Filamentous phage DNA cloning vectors: a noninfective mutant with a nonpolar deletion in gene III

A filamentous phage derivative, fKN16, has been constructed from the tetracyclineresistance transducing phage fd-tet by deleting a 507-base-pair (bp) segment of phage gene III. In accord with the importance of the gene III protein in infection, the infectivity of fKN16 phage is less than 10^?8 that of fd-tet phage. In contrast to most gene III amber mutants, which are polar on the downstream phage genes VI and I, fKN16 should be a nonpolar mutant since its 507-bp deletion spans an integral number of coding triplets. And indeed, two phage traits that may depend on gene VI and I function—the level of phage production and packaging into unit-length virions—appear to be normal in fKN16. High phage production coupled with very low infectivity make fKN16 suitable as a vector for DNA cloning experiments requiring a high level of biological containment. The characteristics of fKN16 as a vector were investigated in detail, using HindIII fragments of phage λ DNA as model foreign DNA. fKN16 may also be useful in studying the role of the gene III protein in the filamentous phage life cycle.     

Autoimmunity in the relatives of patients with immunodeficiency diseases.

Autoimmune disorders are reportedly more frequent than expected in immunodeficient patients and in their relatives. The hypothesis that genetic factors related to immunodeficiency may predispose to the development of autoimmunity was studied in relatives of patients with variable immunodeficiency (VID), ataxia-telangiectasia (A-T), or X-linked infantile agammaglobulinaemia (X-LA). Close relatives of patients with VID or A-T had thyroid and gastric autoantibodies significantly more frequently than did control subjects. No abnormalities were detected in unaffected relatives of X-LA patients. The increased incidence of organ-specific autoantibodies in close relatives of VID patients was confined to those families with more than one member with immunodeficiency. These data suggest that there are at least two forms of VID, one of which is associated with familial autoimmunity. It is postulated that heterozygous carriers of the A-T gene and persons with genes involved in the development of VID may exhibit T-lymphocyte dysfunction which predisposes them to autoimmunity.     

Genomic structure and evolution of the ancestral chromosome fusion site in 2q13-2q14.1 and paralogous regions on other human chromosomes

Human chromosome 2 was formed by the head-to-head fusion of two ancestral chromosomes that remained separate in other primates. Sequences that once resided near the ends of the ancestral chromosomes are now interstitially located in 2q13-2q14.1. Portions of these sequences had duplicated to other locations prior to the fusion. Here we present analyses of the genomic structure and evolutionary history of >600 kb surrounding the fusion site and closely related sequences on other human chromosomes. Sequence blocks that closely flank the inverted arrays of degenerate telomere repeats marking the fusion site are duplicated at many, primarily subtelomeric, locations. In addition, large portions of a 168-kb centromere-proximal block are duplicated at 9pter, 9p11.2, and 9q13, with 98%-99% average sequence identity. A 67-kb block on the distal side of the fusion site is highly homologous to sequences at 22qter. A third ~100-kb segment is 96% identical to a region in 2q11.2. By integrating data on the extent and similarity of these paralogous blocks, including the presence of phylogenetically informative repetitive elements, with observations of their chromosomal distribution in nonhuman primates, we infer the order of the duplications that led to their current arrangement. Several of these duplicated blocks may be associated with breakpoints of inversions that occurred during primate evolution and of recurrent chromosome rearrangements in humans.     

Hepatic clearance of Salmonella typhimurium in silica-treated mice.

Scanning electron microscopy demonstrates that crystalline silica destroys liver Kupffer cells but has no other obvious deleterious effects on the liver. Silica-treated livers still retain the ability to trap large numbers of bacteria perfused through the portal vein even though the rate of clearance is well below normal. In vivo, silica treatment decreases the rate of bacterial clearance from the blood, alters the in vivo organ distribution of cleared bacteria, and decreases the mean lethal dose of Salmonella typhimurium over 100-fold. Cumulatively, the data indicate that silica treatment enhances susceptibility to gram-negative infection, probably by destruction of macrophages.     

A Developmental Event-Related Potential Study of Picture Matching in Children, Adolescents, and Young Adults: A Replication and Extension

Event-related potentials were recorded in a developmental study of picture matching using an adaptation of Posner's (1978) letter-matching tasks. Subjects ranging in age from 6-39 were asked to decide whether two line drawings, presented sequentially, were the same or different on the basis of physical (physical identity), nominal (name identity), or categorical (category identity) criteria. The amplitude of a negativity at 400 ms (Neg400) increased as the number of dimensions on which the two line drawings differed increased. This effect held for all age groups, and was interpreted as reflecting the degree of semantic and/or physical relationship between the two pictures. However, one finding for Neg400 did suggest a qualitative difference in processing mode between the younger and older subjects. Both Neg400 and P3b latencies showed highly significant linear age trends, decreasing with increasing age. These age-related changes were interpreted as demonstrating quantitative speed of processing differences among age groups. The latencies of both Neg400 and P3b increased as the matching criteria became more complex. Moreover, P3b latency increased as the number of dimensions on which the two pictures differed increased, and this did not interact with age. Although both Neg400 and P3b showed age-related changes in scalp distribution, the fact that each was related to the experimental variables in similar fashion in all age groups suggested that they were homologous components across the age range studied. Taken as a whole, the data support continuity of information processing during these tasks across a wide age range.     

Whole organisms and purified cell walls compared as immunosorbents for the detection of IgE antibodies to Staphylococcus aureus

We have developed an immunoradiometric assay for IgE antibodies to Staphylococcus aureus (Staph IgE-Ab) which uses purified cell walls (PCW) from the Wood 46 strain of S. aureus as an immunosorbent. We compared Wood 46 PCW and whole organisms (WO) as immunosorbents for Staph IgE-Ab by performing tests on sera from patients with atopic dermatitis (AD) or the hyperimmunoglobulin E syndrome (hyper IgE syndrome). Sera with Staph IgE-Ab demonstrated dose-dependent binding to PCW and WO, but the ratio of specific to non-specific binding was much greater with PCW. Mean non-specific binding to WO was greater than to PCW, 5% versus 2%; and non-specific binding to WO varied directly with the serum concentration of IgE. Results of tests on patients' sera indicated that PCW are required in screening assays for Staph IgE-Ab to avoid false positive results caused by high levels of non-specific binding to WO.