Accuracy of Two Circulating Antigen Tests for the Diagnosis and Surveillance of Schistosoma mansoni Infection in Low-Endemicity Settings of Côte d'Ivoire.

In low-endemicity settings, current tools for the diagnosis and surveillance of schistosomiasis are often inaccurate in detecting true infection. We assessed the accuracy of an up-converting phosphor lateral flow circulating anodic antigen (UCP-LF CAA) test and a point-of-care circulating cathodic antigen (POC-CCA) urine cassette test for the diagnosis of Schistosoma mansoni. Our study was conducted in eight schools of western Côte d'Ivoire. Fifty children, aged 9-12 years, were enrolled per school. From each child, a single urine specimen and two stool specimens were collected over consecutive days for diagnostic work-up. Urine samples were subjected to UCP-LF CAA and POC-CCA tests. From each stool sample, triplicate Kato-Katz thick smears were examined. Overall, 378 children had complete data records. The prevalence of S. mansoni, as assessed by six Kato-Katz thick smears, was 4.0%. The UCP-LF CAA and POC-CCA tests revealed S. mansoni prevalence of 25.4% and 30.7%, respectively, when considering trace results as positive, and prevalence of 23.3% and 10.9% when considering trace results as negative. In the latter case, based on a composite "gold" standard, the sensitivity of UCP-LF CAA (80.7%) was considerably higher than that of POC-CCA (37.6%) and six Kato-Katz thick smears (13.8%). The negative predictive value of UCP-LF CAA, POC-CCA, and six Kato-Katz thick smears was 92.8%, 79.8%, and 74.1%, respectively. Our results confirm that UCP-LF CAA is more accurate than Kato-Katz and POC-CCA for the diagnosis of S. mansoni in low-endemicity settings.

Recommended restrictions after 131I therapy: measured doses in family members.

Absorbed doses to family members of patients treated with (131)I were measured using thermoluminescent dosimeters worn on the chest. Twenty-two patients with thyroid cancer were hospitalized for 2 d for treatment with 3,700-7,400 MBq, and 18 hyperthyroid patients were treated on an outpatient basis with 200-600 MBq. Doses were measured over periods of 15-21 d following the administration of radioiodine in 35 partners and 38 children, aged 4 mo to 25 y. These results were correlated with dose rate measurements performed with an ionization chamber, and residual thyroid uptake was assessed by scintigraphy over the same period. In the cancer group, the residual activity in thyroid remnants was less than 50 MBq in all cases at day 4 following treatment and decayed with a mean half-life of 2.2 (SD: 0.8) d. The dose measured with thermoluminescent dosimeters was lower than 0.5 mSv in all partners and children. In the hyperthyroid group, the effective half-life averaged 6.2 (SD: 1.2) d. The median of the doses measured in partners and children were 1.04 mSv (range: 0.05-5.2) and 0.13 mSv (range: 0.04-3.1), respectively. Fifteen children (88%) received less than the dose constraint of 0.5 mSv. The ICRP recommend an annual limit of 1 mSv for the members of the public. In addition, dose constraints (for example: 0.5 mSv) should be complied with whenever possible. The recommended dose limits are generally well met among family members of patients treated with 1311 for cancer. The higher doses measured in hyperthyroid patients, compared to thyroid cancer patients, relate to a higher (131)I retention by the gland and justify more extended and stringent restriction periods, based on residual thyroid activity.

Analysis, by electrospray ionization mass spectrometry, of several forms of Clostridium pasteurianum rubredoxin.

Clostridium pasteurianum rubredoxin and its recombinant counterpart purified from Escherichia coli have been analysed by electrospray ionization m.s. (e.s.i.m.s.). Whereas the N-terminal methionine of the native protein is formylated, the recombinant one has a free N-terminal methionine. E. coli cells also produce a colourless protein from the cloned gene. This protein is absent from C. pasteurianum and was shown to be zinc-substituted rubredoxin. The molecular forms of rubredoxin detected by e.s.i.m.s. depended on the experimental conditions used. Significant conversion into apo-rubredoxin occurred when the proteins were ionized at acidic pH and detected in the positive-ion mode. This conversion was quantitative in the case of Zn-rubredoxin. In contrast, when the proteins were analysed at neutral pH in the negative-ion mode, only the holoproteins, i.e. the species initially present in the solutions, were detected in the spectra. The e.s.i.m.s. experimental conditions set up here may prove useful for the analysis of other acidic metalloproteins with weakly bound metals.

Transcript mapping of the rubredoxin gene from Clostridium pasteurianum.

The transcripts of the rubredoxin gene from C. pasteurianum have been shown to have a size of approx. 230 bases by Northern blotting techniques. The transcription start has been located within 41 bases upstream of the initiator codon. The data demonstrate that the rubredoxin gene is monocistronic. Two of the three open reading frames occurring upstream of the rubredoxin gene are adjacent and appear to encode a thioredoxin (or glutaredoxin) reductase and a thioredoxin (or glutaredoxin).

Cloning, sequencing and expression in Escherichia coli of the rubredoxin gene from Clostridium pasteurianum.

A 3.9 kb BglII-HindIII DNA fragment containing the rubredoxin gene from Clostridium pasteurianum has been cloned using oligonucleotide probes designed from the protein sequence. The 2675 bp SspI-HindIII portion of this fragment has been sequenced and found to contain three open reading frames in addition to the rubredoxin gene. The putative product of one of these open reading frames is similar to various thioredoxin reductases. The rubredoxin gene translates into a sequence that differs from the previously published protein sequence in three positions, D-14, D-22 and E-48 being replaced by the corresponding amides. These changes have been confirmed by partial resequencing of the protein. Promoter-like sequences and a transcription termination signal have been found near the sequence of the rubredoxin gene, which may therefore constitute an independent transcriptional unit. Expression of C. pasteurianum rubredoxin in Escherichia coli strain JM109 has been optimized by subcloning a 476 bp SspI-SspI fragment encompassing the rubredoxin gene. Under these conditions, the latter gene was partly under the control of the lac promoter of pUC18, and the level of rubredoxin production could be increased twofold on addition of a lactose analogue, thus reaching 2-3 mg of pure protein/l of culture. Recombinant rubredoxin was produced in E. coli cells as the holoprotein, and displayed a u.v.-visible-absorption spectrum identical with that of the rubredoxin purified from C. pasteurianum. M.s. and N-terminal sequencing showed that C. pasteurianum rubredoxin expressed in E. coli differs from its native counterpart by having an unblocked N-terminal methionine.