Hyperactivation may assist human spermatozoa to detach from intimate association with the endosalpinx

The behaviour of human spermatozoa was observed during incubation with epithelial cells isolated from the isthmic and ampullary sections of human uterine (Fallopian) tubes. During incubation, spermatozoa were observed to bind to the epithelial cells of the tube (the endosalpinx), and individual spermatozoa attached and detached at intervals. The kinematic characteristics of spermatozoa during these behaviour patterns were determined. The results showed that detached spermatozoa typically had an increased curvilinear velocity and amplitude of lateral head displacement, accompanied by a decrease in their linearity. Significantly (P < 0.01) more of the detaching spermatozoa were hyperactivated than were spermatozoa prior to attachment for both isthmic (35.3 +/- 5.5 versus 4.0 +/- 3.3%; mean +/- SEM) and ampullary (26.0 +/- 7.0 versus 2.0 +/- 1.4%) regions. Incubation with epithelial cells from either region produced no differences in any category of sperm behaviour. Furthermore, there was no significant difference between regions in the amount of time spermatozoa spent bound (33.6 +/- 12.9 and 20.6 +/- 3.0 s for isthmic and ampullary tissue respectively). These results support the hypothesis that hyperactivation may assist spermatozoa in breaking connections with epithelial cells.     

Insulin receptor overexpression in 184B5 human mammary epithelial cells induces a ligand-dependent transformed phenotype

To determine the role of the insulin receptor overexpression in breast epithelial cell transformation, the 184B5 human breast epithelial cell line was transfected with human insulin receptor cDNA. In two cell lines transfected with and overexpressing human insulin receptors (IR) (223.8 and 184.5 ng IR/10(6) cells), but not in untransfected cells, insulin binding and tyrosine kinase activity were elevated, and insulin induced a dose-dependent increase in colony formation in soft agar.     

Ca2+-dependent regulation of cGMP-stimulated phosphodiesterase from the soluble fraction of the human brain

Two forms of phosphodiesterase (F1 and F2) with different regulatory properties have been isolated from the soluble fraction of human brain cortex. F1 is the Ca(2+)-calmodulin-dependent phosphodiesterase and its activity is inhibited by calmodulin antagonists (W-7, TFP, tamoxifen) via a mechanism typical for the majority of Ca(2+)-calmodulin-dependent enzymes. F2 is activated by micromolar concentrations of cGMP (7-14-fold) and by Ca2+ ions (1.5-3-fold) in the absence of exogenous calmodulin. F2 contains a tightly bound Ca(2+)-binding component (apparently calmodulin) which does not dissociate from the enzyme in the presence of EGTA. The mechanism of calmodulin antagonists action on F2 is different from that for F1.     

Improved methodology for the electrophoretic determination of horse meat in heated foodstuffs

Using linear gradient polyacrylamide gel electrophoresis to separate constituent proteins, horse meat could be clearly distinguished from beef even after heating for 20 min at up to 120°C.     

Observations on ultrastructure of the anterior adhesive areas and other anterior glands in the monogenean, Monocotyle spiremae (Monocotylidae), from the gills of Himantura fai (Dasyatididae)

Observations with the light microscope and the scanning and transmission electron microscopes have shown that the anterior end of Monocotyle spiremae has 8 slit-like apertures on the ventrolateral margins at each side of the mouth. Gland cells located next to the pharynx produce rod-shaped secretory bodies that are conveyed in ducts that open on to the surfaces of rounded lobes inside "reservoirs" behind each ventral, slit-like aperture. Rod-shaped secretory bodies are extruded into the "reservoirs" and appear to combine and form a homogeneous secretion which may bond the ventrolateral regions of the head of the parasite to a substrate. At no stage, however, were intact rods observed outside the duct endings. Gland cells that produce an ovoid secretory body also supply the head of M. spiremae, but ducts from these open dorsal and anterior to the mouth in a region where the parasite is not known to attach. There appears to be little or no chance for the ventral rods and the dorsal ovoid secretion to mix. This is the first record of a monogenean parasite with a single type of secretion supplying the ventral surfaces of the anterior end. The rods in M. spiremae differ in some respects from the rod-shaped bodies recorded previously among gyrodactylid, dactylogyrid, capsalid and acanthocotylid monogeneans.     

Characterization of receptors with a new negative image: use in molecular docking and lead optimization

The characterization of receptor binding sites is an important aspect of molecular docking, molecular recognition, and the structure-based design process. This characterization can take several forms: the receptor surface itself can be delineated or described, the space adjacent to the surface can be chemically mapped, or a negative image of the protein binding region can be generated. In this report, we describe a new method of constructing a negative image through generation of a set of spheres. These spheres lie along the receptor surface, and their centers represent possible ligand atom positions. By the method in which they are constructed, these spheres carry a limited amount of energetic and chemical information in addition to their primary geometric information. We test the accuracy of the image by comparing sphere positions to the positions of bound ligand atoms and propose a figure of merit for such tests. Then, we use the spheres to orient ligands in enzyme active sites and show how they can be used to generate low scoring configurations more efficiently than other approaches that search orientation space. In addition, two novel applications of these spheres are described: they are used to help identify structural differences among families of enzymes and to suggest points for ligand modification in analog design.     

Cyclosporin A induces apoptosis in rat hepatocytes in culture

Salmonella typhimurium phage type (PT) or definitive type (DT) 104 is a virulent pathogen for humans and animals, particularly cattle. It has been isolated increasingly from humans and animals in the United Kingdom and several other European countries and, more recently, in the United States and Canada. Humans may acquire the infection from foods of animal origin contaminated with the infective organism. Farm families are particularly at risk of acquiring the infection by contact with infected animals or by drinking unpasteurized milk. The symptoms in cattle are watery to bloody diarrhea, a drop in milk production, pyrexia, anorexia, dehydration and depression. Infection may result in septicemic salmonellosis and, upon necropsy, a fibrinonecrotic enterocolitis may be observed. The infection occurs more commonly in the calving season than at other times. Feedlot cattle and pigs may also be affected. Prolonged carriage and shedding of the pathogen may occur. Symptoms in humans consist of diarrhea, fever, headache, nausea, abdominal pain, vomiting, and, less frequently, blood in the stool. Salmonella typhimurium DT104 strains are commonly resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline.     

Changes in the intensity of effect and spectrum of action on quantitative traits of genes in Arabidopsis under varying conditions of cultivation

Three experiments were performed in different years to study a pleiotropic effect of two marker genes A and B on quantitative traits in Arabidopsis thaliana L. (Heynh.) Experiments differed in their conditions for plant growth (light intensity and soil fertility). In experiment 1, substitution of B- by bb did not affect the duration of the sowing-flowering period, whereas substitution of A- by aa caused a 2-day delay in flowering. Experiment 2 showed that both genes affected this trait. The delay in flowering was one, two, or three days when B- was substituted by bb, A- by aa, or A-B- by aabb, respectively. Therefore, these genes were additive. Data of experiments 3 were opposite to those of experiment 1: substitution of A- by aa did not affect the trait studied, whereas substitution of B- by bb caused a 2-day elongation of the sowing-flowering period. Thus, variations in growth conditions transformed the effects of the marker genes duration of the sowing-flowering period and changed a set of genes that determined this trait. Note that effects of A and B genes on other qualitative and quantitative traits (for example, plant height) were constant in all experiments. Therefore, transformation of a gene set, which influenced the sowing-flowering period, was not related to the repression or derepression of A and B genes.