New In-vitro Test for IgE-mediated Hypersensitivity in Man

A new simple and sensitive in-vitro method for the diagnosis of type 1 (IgE-mediated) hypersensitivity in man is described. Sliced human skin is passively sensitized by reaginic serum from allergic patients and the presence of antigen-specific IgE on the sensitized slices is detected by assay of antigen-evoked histamine release. Serum from 12 out of 14 patients with clinical respiratory allergy and positive skin tests gave significant antigen-specific histamine release. This method, which is essentially an in-vitro model of the Prausnitz-Küstner reaction, should prove of value in the diagnosis of human reaginic hypersensitivity in man.     

Cell population density and phenotypic expression of tissue culture fibroblasts from heterozygotes of Lesch-Nyhan's disease (inosinate pyrophosphorylase deficiency)

Radioautographic examination of skin fibroblasts grown in tissue culture from normal donors revealed heavy labeling of almost all cells following incubation with tritiated hypoxanthine. Cells from patients with Lesch-Nyhan's disease, lacking inosinate pyrophosphorylase, had only 10 grains or less per cell. When normal and abnormal cells were mixed prior to culture, there was a progressive increase, with culture time, in the percentage of heavily labeled cells so that by 96 hr, when the cells were confluent, over 95% of the cells were heavily labeled. Reduction of cell density by subculture produced a reversion to original values. Cultures from three obligatory heterozygotes revealed the expected mixed population of cells. This appears to be a practical approach to the identification of the heterozygote.Aided by USPHS CA08748 and GM15508, and the Health Research Council of the City of New York.     

Behavioral response of 12 ungulate species in captivity to the presence of humans

Twelve species of captive ungulates were studied to determine behavioral responses to the presence of a zookeeper within the exhibit and in front of the exhibit, with and without zoo visitors present. Significant differences in behavior occurred between species for nearly all behaviors observed. A significantly greater occurrence of vigilance and approach behavior was directed toward the zookeeper while within the exhibit relative to when the zookeeper stood in front of the exhibit. A significantly lower occurrence of eating or drinking occurred when the zookeeper was inside the exhibit. Significant differences occurred across size categories in the occurrence of approaching the zookeeper. The most frequently scored behavior was visual orientation. Statistically significant differences in the occurrence of visual orientation directed by females toward the zookeeper existed across size category, with more vigilance by species with larger body size. A statistically higher occurrence of vigilance toward the zookeeper was directed by female ungulates but not by males when the zoo was closed to the public. Although no statistical significance was found regarding the intraspecific vigilance of males, data on females revealed significant differences across species and across size categories. When data regarding vigilance toward the public were analyzed, statistically significant differences existed between species for females only. Likewise, when data regarding interspecific vigilance were examined, statistically significant differences were found across size categories for females, but not for males. The potential roles of vigilance in the wild are discussed in reference to its role in captivity.     

Ion channels in the thick ascending limb of Henle's loop.

The thick ascending limb of Henle's loop (TAL) is polarized with respect to its conductances. The luminal membrane contains a K+ conductance which is made up by the synchronous operation of 60- to 80-pS K+ channels. The basolateral membrane contains a chloride conductance. This conductance corresponds most likely to a 30- to 60-pS Cl- channel present in this membrane. Our knowledge on the properties of the K+ channels of these cells has been increased rapidly by patch clamp studies: these K+ channels are inwardly rectifying. They are highly selective for K+ over Na+, Li+ and many other cations. They do not conduct Rb+, Cs+, NH+4 or other larger cations. In fact, all these three cations as well as choline, tetraethylammonium, lidocaine, verapamil, diltiazem, quinine, quinidine and Ba2+ inhibit these K+ channels. As apparent from kinetic studies the mechanisms of inhibition are different for the various blockers. The TAL K+ channels are downregulated by increasing cytosolic Ca2+ activity. Cytosolic adenosine trisphosphate (ATP) has a similar effect. This ATP inhibition is Ca2+ dependent. The affinity to ATP is augmented by increasing Ca2+. Cytosolic alkalinity increases the open probability of these channels, and cytosolic acidification has the opposite effect. This pH dependence is very marked. A change by 0.2 pH units leads to a more than twofold change in the open-channel probability. The basolateral chloride conductance reflects the properties of an outwardly rectifying 30- to 60-pS Cl- channel. This channel behaves, in many respects, like the Cl- channels of a multitude of Cl- transporting epithelia. It is characterized by two open and two closed states. It is highly selective for Cl- as compared with larger anions, and it is inhibited reversibly by Cl- channel blockers such as 5-nitro-2-(3-phenylpropylamino)-benzoate.     

Activation of tat-defective human immunodeficiency virus by ultraviolet light

Ultraviolet light (UV) is known to cause activation of gene expression from the human immunodeficiency virus type 1 (HIV-1) promoter. To address the question of whether tat-defective HIV-1 provirus could be rescued by UV irradiation we examined its effect on HeLa cells containing integrated proviruses with tat mutations. Exposure of these cells to an optimal dose of UV resulted in the production of infectious viruses. The degree of UV activation and reversion to infectious virus appeared to depend on the nature of the original tat mutation. Two of the mutants required cocultivation with tat-expressing cells to fully generate replication competent viruses, while a third mutant required only cocultivation with H9 cells. Sequencing of cDNA from cells infected with this last mutant demonstrated that the parental mutant sequence was retained and that genotypic revertants to the wild-type as well as new mutant sequences were generated. These results suggest that tat-defective HIV-1 provirus can be activated by UV and can subsequently revert to wild-type virus. This study raises the possibility that UV exposure of immune cells in the skin plays a role in the activation of defective HIV-1 in vivo.