Prevalence of antibodies against heat-stable antigens from Helicobacter pylori in patients with dyspeptic symptoms and normal persons.

Heat-stable antigens from Helicobacter pylori were investigated for the detection of serum IgG, IgA and IgM antibodies against H. pylori by an ELISA technique. Antibody titers against H. pylori were measured in 167 dyspeptic patients, of whom 96 were H. pylori positive confirmed by culture or microscopy, and in 482 controls (0-98 years). Increased IgG antibody titers were found significantly more often in dyspeptic patients with active chronic gastritis than in patients with normal morphology, as well as in H. pylori-positive patients as compared to H. pylori-negative patients, independent of the endoscopic findings. The heat-stable antigens were compared with acid glycine-extracted antigens and a high degree of concordance was found in the results obtained with the two antigen preparations. The differences in the IgA antibody titers against H. pylori between H. pylori-positive and H. pylori-negative dyspeptic patients were significant and may be useful to confirm a borderline IgG result. No differences were found in IgM antibody titer between H. pylori-positive and -negative patients. The greatest age-dependent increase in IgG and IgA antibody titers was found in children, and if a lower cut-off level is used for children than for adults, as has been proposed, the proportion of people with increased antibody titers against H. pylori would be almost constant from the age of between five and 10 years until the time between 61 and 80 years. Comparison of H. pylori IgG antibodies with IgG antibodies against Campylobacter jejuni and total antibodies against cytomegalovirus (CMV) showed a greater similarity between H. pylori and C. jejuni (R = 0.51) than between H. pylori and CMV (R = 0.22). This may possibly be caused by cross-reactions between H. pylori and C. jejuni. The H. pylori heat-stabile antigen seems not to be very different from other crude H. pylori antigens like acid glycine-extracted antigens, but purification and characterization of the antigens are needed to improve antibody assays.     

Expression of the heat-modifiable major outer membrane protein of Haemophilus influenzae type b is unrelated to virulence.

The heat-modifiable major outer membrane protein (P1) of Haemophilus influenzae type b (Hib) has been shown to be both exposed on the cell surface and capable of inducing the synthesis of antibodies protective against experimental Hib disease. Chemical mutagenesis of a recombinant plasmid containing the Hib gene encoding P1 resulted in inactivation of P1 expression by this plasmid. The mutated P1 gene was transformed into Hib to obtain an isogenic mutant lacking only the ability to synthesize this surface protein. In addition, the P1 gene was inserted into a plasmid shuttle vector and used to construct a recombinant Hib strain that overexpressed the P1 protein. Lack of P1 expression did not affect the ability of Hib to grow in vitro. Neither the absence nor the overproduction of P1 affected expression of capsular polysaccharide and lipooligosaccharide by Hib. The P1-negative mutant and the P1-overexpressing strain were both as susceptible to the bactericidal activity of pooled normal human serum as was the wild-type parent strain, while the P1-negative mutant was as resistant to the bactericidal activity of normal infant rat serum as was the wild-type parent strain. The P1-negative mutant was no less virulent than was the wild-type parent strain in an animal model system, such that both the numbers of animals infected by this mutant and the mean magnitudes of the resultant bacteremias were essentially identical to those obtained with challenge by the wild-type parent strain. Similarly, overexpression of P1 did not detectably affect the virulence of Hib. These data indicate that this protective protein antigen plays no detectable role in the expression of virulence by Hib, as assessed in an animal model system.     

Interference of sugars with the binding of biotin to streptavidin and avidin

Streptavidin and avidin have found widespread use as detection reagents in immunology, biochemistry and cell biology due to their high affinity binding to biotin, but the cellular functions of these proteins are not known. We have found that various sugars interfere with the binding of streptavidin and avidin to biotin. Mannose was most effective in inhibiting the binding to biotin followed by other saccharides. The inhibitory effect is most probably due to interactions of the sugars with residues in the binding pocket of streptavidin and avidin for biotin. These results show that great caution has to be exercised in the evaluation of experiments conducted with these detection reagents in the presence of sugars.     

Hemadsorption and virulence are separable properties of Mycoplasma pneumoniae.

A selective enrichment technique was used to isolate a hemadsorption-positive revertant of a hemadsorption-negative mutant strain of Mycoplasma pneumoniae. This hemadsorption-positive revertant was shown to have simultaneously regained both the ability to attach to neuraminidase-sensitive receptors on the tracheal ring respiratory epithelium in vitro and the ability to synthesize three virulent-strain-specific proteins which were not synthesized by the hemadsorption-negative mutant. Despite the persistence of the revertant in hamster lung tissue for 9 to 12 weeks postinfection, no cytopathology was observed. Intranasal inoculation of the revertant provided limited protection against a challenge dose of virulent M. pneumoniae.     

HLA-DRB1 and -DQB1 alleles in the Brazilian population of the northeastern region of the state of São Paulo

The HLA-DRB1 and -DQB1 alleles in 161 healthy unrelated individuals, including Caucasians, Blacks and Mulattos (mixed Caucasian and Black), from the Northeastern region of the state of S?o Paulo, Brazil were analysed. The 36 different DRB1 alleles detected included not only common Caucasian alleles, but also DRB1*0411, 0807 and 1402, typical of Amerindians, and DRB1*0302, 1503, and 0804, typical of African American Blacks.     

Dissecting karyotypic patterns in renal cell carcinoma: an analysis of the accumulated cytogenetic data

Molecular cytogenetic analysis frequently shows human papillomavirus (HPV) integration near translocation breakpoints in cervical cancer cells. We have recently described a cluster of HPV18 integrations in the distal end of the common fragile site FRA8C at 8q24 in primary cervical carcinoma samples. Chromosome band 8q24 contains the MYC gene (alias c-MYC), FRA8C, and FRA8D. The MYC gene is frequently deregulated--usually by translocation or amplification--in various tumor types. In the present study, we performed a molecular cytogenetic analysis of HPV18 integration patterns and the 8q24 translocation in a primary cervical carcinoma and in HeLa cells using combined binary ratio-fluorescence in situ hybridization. Our aim was to determine how the chromosomal breaks involved in these events relate physically to the MYC gene; whether they map to the FRA8C site, the FRA8D site, or both; and how they correlate with the occurrence of DNA flexibility domains. The 8q24 translocation breakpoints mapped between stretches of integrated HPV18 sequences in the distal end of FRA8C. This region contained DNA helix flexibility clusters, several of which mapped in the vicinity of HPV integration sites and translocation breakpoints in cervical carcinomas. DNA helix flexibility clusters were also found near known MYC translocation breakpoints in Burkitt lymphomas (BL), but most BL breakpoints mapped clearly outside FRA8C. Our data revealed that FRA8C is involved in HPV integration and chromosomal translocations in cervical carcinoma; however, this fragile site is not involved in classical MYC translocations in most BLs. In the context of the familial nature of cervical cancer, FRA8C may be considered a candidate susceptibility region for cervical carcinoma.     

Quantitative measurement of cortical surface features in localization-related temporal lobe epilepsy

Differences in cortical surface features between healthy controls (n = 48) and patients with temporal lobe epilepsy (n = 46), ages 14-59, were characterized by means of advanced quantitative MRI processing techniques. Cortical surface features of interest included gyral and sulcal curvature, cortical depth, and total cortical surface area. Epilepsy patients and controls differed on measures of gyrification; the abnormalities generalized despite the focal nature of the primary epileptic process. Changes in cortical surface features were associated with increasing chronological age in both groups. Abnormalities in gyrification were associated with cognitive performance and with other morphometric measurements (e.g., surface cerebral spinal fluid). These findings are related to the literature regarding morphometric changes associated with temporal lobe epilepsy and normal aging.     

Morphology of cells and hemagglutinogens of Bordetella species: resolution of substructural units in fimbriae of Bordetella pertussis.

The morphology of cells and the hemagglutinogens isolated from cultures of Bordetella pertussis, Bordetella bronchiseptica, and Bordetella parapertussis were studied by electron microscopy with the negative-staining technique. Cells of all three species had long, thin (3 nm thick), peritrichously arranged fimbriae on the cell surface. Similar structures were found in purified hemagglutinogen preparations together with shorter fimbrial structures 3 nm thick and from 40 to 100 nm long. In one experiment, long, thin fimbriae isolated from B. pertussis were found to be arranged in a crystalline structure on the specimen grid after negative staining. Optical diffraction analysis with a filtering technique performed on micrographs of these structures revealed 12.5-nm-long substructures within individual fimbriae. Further analysis resolved each of these structures into three globules, a central globule 3.5 nm in diameter and two diametrically opposed globules 2.5 nm in diameter. Based on this substructural composition, it is suggested that subunits of the individual fimbriae are connected by fragile regions. The presence of such regions would explain the size heterogeneity of the filamentous structures observed in preparations of hemagglutinogens isolated from cultures of B. pertussis and B. bronchiseptica. The concept that the short filamentous structures present in purified preparations of hemagglutinogens originate from the surface fimbriae present on the cells is supported.     

Ring chromosome 22 and neurofibromatosis

Variable constitutional mosaicism, mos45,XY,-22/46,XY,-22,+mar/46,XY,-22,+r(22)/47,XY,-22,+r(22)+mar/ 47, XY,-22,+r(22)*2, was found in PHA-stimulated peripheral blood, in a lymphoblastoid cell line and in cultured skin fibroblasts from a mentally retarded patient with neurofibromatosis. Both the ring chromosome and the small extra marker chromosome stained positively by in situ hybridization with a chromosome 14/22-specific alphoid repeat probe. DNA dosage analysis showed constitutional loss of one copy of the arylsulfatase A gene (ARSA), consistent with its terminal location on 22q. There was no evidence of constitutional loss of D22S1 or D22S28 which flank the neurofibromatosis type 2 (NF2) locus. Analysis of two DNA samples from a skin neurofibroma indicated retainment of two copies of D22S1, whereas the results were ambiguous with respect to tumor-specific loss of one copy of D22S28. It is suggested that the development of neurofibromatosis of unclear type in two r(22) carriers might be associated with somatic mutation of the NF2 locus due to instability of the ring chromosome(s), and in analogy, that somatic mutation of either NF1 or NF2 may account for some cases of neurofibromatosis which do not meet the criteria of either NF1 or NF2. The occurrence of seminoma in the proband may be fortuitous, but could also be due to the presence of a seminoma-associated locus on chromosome 22.