Toxoplasma gondii: Microassay to differentiate toxoplasma inhibiting factor and interleukin 2

Using a sensitive, economical, and reproducible microassay, the relationship of toxoplasma inhibiting factor to interleukin 2 has been examined. The assay developed took advantage of the observation that (1) Toxoplasma gondii tachyzoites replicated efficiently in the murine monocytic cell line, RAW 264; (2) treatment of RAW 264 cells with toxoplasma inhibiting factor prevented intracellular replication of the parasite to an extent similar to that observed with identical treatment of freshly isolated murine peritoneal exudate cells; and (3) [³H]uracil incorporation was an efficacious means to quantify replication (or inhibition of replication) of tachyzoites within the cell line. Although toxoplasma inhibiting factor and interleukin 2 were both present in the same lectin -and antigen-stimulated splenocyte supernatant fluids, results from microassays strongly suggested that the molecules were two distinct entities.     

The Effect of mei-41 on Rdna Redundancy in DROSOPHILA MELANOGASTER

The recombination and repair defective mutant, mei-41, exhibits three rather striking effects on the genetic properties and chromosomal stability of rDNA in Drosophila. First, mei-41 inhibits rDNA magnification. However, mei-9, another recombination and repair defective mutation has no similar effect. This indicates that magnification requires some, but not all, of the gene products necessary for meiotic exchange. Second, under magnifying conditions, mei-41 induces interchanges between the X rDNA and either arm of the Ybb- chromosome. These interchanges occur at high frequency and are independent of rDNA orientation. Third, in mei-41 bb+/Ybb+ males, bobbed mutants in the X, but not the Y, also arise at high frequency. Evidence suggests that these events involve the rDNA type I insertion. The recombination and repair defective properties of mei-41 together with our results regarding its unusual and specific effects involving rDNA are explained in a simple model that has general implications for chromosome structure.     

Similarity of EPR Signal IIf rise and P-680+ decay kinetics in Tris-washed chloroplast Photosystem II preparations as a function of pH

The rise time, of Signal II_f and the decay time of P-680⁺ have been measured kinetically as a function of pH by using EPR. The Photosystem II-enriched preparations which were used as samples were derived from spinach chloroplasts, and they evolved oxygen before Tris washing. The onset kinetics of Signal II_f are in agreement, within experimental error, with the fast component of the decay of an EPR signal attributable to P-680⁺. The signal II_f rise kinetics also show good agreement with published values of the pH dependence of the decay of P-680⁺ measured optically (Conjeaud, H. and Mathis, P. (1980) Biochim. Biophys. Acta 590, 353–359). These results are consistent with a model where the species Z (or D₁) responsible for Signal II_f is the immediate electron donor to P-680⁺ in tris-washed Photosystem II fragments.     

Isolation of intact polysomes from mechanically dehulled developing grain

A rapid method for obtaining large quantities of developing groats suitable for the isolation of highly intact polysomes has been developed. Developing spikelets were harvested directly from oat panicles into liquid nitrogen and then quickly passed through a dehuller. Chaff was removed by air aspiration and the resultant groats were collected directly back into liquid nitrogen. Approximately 250 g of groats could be isolated each man-hour by the above method. In comparison, only 10 g of endosperm could be collected by squeezing it out of spikelets using an endosperm mangle. Membrane-bound polysomes extracted from the immature groats were compared to those extracted from endosperm. The largest polysomes discernable as unique peaks on sucrose gradients were ten-mers and nine-mers for groats and endosperm, respectively. Polysomes isolated from both starting materials stimulated similar incorporations of [³⁵S]methionine into trichloroacetic acid-insoluble products during in vitro translations in wheat germ extract. Both polysome preparations directed the synthesis of similar high-molecular-weight proteins. Based on these criteria, polysomes from both preparations were found to be of similar intactness, although the groat starting material was much more readily obtained. The polysome classes having the maximum absorbance peak for endosperm and groat polysomes were six-mers and eight-mers, respectively.     

PHOTOSYNTHETIC QUANTUM EFFICIENCIES OF PHYTOPLANKTON FROM PERENNIALLY ICE COVERED ANTARCTIC LAKES1

A recently introduced approach far estimating the photosynthetic quantum efficiency (φ) of a freshwater or marine phytoplankton community has been applied for the first time to high latitude polar ecosystems, namely four lakes of southern Victoria Land, Antarctica. Values for φ at various depths ranged from 0.0022–0.1560 when calculated using a recommended mean extinction coefficient for phytoplankton (i.e. k?_c= 0.016). By contrast, φ ranged from 0.0037–0.0760 when calculated using an empirically estimated value for k?_c of 0.0328. If the recommended k?_c= 0.016 more closely approaches an accurate estimate, then the φ valves indicate that the phytoplankton convert light to organic carbon more efficiently than elsewhere. However, if the empirically derived k?_c= 0.0328 more closely approaches an accurate estimate, then the φ values indicate the phytoplankton trap light more efficiently than elsewhere. Although we have not resolved whether light conversion (φ) or light trapping are more efficient, the results show that the phytoplankton of these Antarctic lakes are well adapted to performing photosynthesis under extremely low light conditions.     

A simple sensitive radioreceptor assay for calcium antagonist drugs

A radioreceptor assay for calcium channel antagonist drugs described here is based on the ability of these drugs to affect ³H-nitrendipine binding to calcium channels. All the known calcium channel antagonists may be assayed in this manner. The assay can detect 10–100 nM (4 – 40 ng/ml) nimodipine, 10–100 nM (3.5 – 35 ng/ml) nifedipine, 3–30 μM (1.2 – 12 μm/ml) prenylamine, 0.1 – 1.0 μM (49 – 490 ng/ml) verapamil and 3–30 μM (1.2 – 12 μg/ml) diltiazem. These values cover the range of concentrations of calcium channel antagonists that are clinically important. As the radioreceptor assay detects active metabolites as well as the parent drugs, it should prove a useful adjunct in cardiovascular therapy. The method is more reproducible, simpler and less expensive than other methods such as high pressure liquid chromatography.     

Haemagglutinin activity in the haemolymph of individual Acrididae (grasshopper) specimens

Twenty-four individual grasshopper specimens representing four Melanoplus spp. contained similar broad-spectrum haemolymphatic haemagglutinin. The agglutinin activity showed highest titre toward human ABO and rabbit cells among nine types of erythrocytes tested. Titre values differed between individual insects but agglutination specificity toward different erythrocytes was similar. Agglutination of type-O red cells by individual grasshopper haemolymph was inhibited by 34 of 41 tested carbohydrates, carbohydrate derivatives, alcohols and chelating agents. Individual insects showed similar patterns of haemagglutination inhibition. Non-inhibitory compounds were mannose and mannose derivatives (excepting N-acetylneuraminate), several glucose derivatives, amino sugars and ethanol. The observations indicated that haemolymph from an individual grasshopper contained complex heteroagglutinin activity similar to that found in haemolymph pooled from several insects. Determination of minimal effective inhibitor concentrations confirmed the presence of heteroagglutinin activity primarily directed toward galactose and glucose and related α-linked glycosidic derivatives.     

Maternal effects and temperature-sensitive period of mutations affecting embryogenesis in Caenorhabditis elegans

We have used standard tests to investigate the nature of gene expression of a new set of temperature-sensitive mutants defining 30 emb genes (essential for embryogenesis) in the nematode Caenorhabditis elegans. The mode of gene expression as determined by progeny tests for parental effects divides the genes into four classes. For 18 genes maternal gene expression is necessary and sufficient for normal embryogenesis; for 2 genes zygotic expression is necessary and sufficient; for 7 genes either maternal or zygotic expression is sufficient; for 3 genes both maternal and zygotic expression are necessary. One mutant displayed partial paternal sufficiency. The results of temperature-shift experiments define two “execution stages,” corresponding to the limits of the temperature-sensitive period (TSP), and indicate the nature and the time of action or synthesis of the gene products. Most of the maternally expressed genes have very early execution stages indicating translation before fertilization, but some are temperature sensitive late in embryogenesis. Early execution stages for 2 zygotically necessary genes demonstrate that the zygotic genome can be active in the earliest stages of embryogenesis. All taken together, the mode of gene expression, TSP, and arrest stage (terminal phenotype) allow us to classify functionally and begin to order the genes essential for embryogenesis. The results indicate a preeminent role for maternal genes and gene products in embryogenesis, in agreement with the results of others.     

Proliferation of cells undergoing chondrogenesis in vitro

Abstract. Continuous exposure of chicken embryo limb bud mesenchyme cells undergoing chondrogenesis in vitro to [³H] thymidine ([³H]TdR) revealed that more than 90% of the cells synthesized DNA at least once during 120 h of culture. When cells were exposed to [³H]TdR for 24 h beginning at various times throughout the culture period, the percentage of cells which incorporated [³H]TdR during each period was approximately 92%. However, when the period for incorporation of radioisotope was limited to two hours, the number of cells which incorporated [³H]TdR was found to decline during chondrogenesis in vitro. This decline was coincident with the appearance of extracellular matrix material and occurred in those cells which had, and had not, expressed the cartilage phenotype.
We conclude from these studies that (1) practically all of the cells continue to proliferate while chondrogenesis is occurring in vitro, (2) there is an increase in the length of the cell cycle during chondrogenesis in vitro, and (3) withdrawal from the cell cycle is not required for differentiation of mesenchyme into cartilage.