Gilbert syndrome associated with beta-thalassemia.

The authors investigated whether the considerable variability in serum bilirubin levels (STB) found in transfusion-dependent beta-thalassemia, beta-thal intermedia, and heterozygous beta-thalassemia individuals could be related to the coexistence of Gilbert syndrome (GS). The promoter region [A(TA)nTAA] of the bilirubin UDP-glucuronosyltransferase gene (UGT1A1) was analyzed in a total of 128 beta-thalassemia individuals (108 transfusion-dependent beta-thal patients, 20 very mild beta-thal intermedia) and in 33 beta-thal heterozygotes. The control group consisted of 70 healthy children with no history of anemia. The frequency of GS genotype (TA)7/(TA)7 did not differ significantly between the groups studied. A significant difference was observed between serum bilirubin levels (STB) and GS genotypes (TA)7/(TA)7 and (TA)6/(TA)7 and also between (TA)7/(TA)7 and (TA)6/(TA)6 for all groups examined. These results confirm that the (TA)7/(TA)7 GS genotype is one of the factors accounting for the hyperbilirubinemia observed in beta-thalassemia major, intermedia, and heterozygous individuals.

Seroepidemiology of Helicobacter pylori infection in Greece.

Helicobacter pylori infection is strongly associated with chronic gastritis and peptic ulceration. As the prevalence of H. pylori infection in southern European populations is not known, a serological survey of 1069 samples from three different age groups in the Greek population was carried out with an enzyme-linked immunosorbent assay (ELISA) for antibodies to these bacteria. The antigen was an ultracentrifuged supernate of whole cell sonicates of 5 isolates of H. pylori assessed by electrophoresis and by immunoblotting with negative and positive sera. The sensitivity of the test was 97.43% and the specificity 100% for IgG antibodies; IgA and IgM antibodies to the antigen preparation were not found. Antibodies to H. pylori were detected among 39.4% of children aged 1-10 years, 67.1% of recruits (20-27 years) and 70% of blood donors (20-50 years). The prevalence of antibodies did not differ with sex in each of the age groups. The proportion of individuals with antibodies to H. pylori was higher in the younger age groups than those reported for similar age groups in western Europe.

The presence of both antivirus and antiself antibodies in sera from patients with adenovirus and influenza B.

Using the ELISA method we examined serum samples from 62 male patients aged 19-23 infected with adenovirus (serotype 7), 22 children aged 7-14 infected with influenza B (B/Norway 1/84) and 113 normal subjects aged 5-30. The infections were diagnosed serologically by complement fixation, by inhibition of hemagglutination, by ELISA and by viral culture. Moreover using enzyme-linked short-time culture assay, the production of specific antivirus antibodies and autoantibodies in vitro by spleen cells (1 x 10(6) cells/well) from normal mice and from mice immunized with adenovirus and influenza B was studied. At the same time their sera antibody titers were determined. All the serum samples were tested against the following antigens: adenovirus, influenza B, ds-DNA, actin, myosin, myoglobin, thyroglobulin, H. transferrin, H. interferon a and BSA. FV. For the further characterization of positive sera, an evaluation of specificity by competitive ELISA-test and by preparations of F(ab')2 fragments from patients' sera was also carried out. It was found that the percentage of positivity for the specific virus and other antigens was higher in the patients' samples than in the samples from the normal subjects. The specific antivirus antibody was of IgG class and their titers ranged from 1/4, 800 up to 1/19,200. Autoantibodies belonged to IgM, IgA, IgG classes and their titers ranged from 1/400 to 1/1,600. In comparison, titers of normal subjects' sera ranged from 1/150 to 1/600 and 1/150 to 1/300, respectively and both were IgG classes. Both specific virus antibodies and autoantibodies appeared at the same time. The competitive ELISA-test showed a marked inhibition (95-98%) of antivirus antibodies with the specific antigen, whereas autoantibodies were less inhibited (40-50%) by homologous antigens. The antigen-antibody reaction occurred at the Fab portion of the immunoglobulin molecule, since these fragments inhibited antibody reactivity. The same results were observed with spleen cells from immunized mice and the above-mentioned antigens when cultured in vitro.

Autoantibodies in systemic lupus erythematosus and normal subjects.

The presence of various antibodies in serum samples from patients with systemic lupus erythematosus (SLE) and from healthy subjects was investigated by ELISA, using a panel of natural antigens. Fifty-eight serum samples from 58 healthy women and 50 serum samples from 30 patients with active SLE were tested with 9 natural antigens (ds-DNA, actin, tubulin, thyroglobulin, myosin, myoglobin, human transferrin, human interferon a and BSA FV). It was found that the proportion of positive sera from healthy women at a dilution of 1/20 was almost the same as that of lupus sera at a dilution of 1/150 for nearly all antigens, while at a dilution of 1/150 the proportion of positive sera from patients with SLE was significantly higher for nearly all antigens. In lupus sera a high degree of correlation was observed between titers of anti-DNA and titers of the other antibodies. One hundred eighty-eight serum samples from 53 SLE patients, taken during exacerbation and remission of the disease were tested with ds-DNA, actin and tubulin. Antibodies (IgG) to ds-DNA actin and tubulin were found in the majority of serum samples taken during the active phase of the disease. On the other hand, very few serum samples taken during remission were found to be positive. A high degree of correlation was found between the OD of anti-actin/anti-ds-DNA (r = 0.769) and anti-tubulin/anti-ds-DNA (r = 0.829). In a competitive enzyme immunoassay for DNA, actin, tubulin, myosin and thyroglobulin, a high degree of inhibition was observed with the homologous antigens.(ABSTRACT TRUNCATED AT 250 WORDS)

Leishmaniasis in Greece I. Isolation and identification of the parasite causing human and canine visceral leishmaniasis.

Three human and 19 canine leishmanial stocks were typed according to their excreted factor serotype and the electrophoretic mobility of their MDH, GPI, G6PDH and 6PGDH and shown to be identical with regard to these characters and, thus with Leishmania donovani infantum. This verifies the opinion of earlier researchers, who suggested that the parasites which cause human and canine visceral leishmaniasis in Greece are the same organism and that dogs are the reservoir for the human infection. The complexities raised by the co-existence of human cutaneous leishmaniasis in Greece caused by L. tropica (formerly L. t. minor) are stressed. A comparison was made of the clinical symptomatology, serological diagnosis by IFA and ELISA tests and parasitological diagnosis of the human cases and canine infections.

Antiactin and antitubulin antibodies in canine visceral leishmaniasis.

Visceral leishmaniasis, a chronic and often fatal disease, is caused by the protozoan parasite Leishmania donovani. Both specific and nonspecific antibodies are produced in the course of the disease, and autoantibodies may be involved in pathogenesis. Tubulin and actin have been found to be associated with L. donovani. To learn whether antiactin and antitubulin antibodies are present in visceral leishmaniasis, we tested sera from 263 infected dogs by enzyme-linked immunosorbent assay for antibodies to the antigens L. donovani, actin, and tubulin. All samples reacted positively with L. donovani, and a high percentage reacted positively with all three antigens. Sera from 202 uninfected dogs were also tested, none reacted with L. donovani antigen, although positive reactions were observed for 8 of the samples with actin or tubulin. It was found that the antibody-antigen reaction occurred at the Fab portion of the immunoglobulin molecule. Competitive enzyme immunoassays showed that the reaction was inhibited if the positive serum was first incubated with L. donovani antigen, actin, or tubulin and then tested by enzyme-linked immunosorbent assay. These results suggest that antiactin and antitubulin antibodies are present in the sera of dogs infected with visceral leishmaniasis.

TerELISA: the ELISA test performed in Terasaki plates.

The use of Terasaki (10 microliter samples) and microtitration (100 microliter samples) plates as the solid phase in enzyme immunoassays was compared. Various antigens were used for coating the plates and antibodies present in human sera were evaluated using the same anti-human Ig antibody labelled with either beta -galactosidase, alkaline phosphatase, peroxidase or glucose oxidase. The results obtained, either by scoring with the naked eye or by absorbance reading with appropriate densitometers, showed that both plates were equally suitable and that the 4 enzymes were equally effective in detecting the same lowest quantity of antibody. A comparative evaluation using either Terasaki or microtitration plates for the quantitation of human anti-Echinococcus granulosus antibody ii 50 sera demonstrated that there was a good correlation between the two procedures (r = 0.8097). Finally, the use of glucose oxidase as the enzyme marked allowed a clear-cut distinction to be made between positive and negative samples with the naked eye alone.

Isolation of salmonella from minced meat by the use of a new procedure of enrichment.

In the examination of 526 samples of minced meat for the presence of salmonellae, a preliminary pre-enrichment in buffered peptone water was made. From this medium enrichments were made, for each sample of meat, in the usual Rappaport's medium (R25), incubated at 37 degrees C, and in Muller-Kauffmanns tetrathionate broth, incubated at 43 degrees C. Moreover, an enrichment was also made in a strongly modified Rappaport's medium, containing much less malachite green (R10), incubated at 43 degrees C. The two Rappaport's media proved to be equally effective in the isolation of salmonellae and clearly more efficient, in this respect, than the Muller-Kauffmann's broth. The new Rappaport's medium (R10), incubated at 43 degrees C, has the advantage over the usuals Rappaport's and Muller-Kauffmann's enrichment media to inhibit much more considerably the growth of competing organisms than the two other enrichment media.

[A new procedure of "Salmonella" enrichment (author's transl)]. / Noueau procédé d'enrichissement de Salmonella

We have prepared a modification of Rappaport's medium by reducing the content of themalachite green oxalate: medium R10, which is incubated at 43 degrees C instead of 37 degrees. These two modifications allow a satisfactory growth of a wide range of Salmonella and a better inhibition of competing organisms than the original medium of Rappaport incubated at 37 degrees and than Muller-Kauffmann's medium incubated at 43 degrees.

[Healthy meningococcal carriers in 1973 in Greece. Sensitivity of the isolated strains to minocycline, rifampicin and sulphadiazine (author's transl)]. / Porteurs sains de méningocoques en 1973 en Grèce et sensibilité des souches isolées à la minocycline, la rifampicin et la sulfadiazine

The frequency of healthy carriers of meningococci in Greece in 1973 has been studied by examining 1105 nasopharyngeal swabs from 731 recruits, during the four recruitment periods of this year. The frequency of healthy carriers among inductees within 24 hours from the arrival in the Camp was 38.9%. After a stay of 35 to 40 days in the training Camp the frequency of healthy carriers rose to 66.4%. Among all the soldiers examined, 24.5% were carriers of meningococci of group B, 13.2% of non-typable strains, 8.1% of autoagglutinable strains, 4.1% of meningococci of group A, 3.7% of meningococci of group C and smaller percentages of strains of groups X, Y, Z and of cross-agglutinating strains. The prevalence of carriers of meningococci of groups A and C and of autoagglutinable strains was higher among recruits who have been in the Camp for 35 to 40 days. The prevalence of carriers of the other serogroups was about the same among the inductees and the other recruits. No significant differences were found in the frequency of carriers of each serogroup among soldiers on their arrival, who were permanent residents of urban or rural areas of various parts of the country. No significant seasonal variation was noted in the frequency of carriers of each serogroup. Frequent changes of the group of meningococci harboured were noted among 374 recruits examined upon their arrival, as well as after 35 to 40 days of residence in the training Camp. Among 534 strains of meningococci examined none was resistant to either minocycline or rifampicin. Among 226 strains isolated from inductees, 44.7% were resistant to 1 mug/ml of sulphadiazine, while among 235 strains isolated from recruits after they have been in the Camp for 35 to 40 days, 57.9% were resistant to that sulphonamide.