Lectin-binding sites on the plasma membranes of rabbit spermatozoa: Changes in surface receptors during epididymal maturation and after ejaculation

Modifications in rabbit sperm plasma membranes during epididymal passage and after ejaculation were investigated by used of three lectins: concanavalin A (Con A); Ricinus communis I (RCA(I)); and wheat germ agglutinin (WGA). During sperm passage from caput to cauda epididymis, agglutination by WGA drastically decreased, and agglutination by RCA(I) slightly decreased, although agglutination by Con A remained approximately unchanged. After ejaculation, spermatozoa were agglutinated to a similar degree or slightly less by Con A, WGA, and RCA(I), compared to cauda epididymal spermatozoa. Ultrastructural examination of sperm lectin-binding sites with ferritin- lectin conjugates revealed differences in the densities of lectin receptors in various sperm regions, and changes in the same regions during epididymal passage and after ejaculation. Ferritin-RCA(I) showed abrupt changes in lectin site densities between acrosomal and postacrosomal regions of sperm heads. The relative amounts of ferritin-RCA(I) bound to heads of caput epididymal or ejaculated spermatozoa. Tail regions were labeled by ferritin RCA(I) almost equally on caput and cauda epididymal spermatozoa, but the middle-piece region of ejaculated spermatozoa was slightly more densely labeled than the principal-piece region, and these two regions on ejaculated spermatozoa were labeled less than on caput and cuada epididymal spermatozoa. Ferritin-WGA densely labeled the acrosomal region of caput epididymal spermatozoa, although labeling of cauda epidiymal spermatozoa was relatively sparse except in the apical area of the acrosomal region. Ejaculated spermatozoa bound only a few molecules of ferritin-WGA, even at the highest conjugate concentrations used. Caput epididymal, but not cauda epididymal or ejaculated spermatozoa, bound ferritin-WGA in the tail regions. Dramatic differences in labeling densities during epididymal passage and after ejaculation were not found with ferritin-Con A.     

Estimating Duration of Short-Term Acute Effects of Capture Handling and Radiomarking

ABSTRACT Radiotelemetry is used extensively in zoographic studies of wildlife species, including northern bobwhite (Colinus virginianus). These studies assume that radiomarking does not affect survival of marked individuals. However, most researchers implicitly acknowledge that capture, handling, and radiomarking may have short-term deleterious effects on individuals and, therefore, include in analyses only animals that survive an adjustment period of arbitrary length (often 7 days) following capture and marking. Length of adjustment period is rarely empirically based and may potentially bias survival estimates. We outline an analytical approach to determine an appropriate adjustment period and illustrate this approach by examining effects of time-since-marking on survival of 410 northern bobwhite captured during winter from 1997 to 2001, in Mississippi, USA. We modeled daily survival rates using time-since-marking as a covariate in the nest-survival model of Program MARK. Although survival varied among and within years, we found no evidence to suggest that standard adjustment periods of 7–14 days were appropriate for our sample. If adjustment periods are used in radiotelemetry studies, those that are empirically based may be more appropriate than arbitrarily set periods.     

A novel approach for the rapid mutagenesis and directed evolution of the structural genes of west nile virus

Molecular clone technology has proven to be a powerful tool for investigating the life cycle of flaviviruses, their interactions with the host, and vaccine development. Despite the demonstrated utility of existing molecular clone strategies, the feasibility of employing these existing approaches in large-scale mutagenesis studies is limited by the technical challenges of manipulating relatively large molecular clone plasmids that can be quite unstable when propagated in bacteria. We have developed a novel strategy that provides an extremely rapid approach for the introduction of mutations into the structural genes of West Nile virus (WNV). The backbone of this technology is a truncated form of the genome into which DNA fragments harboring the structural genes are ligated and transfected directly into mammalian cells, bypassing entirely the requirement for cloning in bacteria. The transfection of cells with this system results in the rapid release of WNV that achieves a high titer (～10(7) infectious units/ml in 48 h). The suitability of this approach for large-scale mutagenesis efforts was established in two ways. First, we constructed and characterized a library of variants encoding single defined amino acid substitutions at the 92 residues of the "pr" portion of the precursor-to-membrane (prM) protein. Analysis of a subset of these variants identified a mutation that conferred resistance to neutralization by an envelope protein-specific antibody. Second, we employed this approach to accelerate the identification of mutations that allow escape from neutralizing antibodies. Populations of WNV encoding random changes in the E protein were produced in the presence of a potent monoclonal antibody, E16. Viruses resistant to neutralization were identified in a single passage. Together, we have developed a simple and rapid approach to produce infectious WNV that accelerates the process of manipulating the genome to study the structure and function of the structural genes of this important human pathogen.     

Compression or tension? The stress distribution in the proximal femur

Background  

Questions regarding the distribution of stress in the proximal human femur have never been adequately resolved. Traditionally, by considering the femur in isolation, it has been believed that the effect of body weight on the projecting neck and head places the superior aspect of the neck in tension. A minority view has proposed that this region is in compression because of muscular forces pulling the femur into the pelvis. Little has been done to study stress distributions in the proximal femur. We hypothesise that under physiological loading the majority of the proximal femur is in compression and that the internal trabecular structure functions as an arch, transferring compressive stresses to the femoral shaft.     

Fatty Acid and Chlorophyll Levels of Coastal Bermudagrass during the Day and during Maturation

Lipids were extracted from fresh, field-grown coastal bermudagrass (Cynodon dactylon (L.) Pers.) and the fatty acids determined by gas chromatography. Total fatty acid levels (dry weight basis) increased during the day and reached a single maximum at sunset in 2-week-old grass; whereas, in older grass, the fluctuations in fatty acid levels showed two maxima. The first maximum occurred 4 h after sunrise and the second maximum occurred at sunset. Total fatty acid levels, based on dry weight, decreased during the first 6 weeks of growth and changed very little after an additional 4 weeks' growth in bermudagrass leaf blades. Chlorophyll levels (dry weight basis) continuously decreased during the entire growth period (10 weeks). Chlorophyll alb ratios increased at sunset in 2- and 6-week-old grass, but this ratio did not change during the day in subsequent growth stages. The results of these experiments show that stages of maturity affected fatty acid fluctuations during the day as well as total fatty acid and chlorophyll levels in Coastal bermudagrass leaf blades. Chlorophyll alb ratios varied independently of fatty acid levels.     

A Cannibal in the National Museum: The Early Career of Franz Boas in America

Franz Boas spent several weeks at Fort Rupert, British Columbia, at the end of 1894, when he saw the Kwakiutl hamatsa ritual in situ for the first time. Soon after his return east Boas posed for a series of photographs in the U.S. National Museum for a diorama of the hamatsa dance. These photographs, now published for the first time, are a sharp reminder of Boas' constant (and sometimes forced) collaboration with the limited number of anthropological institutions in America at the end of the century, and of his personal difficulties in establishing himself professionally in America.