Evolution equation for the Higgs field in an expanding universe.
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Biomedical subjects
Publications and source records attributed to N Weiss.
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The cuticle of the filaria Dipetalonema viteae was isolated by sonication in 1% sodiumdodecylsulphate (SDS) and in a mixture of 1% SDS and 5% B-mercaptoethanol (BME). Sonication in SDS removed all internal parts and left the cuticle intact; this was verified by light- and electronmicroscopy. Sonication and incubation of the cuticle in the mixture of SDS-BME at pH 6.8 dissolved the basal and part of the median zone of the cuticle. The epicuticle and the cortical zone remained intact. The extracts were investigated using SDS-polyacrylamide gel electrophoresis; the early extracts contained a wide variety of proteins, whereas the later steps showed a consistent pattern with a smaller number of bands. Cuticles after SDS-purification, the extract of cuticular material in SDS-BME, and the cuticles insoluble in SDS-BME were used to immunize mice; the antibodies produced were visualized by an indirect fluorescent antibody test on cryostat sections of female worms. When SDS-purified cuticles were used for immunization, antibodies directed against all organs in the filariae were found. The SDS-BME extract and the insoluble cuticular pellet stimulated the production of antibodies restricted to the cuticle of adult worms and microfilariae. The purification method opens up the possibility of further isolation and characterization of antigens from the cuticle.
Following chronic retinol (vitamin A) deprivation leading to exhaustion of liver vitamin A reserves below 50 I.U. per liver hamsters were fed diets either deficient in ("Rd":250 I.U.A./kg in experiment I, 1000 I.U.A/kg in experiment II) or enriched with retinol ("Rw":10000 I.U.A/kg in experiment I and II). After 4 weeks some of the animals (36 in experiment I, 30 in II) were infected with 150 3rd-stage larvae of D. viteae, while clean animals were kept as controls. The retinol status, the immune response (indirect fluorescent antibody test: IFAT) and parasitological parameters were examined up to 8 (experiment I) and 12 weeks (experiment II) post infection (p.i.). Rd hamsters had levelling off of weight gain or weight loss, severely deficient retinol levels in serum and liver, and high mortality. Weight gain was less in infected than in uninfected hamsters, and the capacity of infected Rw animals to restore liver retinol was significantly lower than that of uninfected Rw animals. IFAT titres were similar in Rd and in Rw animals, but microfilaraemia was significantly enhanced at 8 and 10.5 weeks p.i. in Rd hamsters. While the number of worms recovered from Rd and Rw hamsters was similar, there was a significant increase in the ratio of female to male worms in Rd hamsters. Rd hamsters in experiment I produced 3.3 times the worm mass per 100 g body-weight than Rw hamsters. Also, the average mass per female worm was significantly higher in Rd than Rw in hamsters, and this parameter was negatively correlated with the liver retinol concentration in experiment I(r = -0.89). Retinol deficiency has a marked effect on growth and fertility of D. viteae in hamsters.
Crude phosphate-buffered extracts of adult Onchocerca volvulus from savanna (Mali) and rain forest (Cameroon) areas were comparatively analysed using biochemical and immunological methods. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing revealed only minor differences between the two extracts. Out of 42 bands detectable by SDS-PAGE at least 21 were identified as glycoproteins by their affinity to concanavalin A. High resolution analysis using two dimensional gel electrophoresis (2D-G) showed marked differences in the polypeptide patterns of the two extracts. Some of the over 100 polypeptides demonstrable by Coomassie blue staining (especially at pIs between 4.3 and 5.6 and mol. wts over 64kD) were clearly different when the two extracts were compared. Antigenic differences between the two extracts could be detected by crossed immunoelectrophoresis using a rabbit anti-O. volvulus hyperimmune serum. The comparison by tandem crossed immunoelectrophoresis demonstrated clearly the existence of at least three antigenic differences, four partial identities and 13 antigenic identities between the extracts. For the identification of O. volvulus antigens serologically recognized by infected patients, we combined the 2D-G with an immunoblotting technique using a pool of highly reactive onchocerciasis sera from Mali. IgG binding antigens were then identified by incubating the blot membrane with this serum pool and with 125I-labelled protein A followed by autoradiography. IgE binding antigens were detected using a 125I-labelled anti-human IgE antiserum. Whilst the overall antigenic patterns were similar, there were, however, clear differences between the antigen preparations which gives further evidence for antigenic diversity of O. volvulus from savanna and rain forest areas.
Immunological study of individuals (aged 4 to 70 years) living in an area of Mali hyperendemic for onchocerciasis revealed an 83% prevalence of skin microfilariae (mf). Microfilariae counts from skin snips were highly age-dependent. Screening for concomitant helminth infections showed a low prevalence of hookworms and Mansonella (Dipetalonema) perstans, but neither schistosomiasis nor bancroftian filariasis. Immunological results revealed strong correlation between radioallergosorbent test (RAST) and skin test (5 and 50 ng adult O. volvulus extract), between RAST and total IgE, and between IFAT and ELISA. A negative correlation exists between mf counts and skin tests and between mf counts and RAST; the lowest median values were obtained in the group with high mf counts. Skin sensitizing antibodies were detected in most locally born children aged 4-5 years. Intradermal tests showed a high rate of sensitization to O. volvulus antigen in mf-negative children, whereas ELISA and IFAT values were significantly lower in these children than in mf-positive children. Increasing concentrations of circulating IgE antibodies were found in children aged 4-11 years by RAST, and, in individuals aged 12-19 years (age group for which mf counts sharply increase), skin testing revealed a state of anergy. In long lasting infections (adults greater than 20 years) skin reactivity was comparable to that of young children or was depressed. ELISA and IFAT achieved similar results in each age group.
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Persons who terminated membership in a prepaid group practice when compared with continuing members have less health problems as measured by bed disability and psychologic well-being, report access difficult and inconvenient, and are less likely to have established a stable relationship with a physician in the plan. Access is perceived as a problem by continuing members, as well as by a large proportion of disenrollees. Enrollees who join the plan on the basis of more direct knowledge of its actual operation are more likely to continue in the plan, but this is not accounted for by satisfaction levels. They are also more successful in establishing stable physician relationships.
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To explore the relative species specificities of the IgE and IgG antibody responses to helminth infections in man, we studied four pools of sera from patients infected with Wuchereria bancrofti, Brugia malayi, Onchocerca volvulus or Ascaris lumbricoides and ten individual sera from patients with onchocerciasis. IgE antibodies were detected by radioallergosorbent test (RAST) analysis and IgG antibodies by a Staphylococcus protein A radioimmunoassay (Staph A-RIA). Analysis of the binding curves with four different immunosorbents (prepared from antigens of B. malayi, O. volvulus, Dipetalonema viteae and A. lumbricoides) in the RAST and the binding curves with these same four antigens in the Staph A-RIA confirmed the relative species specificities for both the IgE and IgG antibody responses. Then determination of these antibody levels after specific absorption of the sera with both homologous and heterologous antigens showed that in all instances there was significantly less cross-reactivity with heterologous parasite antigens (i.e. higher species specificity) in the IgE antibody response to filarial infection than in the corresponding IgG antibody response. Such findings imply that efforts toward developing techniques for specific immunodiagnosis of filarial infections are likely to be particularly successful if focused on the IgE antibody response of exposed individuals.
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A radioallergosorbent test (RAST) was developed to detect IgE antibodies against adult Onchocerca volvulus antigens coupled to CnBr-activated Sepharose. Twenty-four out of 25 (96%) onchocerciasis sera were reactive. The lower limit of sensitivity was estimated to be at approx. 3 ng/ml IgE antibodies. Tests of sera from patients with non-filarial helminth infections showed much less cross-reactivity with RAST than with an enzyme-linked immunosorbent assay (ELISA) detecting IgG and IgM antibodies against the same antigen preparation. At a specificity comparable to that of RAST, the sensitivity of ELISA was only 61%. A heterologous antigen, prepared from female Dipetalonema viteae worms, was comparatively evaluated with O. volvulus. In RAST and ELISA, onchocerciasis sera were less reactive than against the O. volvulus antigen. Since sera from patients with non-filarial helminth infections were more reactive in RAST and almost equally reactive in ELISA using the D. viteae antigen, sensitivity was 83% for RAST and only 22% for ELISA (compared at the specificity identical to that of the O. volvulus RAST).
Specific allergic histamine release from leucocytes, radioallergosorbent tests (RAST) and indirect fluorescent antibody tests (IFAT) were applied for immunodiagnosis of schistosomiasis in patients outside endemic area. Of 10 parasitologically verified cases--all of them exhibiting a low and irregular egg output--8 were detected by histamine release, whereas 4 patient with filariasis, 3 with trichuriasis and 31 parasitologically normal controls were negative in this respect. By a combination of the histamine release test, RAST and IFAT all 10 cases were diagnosed. RAST and IFAT applied to 29 patients with active or treated schistosomiasis were positive in 14 (48%) and 20 (69%) cases, respectively.
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