Cloned cattle derived from a novel zona-free embryo reconstruction system

As the demand for cloned embryos and offspring increases, the need arises for the development of nuclear transfer procedures that are improved in both efficiency and ease of operation. Here, we describe a novel zona-free cloning method that doubles the throughput in cloned bovine embryo production over current procedures and generates viable offspring with the same efficiency. Elements of the procedure include zona-free enucleation without a holding pipette, automated fusion of 5-10 oocyte-donor cell pairs and microdrop in vitro culture. Using this system, zona-free embryos were reconstructed from five independent primary cell lines and cultured either singularly (single-IVC) or as aggregates of three (triple-IVC). Blastocysts of transferable quality were obtained at similar rates from zona-free single-IVC, triple-IVC, and control zona-intact embryos (33%, 25%, and 29%, respectively). In a direct comparison, there was no significant difference in development to live calves at term between single-IVC, triple-IVC, and zona-intact embryos derived from the same adult fibroblast line (10%, 13%, and 15%, respectively). This zona-free cloning method could be straightforward for users of conventional cloning procedures to adopt and may prove a simple, fast, and efficient alternative for nuclear cloning of other species as well

Multiple origins of hydrogenosomes:functional and phylogenetic evidence from the ADP/ATP carrier of the anaerobic chytrid Neocallimastix sp.

A mitochondrial-type ADP/ATP carrier (AAC) has been identified in the hydrogenosomes of the anaerobic chytridiomycete fungus Neocallimastix sp. L2. Biochemical and immunocytochemical studies revealed that this ADP/ATP carrier is an integral component of hydrogenosomal membranes. Expression of the corresponding cDNA in Escherichia coli confers the ability on the bacterial host to incorporate ADP at significantly higher rates than ATP - similar to isolated mitochondria of yeast and animals. Phylogenetic analysis of this AAC gene (hdgaac ) confirmed with high statistical support that the hydrogenosomal ADP/ATP carrier of Neocallimastix sp. L2 belongs to the family of veritable mitochondrial-type AACs. Hydrogenosome-bearing anaerobic ciliates possess clearly distinct mitochondrial-type AACs, whereas the potential hydrogenosomal carrier Hmp31 of the anaerobic flagellate Trichomonas vaginalis and its homologue from Trichomonas gallinae do not belong to this family of proteins. Also, phylogenetic analysis of genes encoding mitochondrial-type chaperonin 60 proteins (HSP 60) supports the conclusion that the hydrogenosomes of anaerobic chytrids and anaerobic ciliates had independent origins, although both of them arose from mitochondria

Global, regional, and national age-sex-specific mortality for 282 causes of death in 195 countries and territories, 1980-2017: a systematic analysis for the Global Burden of Disease Study 2017.

BACKGROUND: Global development goals increasingly rely on country-specific estimates for benchmarking a nation's progress. To meet this need, the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2016 estimated global, regional, national, and, for selected locations, subnational cause-specific mortality beginning in the year 1980. Here we report an update to that study, making use of newly available data and improved methods. GBD 2017 provides a comprehensive assessment of cause-specific mortality for 282 causes in 195 countries and territories from 1980 to 2017. METHODS: The causes of death database is composed of vital registration (VR), verbal autopsy (VA), registry, survey, police, and surveillance data. GBD 2017 added ten VA studies, 127 country-years of VR data, 502 cancer-registry country-years, and an additional surveillance country-year. Expansions of the GBD cause of death hierarchy resulted in 18 additional causes estimated for GBD 2017. Newly available data led to subnational estimates for five additional countries-Ethiopia, Iran, New Zealand, Norway, and Russia. Deaths assigned International Classification of Diseases (ICD) codes for non-specific, implausible, or intermediate causes of death were reassigned to underlying causes by redistribution algorithms that were incorporated into uncertainty estimation. We used statistical modelling tools developed for GBD, including the Cause of Death Ensemble model (CODEm), to generate cause fractions and cause-specific death rates for each location, year, age, and sex. Instead of using UN estimates as in previous versions, GBD 2017 independently estimated population size and fertility rate for all locations. Years of life lost (YLLs) were then calculated as the sum of each death multiplied by the standard life expectancy at each age. All rates reported here are age-standardised

Nondestructive determination of nutritional information in oilseed rape leaves using visible/near infrared spectroscopy and multivariate calibrations

Nitrogen, phosphorus and potassium content are the three most important nutritional parameters for growing oilseed rape. We investigated visible and near infrared (Vis/NIR) spectroscopy combined with chemometrics for the fast and nondestructive determination of nutritional information in oilseed rape leaves. A total of 154 leaf samples were collected, with 104 randomly selected as the calibration set, and the remaining 50 samples used as the validation set. The performance of eight different preprocessing methods was compared in partial least squares (PLS) models. Some effective wavelengths selected by a successive projections algorithm (SPA) were also used to develop linear SPA-PLS, nonlinear back propagation neural network (BPNN), and nonlinear least squares-support vector machine (LS-SVM) models to determine nutritional information. The best prediction models were DOSC-PLS for nitrogen with r=0.9743 and RMSEP=0.1459, DOSC-SPA-BPNN for phosphorus with r=0.7054 and RMSEP=0.0594, and DOSC-SPA-BPNN for potassium with r=0.9380 and RMSEP=0.1788. The prediction precision for nitrogen and potassium determinations was acceptable for further practical applications, whereas further studies are needed to improve the prediction precision for phosphorus. The results indicated that Vis/NIR spectroscopy is feasible for nondestructive determination of nutritional information in oilseed rape leaves. It also provided an alternative technique for detecting other growth information about oilseed rape leaves

Influence of breastmilk on the development of resistance to intestinal colonization in infants born at the Atma Java Hospital, Jakarta

A study of intestinal colonization resistance (CR) in breastfed versus formula-fed newborns at 4 intervals after birth in Jakarta, Indonesia, is described. To measure the intestinal CR for gram-negative enterobacilli, mean values of Enterobacteriaceae concentrations and mean numbers of Enterobacteriaceae biotypes were determined. The CR values found in this study show, that in all 4 sampling periods, at <1, 2, 4 and 6 months, the mean concentration of Enterobacteriaceae was somewhat lower in the breastfed group than in the formula-fed group (only significant at 6 months). This means that the intestinal CR of the breastfed group may have been slightly higher than that in the formula-fed group. In both study groups, the CR was lower in the second and fourth month than soon after birth and at 6 months. For epidemiological reasons, comparison was performed of the Enterobacteriaceae biotypes found in samples from mother and child. The data show that, in the first sampling period, regardless of the theoretical possibility of a 'more intense (skin) contact' during breastfeeding (which might promote transfer of also microorganisms), the breastfed infants had a significantly lower percentage of identical Enterobacteriaceae biotypes than did the formula-fed group. This could possibly be ascribed to a higher CR in the breastfed group. Determination of the concentration of Enterococcus species was found applicable to reproducably measure the CR in the newborns at 6 months and in the mother-group