In vitro development of IVF-derived bovine embryos following cytoplasmic microinjection for the episomal expression of the IGF2 gene

Important genomic imprinting changes usually occur following the in vitro production (IVP) of bovine embryos, especially in the imprinting pattern of components of the IGF system. This study aimed to evaluate the effects of a transient episomal overexpression of the IGF2 gene in bovine IVP embryos following embryo cytoplasmic microinjection (CMI) at the 1-cell stage on embryo survival, early and late developmental kinetics and morphological quality up to Day 7 of development. Selected cumulus–oocyte complexes (COCs) were matured and fertilized in vitro and subsequently segregated into six experimental groups: non-CMI control group and five CMI groups at increasing doses (0, 10, 20, 40 and 80 ng/μl) of a GFP vector built for the episomal expression of bovine IGF2. Zygote CMI was effective in delivering the expression vector into the ooplasm, irrespective of the groups, with 58% of positive GFP fluorescence in Day 7 blastocysts. Considering developmental rates and late embryo kinetics, the 10-ng/μl CMI vector dose promoted a lower blastocyst rate (10.4%), but for blastocysts at more advanced stages of development (93.0% blastocysts and expanded blastocysts), and higher number of cells (116.0 ± 3.0) than non-CMI controls (23.3%, 75.0% and 75.0 ± 6.8 were obtained, respectively). In conclusion, CMI at the 1-cell stage did not compromise subsequent in vitro development of surviving embryos, with the 10-ng/μl group demonstrating a possible growth-promoting effect of the IGF2 gene on embryo development, from the 1-cell to the blastocyst stage.     

Modeling the Interfacial Interactions between CrtS and CrtR from Xanthophyllomyces dendrorhous , a P450 System Involved in Astaxanthin Production

Xanthophyllomyces dendrorhous is a natural source of astaxanthin, a carotenoid widely used in the food industry. In this yeast, astaxanthin is synthesized from β-carotene by a cytochrome P450, CrtS, which depends on CrtR, the four-domain cytochrome P450 reductase (CPR). Although Saccharomyces cerevisiae has an endogenous CPR (ScCPR), expression of CrtS does not result in astaxanthin production unless it is coexpressed with CrtR. Assuming that CrtS could interact with the FMN-binding domain of either CrtR or ScCPR (XdFMNbd and ScFMNbd, respectively), the aim of this work was to identify possible interaction differences between these alternative complexes by protein modeling and short molecular dynamics simulations. Considering the recently proposed membrane orientation of a mammalian P450, our CrtS-CrtR model predicts that both N-terminal ends stand adjacent to the membrane plane, allowing their anchoring. Compared with the possible interface between CrtS and both FMNbd, the Xanthophyllomyces system appears to be stabilized by more saline bridges.     

Linalool from Lippia alba: study of the reproducibility of the essential oil profile and the enantiomeric purity

A new chemotype of the aromatic Verbenaceae species Lippia alba Mill. N. E. Br. from southeastern Brazil has recently been shown to have a high content of linalool in the leaf essential oil. Vegetative propagation of this chemotype was conducted at six different locations in Brazil, and the variation of the content and the optical purity of linalool in the oils were verified. Yields (0.6-0.9%, hydrodistillation), chemical composition, linalool content, and optical purity of the oils from all the plants were compared, using GC-FID, GC-MS, chiral chromatography, and retention index calculation. No plant exceeded the matrix in linalool content (46.5 to 90.7%), and the chemical profile of the oils was the same for all the samples. Purification of linalool to a content close to 100% was effected by vacuum distillation of the crude oil. Chiral analysis showed exclusively the presence of S-linalool in all the crude oils and in the distilled samples.     

Liming as an ameliorator of copper toxicity in black oat (Avena strigosa Schreb.)

This study aimed to assess the effect of liming as an ameliorator of copper (Cu) toxicity in black oats (Avena strigosa Schreb.) grown in soil with a sandy texture. The treatments were soil supplementation with increasing doses of lime (0.0, 0.5 and 1.0 g kg^?1) and two of Cu (0 and 50 mg kg^?1). Pre-germinated black oat seeds were sown for 30 days in the soil, subsequently, the plant height, root and shoot dry mass and concentration of total chlorophyll, chlorophylls a and b, carotenoids and total soluble carbohydrates and starch were analyzed. The dose of 50 mg kg^?1 soil Cu caused a toxic effect on black oat plant growth and development, affected the internal distribution of nutrients, decreased the leaf chlorophyll concentration and increased the leaf concentration of total soluble carbohydrates and starch. Increasing lime doses ameliorated the toxic effects of Cu.     

Generation of a Mixolab Profile After the Evaluation of the Functionality of Different Commercial Wheat Flours for Hot‐Press Tortilla Production

Refined wheat flours commercially produced by five different U.S. and Mexican wheat blends intended for tortilla production were tested for quality and then processed into tortillas through the hot‐press forming procedure. Tortilla‐making qualities of the flour samples were evaluated during dough handling, hot pressing, baking, and the first five days on the shelf at room temperature. The predominant variables that affected the flour tortilla performance were wet gluten content, alveograph W (220–303) and P/L (0.70–0.94) parameters, farinograph water absorption (57%) and stability (10.8–18.7 min), starch damage (5.43–6.71%), and size distribution curves (uniform particle distribution). Flours produced from a blend of Dark Northern Spring (80%) and Mexican Rayon (20%) wheat had the highest water absorption, and tortillas obtained from this blend showed the highest diameter and lowest thickness. The whitest and best textured tortillas were obtained from the flour milled from three hard types of Mexican wheat blend. A Mixolab profile was generated from the best tortilla flours, those produced by mills 3 and 4. The Mixolab profile showed that a good flour for hot‐press tortillas had a relatively lower absorption and short dough mix time compared with a bread flour and should have a significantly higher gluten compared with an all‐purpose flour. Compared with bread flour, the tortilla flour had higher retrogradation and viscosity values. The Mixolab profile proved to be a good preliminary test to evaluate flours for hot‐press tortillas.     

Carcass characteristics and meat quality of sheep fed alfalfa hay to replace Bermuda grass hay

Tropical Animal Health and Production - The objective of this study was to evaluate carcass characteristics, meat quality, and organ and viscera weight of forty male sheep, not castrated, initially...     

Nitrogen uptake by sago palm (Metroxylon sagu Rottb.) in the early growth stages

Previous trials have revealed variable responses of sago palm ( Metroxylon sagu Rottb.) to fertilizer application, particularly nitrogen (N). In the present study, we quantified the fertilizer use efficiency (FUE) of sago palm for the first time using ¹⁵N-labeled fertilizer in pot and field experiments. The pot experiment was conducted in Japan using a 2:1 mixture of sand to Philippine soil. The field experiment was conducted in Leyte in the Philippines. Both experiments consisted of three replicates in each of three treatments: control, ¹⁵N urea at 50 kg N ha⁻¹ and ¹⁵N urea at 100 kg N ha⁻¹. The N uptake of sago palm increased significantly, but inconsistently with increasing N application. The few instances of a significant increase in N uptake did not translate into significant improvements in growth parameters, except for the number of leaflets in the pot experiment. The FUE values for sago seedlings (< 6 months) in the pot experiment treated with 50 and 100 kg N ha⁻¹ were 10.5 and 13.2%, respectively, whereas for the 2-year-old sago palms in the field, the corresponding FUE values were 14.8 and 12.0%. The FUE values were similar at the two levels of N application in both experiments. Sago growth parameters appeared to be insensitive to N application, suggesting that the form of N and the timing of N fertilization are important factors for sago palms. Therefore, the use of N fertilizer in sago production can only be justified after determining and fully understanding the response of sago palm to N application.