Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Background  

The establishment of mutant populations together with the strategies for targeted mutation detection has been applied successfully to a large number of organisms including many species in the plant kingdom. Considerable efforts have been invested into research on tomato as a model for berry-fruit plants. With the progress of the tomato sequencing project, reverse genetics becomes an obvious and achievable goal.     

Reproductive Performance and Reconception of Nellore Cows According to Their Pure‐ or Cross‐Bred Calves

The objective of the present research was to evaluate the effect of a calf's genetic group on the productive and reproductive efficiency of its Nellore dam. Fixed‐time artificial insemination was applied to 800 cows using semen extracted from Nellore, Simmental and Angus Red bulls. Four hundred eleven cows calved, producing 119 Nellore, 103 ½Simmental–½Nellore and 189 ½Nellore–½Angus Red calves. The second mating period, which paired Nellore cows with Simmental bulls, was initiated 10 days after parturitions began and lasted for 5 months. Based on the two successive parturitions, the cumulative parturition rate for calving periods of 3, 4 and 5 months was calculated. Although no significant difference was observed for birth weight among the genetic groups, cross‐bred calves weighed, on average, 10% more than did pure‐bred calves at the age of 205 days. Nellore dams experienced a gestation period that was 7 days longer than did the cross‐bred dams, and the former showed a higher parturition rate at 90 and 120 days of the calving season, but not at 150 days (calving rates of 80.6, 76.4 and 76.2% for mothers of Nellore, ½Nellore–½Angus Red and ½Nellore–½Simmental, respectively, p > 0.05). At 90 and 120 days, Nellore dams produced more kg of calf per mated dam. In conclusion, in a short breeding season, Nellore dams nursing pure‐bred Nellore calves were found to have a higher biological efficiency compared with Nellore dams nursing cross‐bred calves.     

Anhydrous goat's milk fat: thermal and structural behaviors studied by coupled differential scanning calorimetry and X-ray diffraction. 2. Influence of cooling rate

Crystallization and melting properties of triacylglycerols (TGs) in anhydrous goat's milk fat (AGMF) are investigated by X-ray diffraction as a function of temperature (XRDT) coupled with high-sensitivity differential scanning calorimetry (DSC), using synchrotron radiation and Microcalix. The polymorphic behavior of AGMF was monitored by varying the cooling rates between 5 and 1 degrees C/min from 45 to -20 degrees C with their subsequent melting at 1 degrees C/min. Quenching of AGMF at -20 degrees C was also examined to determine the metastable polymorphic form of AGMF. At intermediate cooling rates, TGs in AGMF crystallize, from about 18 degrees C in two different lamellar structures with triple chain length 3Lalpha stacking of 72 A and a double chain length 2Lalpha stacking of 48 A, which are correlated to two overlapped exothermic peaks recorded by DSC. A reversible transition sub alpha <--> alpha was observed. Subsequent heating at 1 degrees C/min shows numerous structural rearrangements before final melting. At fast cooling of AGMF (5 degrees C/min), similar unstable crystalline varieties are formed while three endotherms are recorded. Several new unstable lamellar structures are observed after quenching. All of these data are compared to those previously reported at slow cooling (0.1 degrees C/min) showing a relative stability of the structures formed. In spite of general similitude, the thermal and structural behavior of the goat's milk is more complex than that of the cow's milk.     

Physiological and biochemical consequences of electroimobilisation in conscious sheep

An electroimmobilisation device has been developed to facilitate the automated shearing of sheep, but there is little information on its effects on the body. We have studied its effects on the cardiovascular system and on intermediary metabolism in sheep. Electroimmobilisation caused statistically significant increases in mean arterial pressure, heart rate, cardiac output, renal and hepatic and hindquarter glucose and lactate flux, organ and whole body oxygen flux, hindquarter blood flow and core temperature and decreases in arterial and posterior vena cava blood pH, renal and hepatic blood flow and PaCO2. Notably, no change occurred in PaO2. The metabolic changes demonstrated the capacity of sheep to respond to the increased muscular and cardiovascular work induced by electroimmobilisation. Pulmonary function was not compromised during electroimmobilisation as judged from blood gas changes, and the acid/base changes were rapidly reversed after electroimmobilisation. The recovery to control conditions for all perturbations generally took no longer than 30 min, consistent with a rapid and physiologically adequate reversal by the animal's homeostatic mechanisms.     

Imaging of intermittency in ripple-wave turbulence

The dynamics of a fluid surface filled with high-amplitude ripples were studied with a technique (diffusing light photography) that resolves the height at all locations instantaneously. Even when nonlinearities are strong enough to generate a (Kolmogorov) cascade from long wavelength (where energy is input) to shorter wavelength, the resulting turbulent state contains large coherent spatial structures. The appearance of these structures in a thermal equilibrium state (with the same average energy) would be highly improbable.     

Effect of Chemical or Electrical Activation of Bovine Oocytes on Blastocyst Development and Quality

Activation of in vitro‐matured (IVM) oocytes is essential for successful embryo production following nuclear transfer (NT) or intracytoplasmic sperm injection (ICSI). This study was designed to compare the rates of blastocyst production and embryo quality (as measured by numbers of viable cells) following parthenogenetic activation with electrical pulse or the use of two different calcium ionophores, A23187 (CA) or ionomycin (IO), with or without the addition of bovine serum albumin (BSA). IVM oocytes with a first polar body were randomly allocated to five treatment groups: CA (5 μm CA, 5 min; n = 88), CA + BSA (5 μm CA, 5 min; BSA, 5 min; n = 90), IO (5 μm IO, 5 min; n = 91), IO + BSA (5 μm IO, 5 min; BSA, 5 min; n = 86) and EL (two pulses of 1.5 kV/cm, 20 μs; n = 120). Blastocyst rates were higher (p < 0.05) for CA (54.4%), IO (51.4%) and EL (54.5%) than for IO + BSA (18.3%). Treatment CA + BSA (39.8%) did not differ from the others. There was no difference (p > 0.05) among treatments in total number of cells. However, the percentage of viable cells was reduced in CA (49.9%), CA + BSA (45.8%), IO (64.9%), IO + BSA (50.9%) compared with EL (82.7%). In summary, the addition of BSA to the IO treatment had an adverse effect on blastocyst production rates. Although there was no difference between electrical stimulation and chemical activation on blastocyst production rates, electrical activation resulted in blastocysts with a higher percentage of viable cells.     

Anhydrous goat's milk fat: thermal and structural behavior. 1. Crystalline forms obtained by slow cooling

The thermal and structural behaviors of anhydrous goat's milk fat (AGMF) have been determined as a function of temperature using a powerful technique allowing simultaneous time-resolved synchrotron X-ray diffraction as a function of temperature (XRDT) and high-sensivity differential scanning calorimetry (DSC) measurements from the same sample. This first paper, aiming at the characterization of the physical properties of AGMF, we examine crystalline organizations made by triacylglycerols (TG) upon slow cooling at /dT/dt/ = 0.1 degrees C/min from 45 to -20 degrees C in order to approach system equilibrium. Three overlapped exotherms were observed by DSC upon cooling, whereas four endotherms were found on the subsequent heating at 1 degrees C/min. XRDT evidenced that AGMF crystallizes under four different lamellar structures, two with double-chain length packings at 41.5 and 38.2 angstroms and two with triple-chain lengths of 72 and 64.7 angstroms stacking. Simultaneous wide-angle XRDT has shown that initial nucleation mainly occurs in a packing of beta' type from approximately 26 degrees C, although some transient presence of alpha was detected. The absence of polymorphic transition, on heating, until final melting (approximately 40 degrees C) demonstrated the relative stability of the structures formed.     

Vitrification with Glutamine Improves Maturation Rate of Vitrified / Warmed Immature Bovine Oocytes

The current study examined the protective effects of l ‐glutamine and cytochalasin B during vitrification of immature bovine oocytes. Oocyte vitrification solution (PBS supplemented with 10% FCS, 25% EG, 25% DMSO and 0.5 m trehalose) was the vitrification control. Treatments were the addition of 7 μg/ml cytochalasin B, 80 mm glutamine or both cytochalasin and glutaminine for 30 s. After warming, oocytes were matured in vitro for 24 h, fixed and stained with Hoechst (33342) for nuclear maturation evaluation. l ‐glutamine improved the vitrified/warmed immature bovine oocytes viability (32.8%), increasing the nuclear maturation rates compared to other treatments and the no treatment vitrified control (17.4%). There was, however, no effect of cytochalasin B on in vitro maturation (14.4%).     

Crystallization properties and polymorphism of triacylglycerols in goat's milk fat globules

The sensorial, functional, and nutritional properties of goat dairy products result from the specific fatty acid composition of goat's milk fat. However, information on the physical and thermal properties of goat's milk fat is scarce. In this study, crystallization of triacylglycerols (TG) in goat's milk fat globules was investigated using polarized light microscopy and the coupling of time-resolved synchrotron radiation X-ray diffraction (XRD) and high-sensitivity differential scanning calorimetry (DSC). The molecular organization of the solid fat phase was characterized for cooling rates between 3 and 0.1 degrees C/min. Quenching of goat's milk fat globules from 50 to -8 degrees C and 4 degrees C was also examined to identify the most unstable polymorphic forms of TG. Then, the melting behavior of fat crystals was studied on subsequent heating at 1 degrees C/min. Triple chain length (3L: 68.6-70 A) and double chain length (2L: 37-45.4 A) structures were characterized and 5 polymorphic forms, alpha, sub-alpha, beta' 1, beta' 2, and beta were identified. Polymorphic transitions were observed within goat's milk fat globules as a function of time after quenching and as a function of temperature on heating. From a technological point of view, this work will contribute to a better understanding of the rheological properties as well as on the flavor evolutions of goat's milk-based products.