Specific effects on human neutrophils of antibodies to a membrane protein constituent of neutrophil receptors for chemotactic formyl-methionyl peptides

IgG and Fab were prepared from goat antisera to MP-2, the quantitatively predominant membrane protein constituent of human neutrophil receptors for chemotactic formyl-methionyl peptides. Only 10%–25% of the f-Met-Leu-Phe combining sites of MP-2 purified from neutrophil membranes that had been solubilized in Nonidet P40 exhibited binding constants similar in magnitude to those of the receptors in intact neutrophils, while the remainder of the sites retained a mean of 2% of the affinity of native receptors. Purified MP-2 elicited IgG antibodies predominantly to framework determinants, rather than the combining site, of the f-Met-Leu-Phe receptors. IgG antibodies, but not Fab, evoked the release of significant quantities of β-glucuronidase and lysozyme from neutrophils. Saturating concentrations of Fab bound to a mean of 65,000 determinants per neutrophil, as assessed with ¹²⁵I-Fab, but failed to stimulate neutrophil chemotaxis or chemokinesis, and inhibited by 15% or less the binding of [³H]f-Met-Leu-Phe to intact neutrophils. Fab of anti-MP-2 inhibited neutrophil chemotactic responses to f-Met-Leu-Phe by up to 80%, without influencing the responses to equally chemotactic concentrations of fragments of C5 and of leukotriene B₄. Preincubation of neutrophils for 2–30 min at 37° with concentrations of f-Met-Leu-Phe which suppressed significantly the number of receptors available to [³H]f-Met-Leu-Phe, increased the number of receptors detected by ¹²⁵I-Fab of anti-MP-2, while neither fragments of C5 nor leukotriene B₄ altered the number of receptors determined by either assay. Antibodies to non-combining site determinants of chemotactic peptide receptors provide a novel immunospecific probe for studies of the regulation of neutrophil chemotaxis.     

Respiratory syncytial virus infection and virus-induced inflammation are modified by contaminants of indoor air

The airway epithelium is the first cellular component of the lung to be encountered by the particles and pathogens present in inhaled air. In addition to its role as a physical barrier, the immunological activity of the airway epithelium is an essential part of the pulmonary immune system. This means that the symptoms of lung diseases that involve immunological mechanisms are frequently exacerbated by infection of the airway epithelium with respiratory viruses. The virus-induced enhancement of immunological activity in infected epithelial cells is well characterized. However, the effects that contaminants of inhaled air have upon the infectivity and replication of respiratory viruses and the inflammation they cause, are comparatively unknown. In this study, we have shown that pre-exposure of airway epithelial cells to bacterial lipopolysaccharides or a proteolytically active house dust mite allergen, is able to, respectively, inhibit or enhance the level of cellular infection with respiratory syncytial virus and similarly alter virus-induced expression of the inflammatory chemokine interleukin-8. These results suggest that respiratory syncytial virus infection and the inflammation caused by respiratory syncytial virus may be modified by the biologically active contaminants of indoor air.     

Cloning, expression, and mapping of the Staphylococcus aureus alpha-hemolysin determinant in Escherichia coli K-12.

A fragment of Staphylococcus aureus DNA encoding the alpha-hemolysin determinant was cloned from strain Wood 46 by inserting Sau3A-generated genomic DNA fragments between the BamHI sites of the lambda replacement vector L47.1. Phages expressing alpha-hemolysin were detected by overlaying plaques formed from several thousand independent recombinant phage with erythrocytes and looking for zones of hemolysis. One phage expressing alpha-hemolysin was purified and named lambda w alpha 3. This was subsequently shown to contain a 10.2-kilobase pair insert of S. aureus DNA. A 7.6-kilobase pair HindIII fragment encoding the alpha-hemolysin was subcloned from lambda w alpha 3 into the plasmid vector pACYC184 to form the hybrid plasmid pDU1148. Escherichia coli K-12 cells harboring pDU1148 synthesized a low level of alpha-hemolysin which remained associated with the cells and was not secreted into culture supernatants. When the same strain was stabbed onto blood agar plates, no zones of hemolysis were detected after overnight growth at 37 degrees C but hemolysis developed if the plates were left at room temperature for 48 h. By introducing specific deletions or Tn5 insertions into plasmid pDU1148, the alpha-hemolysin gene was mapped to a region within a 3.3-kilobase pair EcoRI-HindIII fragment which was subcloned onto the vector plasmid pBR322. A specific enzyme-linked immunosorbent assay with peroxidase-labeled rabbit anti-alpha-hemolysin antibodies was used to measure the levels of alpha-hemolysin antigen expressed in E. coli K-12 cells harboring pDU1148 or a variety of pDU1148::Tn5 and pDU1148 deletion mutants.     

Assessment of short-term visual memory in child and adolescent psychiatric inpatients

This study provides normative data on 270 child and adolescent psychiatric inpatients using the Bender Recall technique as a measure of short-term visual memory. In addition, the authors present normative data on another immediate recall technique using the Coding section of the WISC-R. These findings are examined in light of age differences and IQ scores. Detailed instructions for administering the Coding Recall technique are presented.     

Virulence of protein A-deficient and alpha-toxin-deficient mutants of Staphylococcus aureus isolated by allele replacement.

The gene coding for protein A (spa) of Staphylococcus aureus 8325-4 has been inactivated by substituting part of the spa coding sequence for a DNA fragment specifying resistance to ethidium bromide. The in vitro-constructed spa::EtBrr substitution mutation was introduced into the S. aureus chromosome by recombinational allele replacement. Southern blot hybridization showed that the in vitro-constructed mutation was present in the chromosomal spa locus. We have previously reported the inactivation of the alpha-toxin gene (hly) by allele replacement with an in vitro-constructed hly::Emr (erythromycin resistance) mutation (M. O'Reilly, J.C.S. de Azavedo, S. Kennedy, and T.J. Foster, Microb. Pathogen. 1:125-138, 1986). A double Spa- Hly- mutant was constructed by transduction. The virulence of Spa- and Hly- mutants was tested by experimental infection of mice. When subcutaneous injections were given, Hly- mutants formed a flat, darkened lesion, whereas Hly+ strains caused a raised, cream lesion. Alpha-toxin was shown to be a major factor in forming subcutaneous lesions and in causing the death of mice injected intraperitoneally. Spa- mutants were slightly less virulent than their Spa+ counterparts, which suggests that protein A is also a virulence factor of S. aureus.     

Correlates of body image dissatisfaction among overweight women seeking weight loss

Predictors of body image dissatisfaction (BID) among obese people are poorly understood. In 79 obese women seeking weight reduction, associations with BID of self-esteem, youth teasing, adult teasing, and internalization of sociocultural appearance standards (ISAS) were studied. Analyses revealed that only self-esteem, adult teasing, and ISAS predicted BID. Results highlight the importance of adulthood self-esteem and interpersonal-cultural context--rather than childhood experiences--in predicting adulthood BID.     

Dielectric properties of low-water-content tissues

The dielectric properties of two low-water-content tissues, bone marrow and adipose tissue, were measured from 1 kHz to 1 GHz. From 1 kHz to 13 MHz, the measurements were performed using a parallel-plate capacitor method. From 10 MHz to 1 GHz, a reflection coefficient technique using an open-ended coaxial transmission line was employed. The tissue water contents ranged from 1 to almost 70% by weight. The dielectric properties correlate well with the values predicted by mixture theory. Comparison with previous results from high-water-content tissues suggests that bone marrow and adipose tissues contain less motionally altered water per unit dry volume than do the previously studied tissues with lower lipid fractions. The high degree of structural heterogeneity of these tissues was reflected in the large scatter of the data, a source of uncertainty that should be considered in practical applications of the present data.     

The effect of elevated intracranial pressure on the vibrational response of the ovine head

Although potentially fatal increases in intracranial pressure (ICP) can occur in a number of pathological conditions, there is no reliable and noninvasive procedure to detect ICP elevation and quantitatively monitor changes over time. In this experimental study, the relationships between ICP elevation and the vibrational response of the head were determined. An ovine animal model was employed in which incremental increases in ICP were elicited and directly measured through intraventricular cannulae. At each ICP increment, a vibration source elicited a flexural response of the animal's head that was measured at four locations on the skull using accelerometers. Spectral analysis of the responses showed changes in proportion to ICP change up to roughly 20 cm H₂O (15 mm Hg) above normal; a clinically significant range. Both magnitude and phase changes at frequencies between 4 and 7 kHz correlated well (γ>0.92) with ICP across the study group. These findings suggest that the vibrational response of the head can be used to monitor changes in ICP noninvasively.     

Impedance-gradient electrode reduces skin irritation induced by transthoracic defibrillation

A new type of disposable external defibrillation electrode has been developed to reduce the skin irritation commonly associated with defibrillation and synchronised cardioversion. This design employs an impedance gradient to reduce the proportion of current delivered to the electrode periphery. The temperature distribution under the new electrode was compared with that of four other types of commercially available electrodes after repeated high-energy biphasic defibrillation discharges to domestic swine. Skin temperature distributions were acquired using non-invasive thermography. Measurements of the maximum temperature rise at each electrode site, taken 3.6s after the fifth defibrillation discharge, demonstrated that the new impedance-gradient electrode produced 50–60% less skin heating than two of the three uniform-impedance electrode designs. Histological examination of erythematous sites excised 24h after defibrillation quantified the associated skin damage using a scoring protocol developed for this study. In contrast to previous studies, histological examinations demonstrated second-degree skin burns following defibrillation. The new electrode design, however, induced 44–46% less skin damage than two of the traditional uniform-impedance electrodes.