Safety and efficacy of hepatitis B vaccine in systemic lupus erythematosus.

Hepatitis B virus (HBV) vaccination has been implicated as a potential trigger for autoimmune diseases but there are no prospective studies in lupus. We therefore assessed prospectively the safety and efficacy of immunization with recombinant DNA HBV vaccine (Euvax B; LG Life Sciences) in systemic lupus erythematosus (SLE) patients. Twenty-eight consecutive inactive SLE patients [Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) <4], age between 18 and 50 years and negative serology for HBV, were selected. Exclusion criteria were prednisone >/=20 mg/day and immunosuppressive drugs. Clinical and laboratorial assessments were obtained at study entry and one month after the three doses. In addition, a previous one year evaluation was performed using a standard electronic protocol. The mean age was 34 +/- 7.7 years and disease duration was 10.4 +/- 6.7 years. An adequate seroconversion was achieved at the end of the study (93%), although a lower frequency after the first (4%) and second dose (54%) was observed. No significant change in mean SLEDAI score was detected after each dose throughout the study (0.14 +/- 0.52 versus 0 versus 0.61 +/- 1.66 versus 0.36 +/- 1.34, P = 0.11). Reinforcing these findings, the 11% flares during vaccination was similar to the 21% observed in the previous year (P = 0.46). Furthermore, the mean prednisone dose at study entry was comparable to the end of the study (2.86 +/- 3.06 versus 4.64 +/- 8.25 mg/day, P = 0.32). In addition, the frequency of immunosuppressive therapy during the vaccination period (11%) was alike to the 14% observed in the previous year before entry (P = 0.66). Hepatitis B vaccination was safe in inactive SLE patients with an adequate vaccine response rate.

Chloroquine gestational use in systemic lupus erythematosus: assessing the risk of child ototoxicity by pure tone audiometry.

The objective of this study was to assess child chloroquine ototoxicity after its use during the gestational period in systemic lupus erythematosus (SLE). Nineteen children over four years old were evaluated: nine were exposed to chloroquine diphosphate (CDP) during gestation and 10 were born from mothers that did not take this drug before conception or anytime during pregnancy (CONTROL). Pure tone audiometry was performed in all children and high and low frequency threshold means were compared to evaluate the hearing status. All nine mothers taking CDP were exposed to this drug at least during the first trimester of pregnancy (56% during the whole gestational period) and the mean time of CDP use was 6.1 +/- 2.9 months. No significant difference was found in children of CDP and CONTROL groups regarding age (7.6 +/- 4.4 versus 12.3 +/- 7.2 years; P = 0.10, respectively) and gender (P = 0.65). Pure tone high frequency thresholds, which are the first to be affected by ototoxic drugs, presented within normal limits in children exposed or not to CDP (8.5 +/- 5.0 versus 7.4 +/- 3.6 dBHL; P = 0.55, respectively). Likewise, the mean hearing thresholds at low frequencies were also similar in both groups (11.4 +/- 4.5 versus 11.9 +/- 3.0 dBHL; P = 0.66). In conclusion, child in utero exposure to chloroquine diphosphate does not seem to induce hearing impairment as measured by pure tone audiometry, reinforcing its safe use during pregnancy of lupus patients.

Composition of cecal bile acids in ex-germfree mice inoculated with human intestinal bacteria.

Germfree (GF) mice were orally inoculated with human fecal suspension or various components of human fecal microbiota. Three weeks after the inoculation, cecal bile acid composition of these mice was examined. More than 80% of total bile acids was deconjugated in the cecal contents of ex-GF mice associated with human fecal dilutions of 10(-2) or 10(-6), or anaerobic growth from a dilution of 10(-6). In these ex-GF mice, deoxycholic acid accounted for about 20% of total bile acids. In the cecal contents of ex-GF mice associated only with clostridia, unconjugated bile acids made up less than 40% of total bile acids, about half of those in other ex-GF groups. However, the percentage of deoxycholic acid in these mice was the same as that in the other groups. These results indicate that dominant anaerobic bacterial combination is efficient for deconjugation of primary bile acids, and that clostridia in the human feces may play an important role in 7alpha-dehydroxylation of unconjugated primary bile acids in the intestine.

Transformation of bile acids and sterols by clostridia (fusiform bacteria) in Wistar rats.

The effects on bile acid and sterol transformation of clostridia (fusiform bacteria), the dominant intestinal bacteria in rodents (ca. 10(10) counts per g wet feces) were examined in Wistar rats. After inoculation of clostridia into germ-free rats and into rats previously inoculated solely with Escherichia coli, most of the endogenous bile acids were deconjugated, and cholic acid and chenodeoxycholic acid were 7alpha-dehydroxylated to deoxycholic acid and lithocholic acid, respectively. Tauro-beta-muricholic acid, another major bile acid in rats, was deconjugated, but only part of it (ca. 30%) was transformed into hyodeoxycholic acid. Cholesterol and sitosterol were also reduced to coprostanol and sitostanol, respectively. Escherichia coli transformed neither bile acids nor sterols. These data suggest that clostridia play an important role in the formation of secondary bile acids and coprostanol in rats.

[Studies on solid phase extraction of bile acids from biological matrix].

To quantify bile acids in biological samples, a solid phase extraction method was examined. This method is known as simpler procedures with less contamination compared with solvent extraction methods. Rat bile, feces and urine were used as biological samples. Since Bond Elut C18 and C8 were proved to be more suitable than the other phases (CH and SAX) so far examined using standard bile acids, Bond Elut C18 was used for biological samples. Quantification of biological sample was carried out by gas chromatography after the extracted sample was derivatized to methyl ester by treatment with trimethylsilyldiazomethane then to trifluoroacetyl ester by trifluoroacetic anhydride. On the gas chromatography, two columns (Rtx-50 and Rtx-200) were connected to the injector with Y-tube for elimination of interference. Except for a few bile acids, high recovery with less biological contamination was obtained by this solid phase extraction method.

Target epitope in the Tax protein of human T-cell leukemia virus type I recognized by class I major histocompatibility complex-restricted cytotoxic T cells.

A trans-acting regulatory gene product p40tax (Tax) of human T-cell leukemia virus type I (HTLV-I) is one of the main target antigens recognized by cytotoxic T lymphocytes (CTL) specific for HTLV-I. A CTL epitope within the Tax protein was identified in this report. HTLV-I-specific CD8+ CTL lines established from two HTLV-I carriers with HTLV-I-associated myelopathy or Sjögren syndrome were previously demonstrated to kill predominantly the target cells expressing HTLV-I Tax. The CTL from two patients showed significant levels of cytotoxicity to autologous target cells pulsed with a synthetic peptide of 24 amino acids corresponding to the amino-terminal sequences of the Tax protein. Allogeneic target cells were also sensitized for CTL by this peptide when the target cells have HLA-A2. Tax-specific cytotoxicity, detected as cytolysis of the target cells infected with vaccinia virus-HTLV-I recombinant expressing Tax protein, was almost completely inhibited by competitor cells pulsed with the synthetic peptide. This indicates that a major CTL epitope is present in this peptide. Further analysis using shorter peptides revealed that the core sequence of the CTL epitope was LLFGYPVYV at positions 11 through 19. This sequence can be aligned with the HLA-A2-specific motifs reported recently.

High-performance liquid chromatographic analysis of a new beta-lactam antibiotic, 6059-S (moxalactam).

Reversed-phase high-performance liquid chromatography was applied to the quantitative determination of a new beta-lactam antibiotic, 6059-S, ant its R- and S-epimers were resolved. The procedure was also applied to pharmaceuticals and human urine samples. Chromatographic separation was effected on a bonded hydrophobic stationary phase with two mobile phases: methanol-phosphate butter for the resolution of the epimers and methanol-tetra-n-butylammonium phosphate for the quantitation of 6059-S. For the determination of 6059-S in human urine, the latter mobile phase was used successfully without interference by the other urine components. An in vivo experiment was conducted by administering intravenously 1 g of 6059-S to seven volunteers and analysing their urine by chromatographic and microbiological assays, and a comparison of the results gave a correlation coefficient of 0.9954. One-compartment model analysis of the time-course data revealed that 6059-S was excreted in urine intact with a rate constant of 0.433h-1.