Use of Density Centrifugation for Delayed Cryopreservation of Stallion Sperm: Perform Sperm Selection Directly after Collection or after Storage?

Equipment for cryopreservation of stallion sperm is not always available. In such cases, diluted semen can be shipped to a facility for later cryopreservation. The aim of this study was to evaluate if selection of sperm via density centrifugation yields higher survival rates when cryopreservation is to be delayed (i.e. carried out after 1 day of storage at 5°C). Two‐layer iodixanol as well as single‐layer Androcoll density centrifugation were tested and compared with samples prepared with standard centrifugation. Special emphasis was placed on comparing centrifugation on the day of semen collection with centrifugation after 1‐day refrigerated storage. Sperm morphology and motility as well as membrane and chromatin integrity were evaluated before and after centrifugation. Sperm motility and membrane integrity were also assessed after cryopreservation. It was found that both two‐ and single‐layer density centrifugation processing resulted in higher percentages of morphologically normal and motile sperm with higher membrane and chromatin integrity, as compared to standard centrifugation or diluted samples. Differences were only in the order of magnitude of 5%. Recovery rates after density centrifugation were only approximately 30–40%. When cryopreservation was carried out after 1‐day refrigerated storage, centrifugation processing of sperm directly after semen collection resulted in higher percentages of plasma membrane intact sperm post‐thaw as compared to performing centrifugation processing of stored sperm just prior to cryopreservation. No significant differences in progressively motile sperm post‐thaw were seen. Taken together, for delayed cryopreservation, it is best to perform density centrifugation directly after collection rather than immediately prior to cryopreservation.     

Fine-Scale Doppler Radar Observations of Tornadoes

Observations obtained with a mobile pencil-beam Doppler radar revealed many previously unresolved structures within tornadic storms and tornadoes and helped verify various aspects of conceptual models. Radar data from the parent circulations indicate the existence of spiral reflectivity bands, intense radial wind shear zones, and multiple larger-scale velocity maxima. Tornado structures observed include debris shields, clear axial (eye) regions, multiple reflectivity bands surrounding the center of the eye, and occasional reflectivity protrusions into the eye. Velocity and reflectivity data from tornado-scale circulations show evidence of axial downdrafts.     

Spectral Properties of Near-Earth Asteroids: Evidence for Sources of Ordinary Chondrite Meteorites

Although ordinary chondrite (OC) meteorites dominate observed falls, the identification of near-Earth and main-belt asteroid sources has remained elusive. Telescopic measurements of 35 near-Earth asteroids ( approximately3 kilometers in diameter) revealed six that have visible wavelength spectra similar to laboratory spectra of OC meteorites. Near-Earth asteroids were found to have spectral properties that span the range between the previously separated domains of OC meteorites and the most common (S class) asteroids, suggesting a link. This range of spectral properties could arise through a diversity of mineralogies and regolith particle sizes, as well as through a time-dependent surface weathering process.     

Thermally Switchable Periodicities and Diffraction from Mesoscopically Ordered Materials

Two switchable, mesoscopically periodic materials were created by combining crystalline colloidal array (CCA) self-assembly with the temperature-induced volume phase transition of poly(N-isopropylacrylamide) (PNIPAM). Body-centered-cubic CCAs of hydrated, swollen PNIPAM particles Bragg-diffract infrared, visible, and ultraviolet light weakly, whereas arrays of compact shrunken particles diffract efficiently. A tunable diffracting array was also created by embedding a CCA of polystyrene spheres within a PNIPAM hydrogel that swells and contracts with temperature; thus the array lattice constant varies with temperature, and the diffracted wavelength was thermally tunable across the entire visible spectrum. These materials may find applications in many areas of optics and materials science.     

Genetic structure of Leptopilina boulardi populations from different climatic zones of Iran

Background  

The genetic structure of populations can be influenced by geographic isolation (including physical distance) and ecology. We examined these effects in Leptopilina boulardi, a parasitoid of Drosophila of African origin and widely distributed over temperate and (sub) tropical climates.     

24-Hour Secretion Patterns of Plasma Oestradiol 17β in Pony Mares in Late Gestation

The mare exhibits nocturnal uterine contractions in the last 6 days of gestation. It is hypothesized that estradiol 17β (O17β) may be associated with the nightly increase in uterine contractions. The 24‐h secretion pattern of plasma O17β was measured in 3 pony mares in late gestation to identify changes in release as the mare neared parturition. Blood was collected weekly at 08:00 hours beginning on day 240 and every third day from day 330 until delivery. Serial blood samples were collected from each mare every 30‐min for 24‐h beginning on gestation day 310 and every sixth day thereafter until parturition. Concentrations of O17β were elevated at night with lowest concentrations occurring directly before sunset (p < 0.01). The natural log of the variance was increased at sunset (p < 0.01) and was decreased during the 6‐h period immediately after sunrise. This pattern was especially evident in the 6 days that preceded parturition. The contrast between nocturnal and daytime concentrations of O17β in the last 6 days of gestation may contribute to night‐time delivery in the mare.     

The Effects of Hoechst 33342 Staining and the Male Sample Donor on the Sorting Efficiency of Canine Spermatozoa

The aim of this study was to evaluate the influence of Hoechst 33342 (H‐42) concentration and of the male donor on the efficiency of sex‐sorting procedure in canine spermatozoa. Semen samples from six dogs (three ejaculates/dog) were diluted to 100 × 10⁶ sperm/ml, split into four aliquots, stained with increasing H‐42 concentrations (5, 7.5, 10 and 12.5 μl, respectively) and sorted by flow cytometry. The rates of non‐viable (FDA+), oriented (OS) and selected spermatozoa (SS), as well as the average sorting rates (SR, sorted spermatozoa/s), were used to determine the sorting efficiency. The effects of the sorting procedure on the quality of sorted spermatozoa were evaluated in terms of total motility (TM), percentage of viable spermatozoa (spermatozoa with membrane and acrosomal integrity) and percentage of spermatozoa with reacted/damaged acrosomes. X‐ and Y‐chromosome‐bearing sperm populations were identified in all of the samples stained with 7.5, 10 and 12.5 μl of H‐42, while these two populations were only identified in 77.5% of samples stained with 5 μl. The values of OS, SS and SR were influenced by the male donor (p < 0.01) but not by the H‐42 concentration used. The quality of sorted sperm samples immediately after sorting was similar to that of fresh samples, while centrifugation resulted in significant reduction (p < 0.05) in TM and in the percentage of viable spermatozoa and a significant increase (p < 0.01) in the percentage of spermatozoa with damage/reacted acrosomes. In conclusion, the sex‐sorting of canine spermatozoa by flow cytometry can be performed successfully using H‐42 concentrations between 7.5 and 12.5 μl. The efficiency of the sorting procedure varies based on the dog from which the sperm sample derives.     

Effect of Follicle Size on In Vitro Maturation of Pre‐Pubertal Porcine Cumulus Oocyte Complexes

Very small follicles (<3.0 mm diameter) are over‐represented on the surface of ovaries of non‐cycling pigs, and the oocytes collected from these follicles generally have reduced developmental competence in vitro. This study examined the effect of follicle size on the nuclear maturation (n = 608), the potential of parthenogenetic activation (n = 243) and the cyclic AMP (cAMP) content of pre‐pubertal porcine oocytes (n = 480). In addition, the influence of follicle size on steroid hormone synthesis was analysed. Cumulus oocyte complexes (COCs) flushed from small (2.5–4.0 mm) or large (4.5–6.0 mm) ovarian follicles were cultured for 0, 28 and 46 h. After 46 h of IVM, a greater proportion of oocytes from 4.5‐ to 6.0‐mm follicles reach metaphase II (MII) compared with those from follicles with 2.5–4.0 mm of diameter (96.1 vs 77.0%, respectively; p < 0.001). Parthenogenetic activation of oocytes from large follicles produced higher developmental rates than oocytes from large follicles (p < 0.05). At 28 h, the IVM medium with oocytes from large follicles contained significantly more 17ß‐oestradiol (E₂) than the medium with oocytes from small follicles (5.55 vs 3.45 ng/ml, respectively; p < 0.05) and at 46 h, the medium with oocytes from small follicles contained significantly more progesterone (P₄) than the medium with oocytes from large follicles (276.7 vs 108.2 ng/ml, respectively, p < 0.05). Porcine oocytes from large follicles have higher nuclear and cytoplasmic maturation capacities, but the differences did not appear to be cAMP‐mediated. Our findings also suggest that COCs from small follicles undergo more intensive luteinization than COCs from large follicles. The results show that oocytes from follicles with a diameter greater than 4.0 mm are more suitable for in vitro studies.     

Visualization of three-dimensional chaos

Most chaotic mixing experiments have been restricted to two-dimensional, time-periodic flows, and this has shaped advances in theory as well. A prototypical, bounded, three-dimensional flow with a moderate Reynolds number is presented; this system lends itself to detailed experimental observation and allows for high-precision computational inspection. The flow structure, captured by means of cuts with a laser sheet (experimental Poincare section), was visualized with the use of continuously injected fluorescent dye streams and revealed detailed chaotic structures with high-period islands.