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The aqueous self-assembly of methyl-terminated tri(oxyethylene)thiol-encapsulated gold nanoclusters of varying core size is demonstrated on micrometer scale Au/SiO2 interdigital electrodes. This self-assembly process consists of alternate exposures of the substrate to solutions of either an α,ω-dithiol or the gold nanoclusters, resulting in the deposition of these materials onto the electrode surface. A comparison of the procedure in both H2O and CHCl3 solvents shows that the assembly, as monitored by the electrical conductivity of the device, occurs more rapidly in the H2O system. This observation is complimented by XPS and UV/Vis measurements, which show that (1) the increased current is due to an increased amount of gold deposited on the surface under aqueous conditions and (2) the thiol exchange reaction occurs more rapidly in H2O in comparison to CHCl3.

The storage and behavior of sperm collected from the epididymis of two emyiid turtles were examined. In Chrysemys picta, the weight of the epididymis does not change significantly throughout the year as does the testis. However, in this species, as well as in Trachemys scripta, the epididymis contains sperm throughout the entire year. Sperm from both species have a relatively low motility and velocity of movement. In C. picta, equally low motilities are observed both in the autumn, shortly after spermiation, and in spring. Motility could be augmented by the addition of isobutyl methyl xanthine (IBMX, 0.5 mmol l^–1). Epididymal sperm remained viable in excess of 40 days in vitro when stored in F-10 buffer, during which time motility and swimming velocity could be augmented with IBMX. The longevity and low motility of turtle sperm facilitates its storage by either males or females, and creates conditions that promote the wide dissemination, over time, of gametes produced in a narrow time window. J. Exp. Zool. 292:180–186, 2002. © 2002 Wiley-Liss, Inc.

Optimization of culture techniques for turtle and other reptilian lymphocytes is essential for facilitating cytogenetic and immunologic research for these animals. We examined a variety of conditions and parameters relevant to turtle lymphocyte culture including: different mitogenic agents, alone and in combination; lymphocyte separation protocols; culture volume; time required to stimulate lymphocytes to mitosis; importance of humidity and gas exchange in culture incubation; suitability of different culture media; effects of varying serum concentrations; ability of interleukin-2 (IL-2) to stimulate lymphocyte growth and prevent apoptosis; and feasibility of inducing premature chromosome condensation. The best conditions of those we studied for obtaining mitotic cells were (1) the combined use of phytohemagglutinin-M form (2%) and lipopolysaccharides (0.55 µg/ml), (2) the use of 5% autologous turtle serum (as opposed to fetal bovine serum), and (3) collection of mitotic cells around 96 hours after mitogenic stimulation. Human, recombinant IL-2 did not increase the fraction of lymphocytes in mitosis over the range of concentrations tested and calyculin A was ineffective at inducing premature chromosome condensation in turtle lymphocytes over the range of concentrations tested. This test regime provides a guideline for determination of appropriate lymphocyte culture conditions in turtles and other reptiles.

Some freshwater turtles appear unable to produce eggs large enough to achieve the balance between size and number of eggs predicted by optimal egg size theory. We present evidence that pelvic girdle structure constrains egg size and thus offspring size in females of smaller-bodied species (Chrysemys picta and Deirochelys reticularia). The constraint is demonstrated by the correspondence of slopes of the increase of the pelvic aperture and egg width with increasing body size. This constraint appears to be relaxed in a larger-bodied species (Pseudemys scripta), in which the increase in pelvic aperture relative to body size is greater than the increase in egg width. This type of structural constraint on a reproductive trait should not occur unless there is strong selection on pelvic architecture for other functions, such as locomotion, support, and limb retraction, that prevent expansion of the pelvic aperture. Although other explanations may exist for other groups of organisms that appear to vary egg size, the large variation in egg size associated with body size in some species of turtles can be reconciled with optimal egg size theory only if a pelvic constraint model is accepted.