons (6 years). The mean number of sperm and the percentage of motile sperm were 294.3 ± 250.2×10⁶/ejaculate and 82.4 ± 11.4%, respectively. The number of motile sperm tended to increase during frequent electroejaculation in the same breeding season. Semen collection by electroejaculation can be performed effectively up to the fourth sequential ejaculate, which contained the most sperm in the study. In conclusion, frequent collection of sperm by electroejaculation from tigers may be effective for collection of a large number of motile sperm.     

Preparation of liquid polyesterpolyols from glucose and its methyl derivative

Preparation of liquid polyols from d-(+)-glucose (Glc) and its derivative, methyl--d-glucoside (m-Glc), has been studied. Direct reaction of -caprolactone (CL) with Glc (CL:Glc = 2:1–5:1 in weight ratio) at 150°C using tin(II) 2-ethyl hexanoate (SnEht₂) (series A), in which melted Glc was suspended in CL, resulted in a dark-brown coloration of the reactants. The reaction was accompanied by formation of high molecular weight resins, a pH drop, production of water, and a considerable decrease in the hydroxyl value from the theoretically expected one. In the case of Glc/ethylene glycol (EG)/CL reaction system with SnEht₂ catalyst at 150°C (series B), in which the weight ratio of Glc to EG was fixed at 1:1, Glc dissolved in the EG/CL mixture, but the brown coloration of the reactant mixture still occurred. In this case, the formation of water was enhanced, but the other effects found in series A were suppressed to a considerable extent. In the m-Glc/CL/SnEht₂ reaction system (series C), in which m-Glc reacted with two to five times weight amounts of CL under the same conditions adopted in series A, development of the color, the production of high molecular weight materials and water, and the changes in pH and hydroxyl value were not observed. These results are discussed based on the chemical structural differences: Glc exists mostly in the hemiacetal form, but tautomerizes to the aldehyde, whereas m-Glc is an acetal and is protected from reversion to the aldehyde.     

Agronomic evaluation and identification of potential cowpea (Vigna unguiculata L. Walp) genotypes in South Africa

Cultivar development of cowpea with increased grain yield depends on evaluation and selection of genetically unique and complementary breeding lines. The objective of this study was to evaluate cowpea collections using agronomic traits to select promising parents to develop breeding population. Twenty-two cowpea genotypes were field evaluated using a randomised complete block design with three replications in two locations in South Africa. Analysis of variance revealed significant genotypic (G) and locations (L) differences for all agronomic traits evaluated. Significant differences were also observed for most of the traits due to G X L, G X Year (Y), L X Y and G X L X Y interactions effects. The genotypes tested at two locations showed considerable genetic variation for the following agronomic traits that varied for number of leaves (NL) (20.08–179.78), leaf length (LL) (12.36–67.72 cm), leaf width (LW) (4.78–22.02 cm), number of branches (NB) (6.81–13.85), number of pods per plant (NPPP) (14.33–54.04), number of seeds per pod (NSPP) (11.7286.28), pod diameter (PD) (5.75–22.73 mm), plant height (PH) (6.57–233.71 cm), pod length (PL) (3.94–100.92 mm), hundred seed weight (HSWt) (8.11–49.52 g) and grain yield (GY) (0.10-1.48 t ha^?1). Grain yield was significant and positively correlated with all of the traits except leaf length. The principal component analysis (PCA) identified four principal components (PCs) contributing to 73.62% of the total explained variation amongst the tested genotypes. The selected cowpea genotypes for most of agronomic characteristics are useful and candidate genetic resources for the development of breeding population in cowpea breeding and enhanced production and productivity for food and nutrition in the country.     

Preimplantation development of embryos in labrador retrievers

Preimplantation development of canine embryos is not well understood. To understand the timing of preattachment embryogenesis relative to the luteinizing hormone (LH) surge, early embryonic development was examined in Labrador Retrievers after artificial insemination. The embryos migrated from the oviduct to the uterus beginning on day 11 after the LH surge. This transport must be completed within 24 h. By day 13 after the LH surge, all of the embryos had moved and were localized in the uterus. The embryos developed to the morula stage within 11-13 days and to the blastocyst stage within 14 days after the LH surge, respectively. These findings add to the current understanding concerning the physiology of preimplantation development and should help further develop assisted reproductive techniques in canine species, such as cryopreservation and subsequent embryo transfer.     

Effects of resveratrol treatment on mitochondria and subsequent embryonic development of bovine blastocysts cryopreserved by slow freezing

This study evaluated the effects of cryopreservation by slow freezing on the mitochondrial function, DNA integrity, and developmental ability of bovine embryos and examined whether resveratrol treatment of the frozen‐thawed blastocysts improved embryonic viability. In vitro produced bovine embryos were subjected to slow freezing. After thawing, the ATP content and mitochondrial DNA integrity (mtDNA), determined by real‐time PCR targeting short and long mitochondrial sequences, was found to be lower in frozen‐thawed embryos than in fresh embryos, and mtDNA copy number was significantly reduced during the 24‐hr incubation post warming. Furthermore, immunostaining against double‐strand DNA revealed DNA damage in frozen‐thawed embryos. When frozen‐thawed embryos were incubated in the medium containing 0.5 µM resveratrol, SIRT1 expression, and survival rate of the embryos significantly improved compared with the vehicle‐treated embryos. In addition, cell‐free mtDNA content in medium was higher in case of resveratrol‐treated embryos than of vehicle‐treated embryos. In conclusion, slow freezing affects mitochondrial integrity and function in the blastocysts. In the frozen‐thawed embryos, mitochondria were removed during post‐thawing incubation and resveratrol enhanced the process, resulting in improved survivability of the embryos.