Transient state isoelectric focusing: effects of zone load and carrier ampholyte concentration on the kinetics of defocusing and refocusing in sucrose density gradients

The kinetics of focusing, defocusing, and refocusing of l-histidyl-l-tyrosine in a sucrose density gradient have been studied utilizing a special apparatus for repetitive scanning of the isoelectric focusing column, employing uv absorption optics and a digital data acquisition system. Starting from a uniform or triangular concentration profile, the band is “focused” and a nearly linear pH gradient is formed during initial focusing. The electrical field is then abolished and free diffusion occurs. The electrical field is then reapplied (“refocusing”) and the band is allowed to sharpen, presumably reapproaching a steady state. The band width was measured quantitatively as the second moment about the mean (square of the standard deviation, σ²). In theory, measurements of σ² versus time permit the estimation of the apparent diffusion coefficient (D) and the isoelectric focusing parameter (pE). If the electrical field strength E and the pH gradient, $\text{d(}\text{pH}\text{)}\text{dx}$, were also measured, then one could calculate the slope of the pH mobility curve of the protein $\text{dM}\text{d(}\text{pH}\text{)}$ evaluated at the isoelectric point. D can be measured during the defocusing stage, and $\text{pE,}\text{D}\text{pE}\text{,}\text{or}\text{D}$ can be measured during focusing or refocusing. Several limitations and difficulties in the verification of this theory have been encountered: First, the apparent diffusion coefficient depends on zone load in approximately a linear fashion. Accordingly, it is necessary to measure D at several zone loads, and then extrapolate to zero load by linear regression techniques. Second, the ampholyte concentration has a marked effect on both D and pE. Here we have no a priori reason to extrapolate to zero ampholyte concentration. Also, at present we have no satisfactory method for measurement of E. These preliminary studies should be helpful in indicating further directions for experimental refinement and for generalization of theory.     

Why Doesn't the Creed Read "Always Be Critical"? An Examination of a Liberal Curriculum

Using ethnographic data collected in a second grade classroom over the course of a school year, this paper describes the ways in which one school's discourse of liberalism is deleteriously deployed. We view the school's discipline creed as emblematic of the school's liberal curriculum, and interrogate the effects on four African American boys in the classroom when the school enacts this creed. Despite the agency that these boys obviously had, they were unable to control the ways in which they were placed at a structural disadvantage and manipulated by a system far more powerful than they were. The results were that these four boys suffered. Not only did the intended liberal curriculum fail to be translated fully into the enacted curriculum, the liberal underpinnings of this curriculum precluded teachers and students from taking any critical stance.     

GROWTH AND DEVELOPMENT OF THE MALE GAMETOPHYTE OF GINKGO BILOBA WITHIN THE OVULE (IN VIVO)

The mature male gametophyte of Ginkgo biloba can be divided into two regions: a large saccate structure that is suspended within the fertilization chamber above the archegonia, and a pervasive, highly branched haustorial system that ramifies through the intercellular air spaces of the apex of the nucellus. This morphology appears to differ in many ways from the simpler more typical male gametophytes of most other groups of seed plants. Growth and development of the male gametophyte of Ginkgo biloba were studied using computer reconstruction techniques to generate images of the gametophyte from data derived from serial sections through the ovule. These investigations reveal that morphological development of the male gametophyte of Ginkgo biloba is divided into three distinct phases: 1) Germination, characterized by an initial brief period of diffuse growth. This phenomenon has not been described for any other seed plant male gametophyte; 2) Initiation of tip growth and the formation of a tubular body, as typifies all seed plant male gametophytes. In Ginkgo, this is accompanied by a high degree of branching, giving rise to an extensively branched haustorial system; 3) Late swelling of the proximal unbranched portion of the gametophyte resulting in formation of the saccate structure that is characteristic of the mature gametophyte. This process appears to be very similar to late development in cycad male gametophytes. Thus, despite the seemingly anomalous morphology of the mature male gametophyte of Ginkgo biloba, specific patterns of growth and development are in many ways similar to growth processes expressed by the male gametophytes of some or all major groups of seed plants.     

Parasitoid behaviour: predicting field from laboratory

Abstract.  1. Most of what is known about parasitoid behaviour comes from laboratory observations: field quantitative observations on searching parasitoids are extremely difficult to do and are rare. The basic components of Aphytis melinus 's response to California red scale ( Aonidiella aurantii ) were studied in the laboratory: encounter, rejection, drumming, probing, oviposition, and host-feeding. It was then asked whether these observations provided a reliable guide to behaviour in the field in a situation that was very different from the laboratory.
2. Field observations were carried out on bark on the trunk and interior branches of trees where live scale density is extremely high in patches, dead scale make up 90% of all scale, and could be expected to interfere with Aphytis search.
3. The laboratory observations predicted well the time taken in the field for each basic event (drumming or probing) and average times spent on a scale. Also well predicted were the distributions of times spent on drumming, probing, and total time on a scale. Rejection rates were much higher in the field. Thus, the laboratory studies predicted foraging behaviour in the field with variable success; potential explanations for observed mismatch between laboratory and field and its possible larger implications are discussed.     

The action of sodium deoxycholate on Escherichia coli

Sodium deoxycholate is used in a number of bacteriological media for the isolation and classification of gram-negative bacteria from food and the environment. Initial experiments to study the effect of deoxycholate on the growth parameters of Escherichia coli showed an increase in the lag time constant and generation time and a decrease in the growth rate constant and total cell yield of this microorganism. Cell fractionation studies indicated that sodium deoxycholate at levels used in bacteriological media interferes with the incorporation of [U-14C]glucose into the cold-trichloroacetic acid-soluble, ethanol-soluble, and trypsin-soluble cellular fractions of E. coli. Finally, sodium deoxycholate interfered with the flagellation and motility of Proteus mirabilis and E. coli. It would appear then that further improvement of the deoxycholate medium may be in order.     

Formation of Metallogenium-like structures by a manganese-oxiding fungus

Structures resembling Metallogenium spp. were observed in agar and in liquid cultures of a Mn-oxidizing basidiomycetous fungus only when Mn²⁺ was oxidized. Fungal viability was necessary for formation of the structures; Mn²⁺ concentration and the presence or absence of agar in the medium were important factors determining their morphology. Slide cultures revealed no identifiable cells in any stage of development. Fluorescent dyes that stained nucleic acids and polysaccharides in the fungal hyphae did not stain the Metallogenium-like structures. Likewise, Rhodamine 123, a fluorescent probe for membrane potential, stained fungal mitochondria, but did not stain the structures. Thin sections through the structures showed no biological membranes or other cellular features. Only the characteristic ultrastructure of biological Mn oxides were observed in serial thin sections. In agar, unfixed structures disappeared permanently during reduction of Mn oxides with hydroxylamine. Glutaraldehyde fixation stabilized these structures. Fixed structures lost most of their original phase density during reduction with hydroxylamine, but continuous microscopic observations showed that their phase density could be restored by staining with Coomassie blue. Structures that formed in liquid medium did not require stabilization with glutaraldehyde during reduction of Mn oxides. They, too, lost their original phase density during reduction with hydroxylamine; phase density could be restored by staining with cationic colloidal iron or Coomassie blue. The results suggest that the Metallogenium-like structures were formed as a result of Mn oxidation associated with exopolymers produced by the fungus.Non-standard abbreviations HEPES (N-hydroxyethylpiperazine-N-2-ethane sulfonic acid) - DAPI (4,6-diamidino-2-phenylindole) - PIPES (piperazine-N,N-bis[2-ethane sulfonic acid])     

Behavior of captive-raised rattlesnakes (Crotalus enyo) as a function of rearing conditions

Two litters of rattlesnakes (Crotalus enyo; n = 6 per litter) were raised in large cages and small plastic boxes, respectively. No significant differences in exploration of a novel environment were observed between the litters, or between either of them and a group of wild-caught C. enyo, suggesting that the common practice of rearing baby snakes in small cages has no debilitating consequences on these measures. In the absence of differences between the litters, the two samples were pooled and additional analyses revealed the existence of reliable individual variation. Experiment 2 compared the two litters and the wild-caught snakes on measures of prey-directed behavior. The two captive-reared litters did not differ, but both exhibited lower levels of strike-induced chemosensory searching than did wild-caught snakes.     

Developmental expression of 44-kDa bone phosphoprotein (osteopontin) and bone γ-carboxyglutamic acid (Gla)-containing protein (osteocalcin) in calcifying tissues of rat

New aspects of the distribution and developmental appearance of the 44-kDa bone phosphoprotein (44K BPP, also called sialoprotein I or osteopontin) and bone gamma-carboxyglutamic acid (Gla)-containing protein (BGP, also called osteocalcin) during osteogenesis and dentinogenesis were investigated with immunocytochemical techniques using monospecific, affinity-purified polyclonal antibodies. Sections from newborn rat incisors and from various bone anlagen of newborn animals and fetuses were processed for detection of 44K BPP or BGP antigenicity. In addition, histochemical reactions for detection of alkaline phosphatase or calcium salts were performed on a number of the sections. The 44K BPP appears to be synthesized and secreted by chondrocytes only in the areas of cartilage-to-bone transition; these cells could participate indirectly in the process of bone formation by providing a suitable scaffold onto which primary marrow osteoblasts attach and spread. During osteogenesis, 44K BPP is found in bone-forming cells almost concomitantly with the appearance of alkaline phosphatase and before osteoid deposition, whereas BGP is still absent during early stages of mineralization. We hypothesize that this dramatic difference between the developmental appearance of 44K BPP and BGP reflects the delayed expression of the BGP gene relative to that of 44K BPP. In long-term cultures of bone marrow from adult mice, some fibroblastic cells expressed the 44K BPP phenotype; these cells could represent early osteogenic progenitor cells. Some experiments also suggested that, as with BGP, 44K BPP or an immunologically related protein is synthesized by some odontoblasts and secreted into predentin, prior to the onset of mineralization.