Isolation, characterization, and molecular cloning of a specific Mycobacterium tuberculosis antigen gene: identification of a species-specific sequence.

A rabbit polyclonal antiserum exhibiting a specific recognition pattern for Mycobacterium tuberculosis proteins was used to screen an M. tuberculosis genomic library constructed in the expression vector lambda gt11. One clone, denominated C1:10, expressed M. tuberculosis-specific determinants as part of a large fusion protein with beta-galactosidase. The gene for this protein has been sequenced, and it encodes a protein of 134 amino acids (13.8 kDa) which did not display significant homology with any of the previously reported proteins in the data bases. Hybridization studies with restriction fragments of the cloned sequence revealed that it was not present in the genomes of related mycobacteria, namely, M. bovis, M. bovis BCG, M. flavescens, M. fortuitum, M. phlei, and M. vaccae. These findings suggest that we have detected a gene, or a fragment therefrom, unique for M. tuberculosis whose nucleotide and amino acid sequences could be useful tools in the design of an improved vaccine or a diagnostic method of greater accuracy for tuberculosis.     

Molecular characterization and genetic variation of the VP7 gene of human rotaviruses isolated in Paraguay

Rotavirus is the main cause of acute diarrhea in infants and young children worldwide. In Paraguay, acute diarrhea ranks fourth among the causes of mortality in children under 4 years of age. Rotavirus was detected in 93 out of 410 patients admitted to three main hospitals in Asunción, Paraguay from August 1998 to August 2000. Samples from 64 patients were analyzed by RT-PCR for G and P typing. G4P[8] (46.9%; 30/64) was the most common strain detected, followed by G9P[8] (17.2%; 11/64) and G1P[8] (10,9%; 7/64). Since G4 was predominant during the epidemiological peaks of 1998 and 1999, four samples were sequenced and all grouped into sublineage Ic. This sublineage was reported for the first time in 1998 in Argentina, southern border of Paraguay, and it was shown to be responsible for the increased prevalence of G4 during the epidemiological season of 1998 in that country. In addition, Paraguayan G1 strains grouped in different lineages (I and II). However, G9 was predominant during the rotavirus epidemiological peak of 2000, and phylogenetic analysis of five samples grouped into a common emergent/reemergent lineage that circulates worldwide. Since vaccination could reduce the severity and the number of cases of rotavirus disease, this study suggests that a vaccine containing recently isolated variants of the most prevalent types (G1-G4) together with the emerging G9 type, might be sufficient to elicit a protective immune response against the rotavirus types circulating currently in Paraguay.     

Chlorhexidine susceptibility and Eagle effect in planktonic cells and biofilm of nosocomial isolates

European Journal of Clinical Microbiology & Infectious Diseases - The aim of this study is to evaluate the chlorhexidine gluconate (CHG) susceptibility in both planktonic cells and biofilm of...     

Prognostic factors in fetal hydronephrosis: a multivariate analysis

With the increasing use of obstetric echography fetal hydronephrosis has been reported more frequently. The purpose of this study was to identify prognostic factors associated with adverse outcome, such as renal failure and death, in fetal hydronephrosis. One hundred and forty-eight children with fetal hydronephrosis were admitted, submitted to a systematic protocol, and prospectively followed. Prognostic factors associated with fetal echography and clinical and laboratory findings on admission were studied. The median follow-up was 39 months. The analysis was conducted in two steps. In a univariate analysis, variables associated with adverse outcome were identified by the Kaplan-Meier method. The variables that were significantly associated with adverse outcome were then included in a multivariate analysis. This analysis, using the multivariate Cox’s model, was performed to identify variables that were independently associated with a worse prognosis. Only variables that remained independently associated with adverse outcome were included in the final model. After final adjustment by Cox’s multivariate model, three variables were identified as independent predictors of adverse outcome: oligohydramnios, prematurity, and glomerular filtration rate lower than 20 ml/min. Thus, in the presence of oligohydramnios, prematurity, and abnormal renal function, the medical team must plan appropriate follow-up for infants at health centers prepared to investigate and treat uropathies in newborns. Received: 24 August 1998 / Revised: 7 December 1998 / Accepted: 11 December 1998     

Differential effects of injections of anti-mu and anti-delta monoclonal antibodies on B-cell populations in adult mice: regulation of xenoreactive natural antibody-producing cells

BACKGROUND: The depletion of differential B cell and xenoreactive natural antibodies (XNA) by anti-delta and anti-mu injections was analyzed in adult mice. Sequential treatment with anti-delta and then anti-mu induces a complete depletion of B cells and XNA and represents a potential approach to induce xenograft tolerance. METHODS: Adult mice were injected with anti-mu, anti-delta, anti-delta then anti-mu, or control isotype monoclonal antibodies from day 0 to day 14. The different B-cell populations were analyzed by FACS and immunohistology. Ig production was tested by ELISA. XNA were analyzed by FACS. RESULTS: Anti-mu injections induced a depletion of IgMhigh, immature B cells, marginal zone B cells, and B1 cells and an increase of IgG-XNA production. Anti-delta injections induced mature conventional IgDhigh B-cell depletion and increased IgM-XNA production. Interestingly, sequential injections of anti-delta then anti-mu induced a depletion of immature B cells, mature B cells (MZ, B2, and B1), and XNA. CONCLUSIONS: These results demonstrate that mature B-cell depletion in adult mice can be obtained by mAb injections and depends on the surface immunoglobulin cross-linking threshold. Indeed, anti-mu mAb depleted IgMhigh B cells (MZ and B1) and anti-delta, IgDhigh B cells (B2). The differential B-cell suppression shows that conventional B cells are responsible in the IgG-XNA production and MZ and B1 cells in the IgM-XNA production. Sequential repeated injections of anti-delta then anti-mu mAb depleted all B-cell populations and suppressed the whole XNA production.     

Preliminary study of electrocautery smoke particles produced in vitro and during laparoscopic procedures

Background: The objective of this preliminary study was to describe the particles contained in cautery smoke produced during five laparoscopic procedures and verify the collection method during three laboratory experiments on ex vivo animal tissue. Methods: A cascade impactor collected the smoke according to particle size, and particle weights were calculated on an electronic microbalance. Electron microscopic analysis and energy dispersive X-ray evaluation were used to determine particle morphology and elemental composition. Results: The particles, distributed according to size on the seven rotating trays of the impactor, had diameters ranging from 0.05 to >25 m, with most being 0.1–1 m. In vitro experiments yielded more particles, especially larger (>5 m) ones, than the surgical procedures, because the cauterized specimens could be placed much closer to the cascade impactor in the laboratory environment, eliminating most obstacles to particle recovery. In the laparoscopic surgery patients, larger particles, because of their physical properties, were more likely to remain trapped in the abdomen or to drop off in the collection apparatus. Uniformly, two populations of particles were demonstrated—either large, irregular fragments (2–25 m) rich in carbon and oxygen, suggesting structural cellular components, or small homogeneous spheres (0.1–0.5 m) composed of sodium, magnesium, calcium, and potassium salts. Conclusions: This study demonstrates the presence of breathable areosols and cell-size fragments in the cautery smoke produced during laparoscopic procedures. Their exact chemical composition and potential adverse effects for patients and personnel are not known.     

Amantadine and ketamine-induced improvement of motor coordination in lurcher mutant mice

The effects of amantadine and ketamine were compared to a placebo in a coat-hanger test on lurcher mutant mice. This test measures motor coordination and is dependent on cerebellar functioning. Both drugs improved motor coordination of the cerebellar mutants in that the time taken to reach the side-bar according to a 2 paw criterion was decreased during the drugged condition in comparison to the non-drugged condition. This result indicates that NMDA receptor antagonists may improve motor coordination in animals with cerebellar disease.     

Physiodiagnostische Indikatoren bei Spannungskopfschmerzen

Practitioners often rely on physiodiagnostic indicators to corroborate the hypothesis of a muscular origin of headache. Although these indicators have been widely applied, their reliability and validity have seldom been tested empirically in headache sufferers. In a controlled double blind study, two trained raters palpated muscle tension and latent and active myogeloses of the left and right trapezius and sternocleidomastoideus muscles and measured passive head rotation flexibility. There were 28 subjects in the study, 14 suffering from tension headache and 14 matched controls. In spite of the training, interrater reliability of ratings concerning degree of muscle tension and number of latent myogeloses proved to be too low to justify subsequent use of these indicators. In keeping with the hypothesis, the various parameters of active myogeloses very clearly differentiated between the experimental groups. The hypothesis turned out not to be true for the parameters of head rotation flexibility. In subjects suffering from tension headache, no correlations could be found between the number of myogeloses of the right trapezius muscle and parameters recorded in long-term EMGs of this muscle, and no correlations could be found between the total number of myogeloses and the chronicity of headache.     

Assisted liposuction of lipomas

Assisted liposuction of lipomas is the partial or sometimes complete aspiration of the subcutaneous benign lipomas followed by the blind resection of their membrane. This procedure limits the scar and the risk of recurrence. The author purposes here this technical note.