Analysis of the binding of peanut agglutinin (PNA) to leukaemic cells and its relationship to T-cell differentiation.

Several leukaemias have been screened with a panel of monoclonal antibodies as well as fluoresceinated peanut lectin (FITC-PNA). Approximately 25% of T-acute lymphoblastic leukaemias (T-ALLs) were strongly positive with FITC-PNA. The staining distribution pattern did not correlate with any other monoclonal antibody used, although the phenotypes of the PNA+ T-ALLs were similar to those found on cortical thymocytes and probably reflect a more mature cellular phenotype within the T-ALL group. Some myeloid leukaemias were also PNA+ although the staining was generally weak. Several T-cell lines were examined and generally the TdT- lines showed strongest fluorescence after incubation with FITC-PNA. If these lines were induced to differentiate with 12-O-tetradecanoyl phorbol 13-acetate (TPA) they became PNA-. This was accompanied by an increase in cellular sialyl transferase activity, suggesting that one step in the differentiation process of "early' T cells is the terminal sialylation of existing oligosaccharide chains. Metabolic labelling of PNA+ T-cell lines with [35S]-methionine followed by detergent lysis and affinity chromatography on PNA-agarose showed that several bands of molecular weights 40-100,000 were bound to the column when examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis. If TPA-treated cells were examined these bands were absent.     

Mesenchymal cell activation is the rate-limiting step of granulation tissue induction.

During wound repair a 3-day lag occurs between injury and granulation tissue development. When full-thickness, 8-mm-round, excisional wounds were made in the paravertebral skin of outbred Yorkshire pigs and harvested at various times, no granulation tissue was observed before day 4. Day 4 wounds were 3% filled with granulation tissue, day 5 wounds 48% filled, and day 7 wounds 88% filled. The prerequisites for granulation tissue induction are not known but hypothetically include fibrin matrix maturation or cell activation. To examine whether matrix maturation was necessary, wounds were allowed to heal for 5 or 7 days and then aggressively curetted, resulting in the formation of fresh fibrin clots in the newly formed wound spaces. In contrast to original wounds, no lag phase was observed; wounds curetted on day 5 were 23% filled with granulation tissue 1 day later and 99% filled 3 days later, whereas wounds curetted on day 7 were 47% filled 1 day later and completely filled within 2 days. Thus, granulation tissue formation resumed promptly and independently of fibrin clot matrix maturation. This observation suggested that mesenchymal cell activation might be the rate-limiting step in granulation tissue formation. To address this hypothesis more directly, cultured porcine or human fibroblasts, grown to 80% confluence in Dulbecco's minimal essential medium plus 10% fetal calf serum, were added to new wounds. These wounds were sealed with a freshly made exogenous fibrin clot. In some wounds, platelet releasate was added to the fibrin clot. Granulation tissue did not form in day 3 wounds, which had received either fibrin alone, fibrin and platelet releasate, or fibrin and fibroblasts. In contrast, granulation tissue was observed in wounds receiving fibrin, human fibroblasts, and platelet releasate. By day 4, wounds receiving cultured human fibroblasts, fibrin, and platelet releasate were 14% filled with granulation tissue compared with less than 4% granulation tissue in control wounds. Thus, fibroblast activation is a limiting step of granulation tissue formation, and continued cell stimulation is required for accelerated development.     

Medial nucleus of the amygdala in the adult Syrian hamster: a quantitative Golgi analysis of gonadal hormonal regulation of neuronal morphology.

The medial nucleus of the amygdala (Me) processes both chemosensory and hormonal input. In the male Syrian hamster the integrity of this nucleus is essential for normal reproductive behavior. To determine if gonadal steroids modulate neuronal structure in this nucleus, the morphology of Golgi-stained neurons in the anterior and posterior regions of Me were compared in castrated and reproductively intact adult hamsters. In castrated males, neurons in the posterior, but not the anterior, region of Me undergo structural changes, as indicated by a decrease in the mean highest dendritic branch level and mean somal area compared to intact males. To further elucidate the importance of testosterone and its metabolites in maintenance of neuronal structure in the adult, seven groups of male Syrian hamsters were studied. Animals were castrated and received a blank silastic capsule or a capsule filled with either testosterone, dihydrotestosterone, or estradiol, or two capsules containing both metabolites, dihydrotestosterone and estradiol. Two groups of reproductively intact animals were included: brains from one group were processed simultaneously with the castrated and hormone-treated groups (control intact group), and the other group was processed at the beginning of the experiment. Animals were tested for mating behavior and flank glands were measured to test whether the capsules were effective in releasing the hormones into circulation. After a 12-week survival period, the brains were processed with Golgi stain and well-impregnated neurons from the posterior Me were quantitatively analyzed for somal area, highest dendritic branch, total dendritic length, and density of spines. All the measures analyzed revealed a consistent pattern of response to the different gonadal steroids. Castration resulted in a decrease in the mean somal area, the mean highest dendritic branch, and the percentage of neurons with tertiary branch segments. Dihydrotestosterone treatment also resulted in a significant decrease in somal area, mean highest dendritic branch, and percentage of neurons with tertiary dendritic branches. In addition, the total dendritic length and spine density on terminal dendrites were reduced in these brains. The remaining hormone treatment groups were not significantly different from the control group. These results suggest that in orchidectomized male Syrian hamsters, testosterone or its aromatized form, estradiol, but not dihydrotestosterone, is sufficient to maintain the normal morphology of the neurons in the posterior part of the medial nucleus of the amygdala.     

Antibody to native human immunodeficiency virus type 1 envelope glycoproteins induced by IIIB and MN recombinant gp120 vaccines. The NIAID AIDS Vaccine Evaluation Group.

The ability of antibody induced by MN and IIIB recombinant gp120 (rgp120) human immunodeficiency virus type 1 (HIV-1) vaccines the bind to oligomeric native and monomeric recombinant HIV-1 envelope glycoproteins (rgp 120) was measured in 25 uninfected, healthy adult volunteers. A major focus was to evaluate the effect of simultaneous and sequential immunization with vaccines representing different strains of HIV-1 on the ability to broaden cross-reactivity of antibodies against these and other HIV-1 strains. A flow cytometric indirect immunofluorescence assay (FIFA) to detect vaccine-induced antibody to envelope glycoprotein expressed by infected and rgp120-coated target cells was used, MN rgp120 HIV-1 vaccine given alone and coadministered with IIIB rgp120 HIV-1 vaccine elicited antibody which bound to cells infected with HIV-1MN, HIV-IIIB, HIV-1RF, and HIV-1-SF2. The presence of envelope glycoprotein-binding antibody detected by FIFA correlated to a moderate degree with functional antibody against HIV-1MN and HIV-IIIB. Priming immunization with IIIB rgp120 HIV-1 vaccine followed by booster injections of MN rgp120 HIV-1 vaccine resulted in increased cross-reactive antibody binding to these and heterologous clade B HIV-1 strains infecting cells. MN rgp120 HIV-1 vaccine given alone was better able to induce cross-reactive antibody to cells infected with heterologous HIV-1 laboratory strains than was IIIB rgp120 HIV-1 vaccine given alone. The vaccines induced binding antibody to rgp120 possessing the amino acid sequence of a clade E HIV-1 strain as measured by enzyme-linked immunosorbent assay. Levels of antibody binding to cells infected with clade B HIV-1 and cells coated with monomeric rgp120 were greater than that induced by HIV-1IIIB-based gp160 vaccines in previous studies.     

Genetic screening for Alzheimer's disease: what factors predict intentions to take a test?

The authors investigated factors that predict intention to take a genetic test for Alzheimer's disease (AD). The 449 men and women were surveyed in two groups: (a) those told that a positive result meant a 90% chance of developing AD (increased certainty) and (b) those told that a positive result meant a 50% chance of developing AD (decreased certainty). Participants completed measures of the Theory of Planned Behavior (TPB), anticipated regret, risk perception, likelihood of taking a genetic test for cancer, and AD risk factors. Just over 50% of the variance in intentions was related to TPB variables, likelihood of taking a genetic test for cancer, number of people the participants knew who had AD, experimental condition, and anticipated regret. The subjective norm was the strongest predictor of intention in the increased certainty group, whereas positive belief was the strongest predictor in the decreased certainty group.     

A nuclear mutant of Chlamydomonas that exhibits increased sensitivity to UV irradiation, reduced recombination of nuclear genes, and altered transmission of chloroplast genes

Meiotic progeny of Chlamydomonas reinhardtii normally receive chloroplast genomes only from the mt+ parent. However, exceptional zygotes, which transmit the chloroplast genomes of both parents or, more rarely, only those of the mt- parent, arise at a low frequency. Mutations at the mt(+)-linked mat-3 locus were found previously to elevate the transmission of chloroplast genomes from the mt- parent, resulting in a much higher than normal frequency of exceptional zygotes. In this paper we demonstrate that an ultraviolet-sensitive nuclear mutation mapping at the uvsE1 locus, which is unlinked to mating type, also promotes chloroplast genome transmission from the mt- parent. This mutant, which was previously shown to reduce recombination of nuclear genes in meiosis, acts synergistically with the mat-3-3 mutation to produce an extremely high frequency of exceptional zygotes. Through the use of restriction fragment length polymorphisms existing in the chloroplast genomes of C. reinhardtii and the interfertile strain C. smithii, we show that chloroplast DNA fragments from the mt- parent normally begin to disappear shortly after zygote formation. However, this process appears to be blocked totally in the absence of wild-type uvsE1 and mat-3 gene products. Our findings are consistent with the hypothesis that both gene products contribute to the mechanism responsible for uniparental inheritance of the chloroplast genome from the mt+ parent.     

Contractile activity of neonatal heart cells in culture derived from offspring of exercised pregnant rats

Summary 5-day-old neonatal offspring of exercised or non-exercised pregnant Sprague Dawley rats were used to prepare primary cultures of beating myocardial cells. The cells from the exercise group exhibited a slower beating rate for both single and aggregate cells; a larger cell size; an increased percentage of contracting cells; a greater capacity to form confluent monolayers, and a greater viability. It was concluded that exercise during the period of pregnancy produced morphological alterations in the myocardium of the progeny.     

Macrophage proteases and rheumatic diseases: Regulation of plasminogen activator by thymus-derived lymphocytes

Macrophages in culture secrete a variety of products including neutral protease activities such as plasminogen activator(s) (P.A.), collagenase and elastase. These products are not made by unstimulated macrophages, but only after induction by inflammatory stimuli, phagocytosis and lymphokines. Phagocytosis induces the prompt release of high levels of P.A. by endotoxin-primed macrophages and prolonged secretion follows uptake of non-degradable particles. Stimulation of lymphocytes results in the release of a supernatant product which enhances P.A. secretion by unstimulated mouse macrophages up to 5-fold. The production of the P.A. inducer (P.A.I.) is immunologically specific and is found in allogeneic mixed leukocyte culture (MLC) reactions, but not in syngeneic controls. The P.A. is also induced in activated macrophages from animals infected with BCG orT. cruzi and challenged with specific antigen. Production of the P.A.I. in MLC reactions depends on the presence of thymus-derived (T) lymphocytes and is closely correlated with the appearance of macrophage migration inhibition factor (MIF).The induction of macrophage P.A. and other proteases provides an important pathway for activating macrophages in delayed hypersensitivity reactions and could contribute significantly to tissue destruction in chronic inflammatory diseases in joints.     

Detection of binding antibodies to native and recombinant human immunodeficiency virus type 1 envelope glycoproteins following recombinant gp160 immunization measured by flow cytometry and enzyme immunoassays. The AIDS Vaccine Clinical Trials Network.

The ability of antibody induced by vaccination with recombinant gp160 (rgp160) to bind to native and recombinant human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins was measured. Thirty-three HIV-1-seronegative healthy adult volunteers were injected four times with 40 or 80 micrograms of an HIV-1LAV envelope glycoprotein candidate vaccine per dose. The vaccine consisted of rgp160 produced in insect tissue culture cells infected with a recombinant baculovirus which contains the gp160 gene from the HIV-1LAV strain. By using a flow cytometric indirect immunofluorescence assay (FIFA) to detect vaccine-induced antibody to native envelope glycoprotein expressed by target cells infected with HIV-1IIIB, sera from 9 of the 33 vaccinees were positive. These included sera from eight vaccinees which stained HIV-1IIIB-infected cells and sera from two vaccinees which stained target cells infected with HIV-1MN, a heterologous virus strain. None of the sera stained cells infected with the HIV-1RF strain. Envelope glycoprotein-binding antibody was more frequently detectable in an enzyme-linked immunosorbent assay (ELISA) by using rgp160 compared with that which was detectable in the FIFA with uninfected target cells which were pulsed with rgp160 antigen. Positive correlations were observed between the rgp160 FIFA and a whole-virus-lysate enzyme immunoassay, between the rgp160 FIFA and the rgp160 ELISA, and between the rgp160 ELISA and the whole-virus-lysate enzyme immunoassay. The ability of sera from some volunteers who received rgp160 vaccine to bind to HIV-1-infected cells suggests that further studies with this vaccine should be done.