[The "second life" of rifampicin]. / La deuxième vie de ta rifampicine.

Despite it was initially used and was restricted mainly for the treatment of tuberculosis, rifampicin gained in past years interest for the treatment of infections associated with prosthetic devices. Rifampicin has an excellent tissue penetration and a unique activity on bacteria in biofilms growing on the surface of prosthetic devices. The rapid emergence of resistance in bacteria constitutes the Achilles' heel and therefore rifampicin should be used with caution. This review addresses the mode of action of rifampicin and describes possible pitfalls of its use.

Effects of four-week high-fructose diet on gene expression in skeletal muscle of healthy men.

AIMS: A high-fructose diet (HFrD) may play a role in the obesity and metabolic disorders epidemic. In rodents, HFrD leads to insulin resistance and ectopic lipid deposition. In healthy humans, a four-week HFrD alters lipid homoeostasis, but does not affect insulin sensitivity or intramyocellular lipids (IMCL). The aim of this study was to investigate whether fructose may induce early molecular changes in skeletal muscle prior to the development of whole-body insulin resistance. METHODS: Muscle biopsies were taken from five healthy men who had participated in a previous four-week HFrD study, during which insulin sensitivity (hyperinsulinaemic euglycaemic clamp), and intrahepatocellular lipids and IMCL were assessed before and after HFrD. The mRNA concentrations of 16 genes involved in lipid and carbohydrate metabolism were quantified before and after HFrD by real-time quantitative PCR. RESULTS: HFrD significantly (P<0.05) increased stearoyl-CoA desaturase-1 (SCD-1) (+50%). Glucose transporter-4 (GLUT-4) decreased by 27% and acetyl-CoA carboxylase-2 decreased by 48%. A trend toward decreased peroxisomal proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha) was observed (-26%, P=0.06). All other genes showed no significant changes. CONCLUSION: HFrD led to alterations of SCD-1, GLUT-4 and PGC-1alpha, which may be early markers of insulin resistance.

Seasonal and spatial community dynamics in the meromictic Lake Cadagno.

The seasonal and spatial variations in the community structure of bacterioplankton in the meromictic alpine Lake Cadagno were examined by temporal temperature gradient gel electrophoresis (TTGE) of PCR-amplified 16S rDNA fragments. Two different amplifications were performed, one specific for the domain Bacteria (Escherichia coli positions 8-536) and another specific for the family Chromatiaceae (E. coli positions 8-1005). The latter was followed by semi-nested reamplification with the bacterial primer set, allowing comparison of the two PCR approaches by TTGE. The TTGE patterns of samples from the chemocline and the anoxic monimolimnion were essentially identical, whereas the oxic mixolimnion displayed distinctively different banding patterns. For samples from the chemocline and the monimolimnion, dominant bands in the Bacteria-specific TTGE profiles comigrated with bands obtained by the semi-nested PCR approach specific for Chromatiaceae. This observation suggested that Chromatiaceae are in high abundance in the anoxic water layer. All dominant bands were excised and sequenced. Changes in the community structure, as indicated by changes in the TTGE profiles, were observed in samples taken at different times of the year. In the chemocline, Chomatium okenii was dominant in the summer months, whereas Amoebobacter purpureus populations dominated in autumn and winter. This change was confirmed by fluorescent in situ hybridization.

Bacterial diversity and community composition in the chemocline of the meromictic alpine Lake Cadagno as revealed by 16S rDNA analysis.

Using different techniques of molecular biology we investigated the bacterial diversity of the chemocline of the meromictic Lake Cadagno. Cloning of a total community 16S rDNA PCR product and subsequent screening with a combination of amplified ribosomal DNA restriction analysis and temporal temperature gradient gel electrophoresis (TTGE) analysis revealed that 30 of 47 randomly selected clones were unique. Partial sequencing and comparative analysis indicated a high bacterial diversity dominated by the gamma-Proteobacteria (33.3%). Most of these rDNA clone sequences were not closely related to any 16S rDNA sequence in the database. In a second approach, the TTGE pattern from an environmental sample was compared with the migration of the cloned 16S rDNA fragments. Four clone types were identified on the environmental pattern by excising and sequencing comigrating bands, three of which were well represented in the library: two Chromatiaceae species and one sequence affiliated with the Desulfobulbus assemblage. Using the fluorescent in situ hybridization technique we essentially confirmed the results of the cloning experiments and the TTGE analysis.

Puerperal septic pelvic thrombophlebitis: incidence and response to heparin therapy.

OBJECTIVE: Before the availability of modern imaging studies the diagnosis of septic pelvic thrombophlebitis causing prolonged puerperal fever was difficult to confirm without surgical exploration. With the use of computed tomography infection-related pelvic phlebitis can now be confirmed, and this study was designed to determine its incidence after delivery. We also designed a randomized clinical trial to evaluate the efficacy of heparin added to antimicrobial therapy for treatment of women with septic phlebitis. STUDY DESIGN: We studied women who had pelvic infection and fever that persisted after 5 days despite adequate antimicrobial therapy with clindamycin, gentamicin, and ampicillin. After giving consent study participants underwent abdominopelvic computed tomographic imaging. Women with pelvic thrombophlebitis were randomly assigned to 1 of 2 management schemes that included continuation of antimicrobial therapy, either alone or with the addition of heparin, until the temperature was .5). The 54 women with persistent fever but without computed tomographic evidence of septic pelvic thrombophlebitis were hospitalized for a mean of 12.0 +/- 4.1 days, compared with 10.9 +/- 2.9 days for women in whom thrombosis was diagnosed (P =.14). These women were followed up for >/=3 months post partum and none showed evidence of reinfection, embolic episodes, or postphlebitic syndrome. CONCLUSIONS: The overall incidence of septic pelvic thrombophlebitis was 1:3000 deliveries. The incidence was about 1:9000 after vaginal delivery and 1:800 after cesarean section. Women given heparin in addition to antimicrobial therapy for septic thrombophlebitis did not have better outcomes than did those for whom antimicrobial therapy alone was continued. These results also do not support the common empiric practice of heparin treatment for women with persistent postpartum infection.

Molecular analysis of Methanobacterium phage psiM2.

The methanogenic archaeon Methanobacterium thermoautotrophicum Marburg is infected by the double-stranded DNA phage psiM2. The complete phage genome sequence of 26 111 bp was established. Thirty-one open reading frames (orfs), all of them organized in the same direction of transcription, were identified. On the basis of comparison of the deduced amino acid sequences to known proteins and by searching for conserved motifs, putative functions were assigned to the products of six orfs. These included three proteins involved in packaging DNA into the capsid, two putative phage structural proteins and a protein related to the Int family of site-specific recombinases. Analysis of the N-terminal amino acid sequences of three phage-encoded proteins led to the identification of two genes encoding structural proteins and of peiP, the structural gene of pseudomurein endoisopeptidase. This enzyme is involved in the lysis of host cells, and it appears to belong to a novel enzyme family. peiP was overexpressed in Escherichia coli, and its product was shown to catalyse the in vitro lysis of M. thermoautotrophicum cells. Comparison of the phage psiM2 DNA sequence with parts of the sequence of the wild-type phage psiM1 suggests that psiM2 is a deletion derivative, which formed by homologous recombination between two copies of a direct repeat.

Aerobic mineralization of 2,6-dichlorophenol by Ralstonia sp. strain RK1.

A new aerobic bacterium was isolated from the sediment of a freshwater pond close to a contaminated site at Amponville (France). It was enriched in a fixed-bed reactor fed with 2,6-dichlorophenol (2,6-DCP)as the sole carbon and energy source at pH 7.5 and room temperature. The degradation of 2,6-DCP followed Monod kinetics at low initial concentrations. At concentrations above 300 microM (50 mg.liter-1), 2,6-DCP increasingly inhibited its own degradation. The base sequence of the 16S ribosomal DNA allowed us to assign the bacterium to the genus Ralstonia (formerly Alcaligenes). The substrate spectrum of the bacterium includes toluene, benzene, chlorobenzene, phenol, and all four ortho- and para-substituted mono- and dichlorophenol isomers. Substituents other than chlorine prevented degradation. The capacity to degrade 2,6-DCP was examined in two fixed-bed reactors. The microbial population grew on and completely mineralized 2,6-DCP at 2,6-DCP concentrations up to 740 microM in continuous reactor culture supplied with H2O2 as an oxygen source. Lack of peroxide completely stopped further degradation of 2,6-DCP. Lowering the acid-neutralizing capacity of the medium to 1/10th the original capacity led to a decrease in the pH of the effluent from 7 to 6 and to a significant reduction in the degradation activity. A second fixed-bed reactor successfully removed low chlorophenol concentrations (20 to 26 microM) with hydraulic residence times of 8 to 30 min.

Imaging of puerperal septic thrombophlebitis: prospective comparison of MR imaging, CT, and sonography.

OBJECTIVE: Our objective was to compare prospectively the abilities of MR imaging, CT, and sonography to reveal puerperal septic thrombophlebitis in the pelvis. SUBJECTS AND METHODS: Seventy-six women with puerperal fever for 5 days refractory to antimicrobial therapy underwent MR imaging, CT, and sonography. We obtained unenhanced axial CT images followed by enhanced images after the administration of an oral contrast agent for which we followed a specific protocol. Axial T1- and T2-weighted spin-echo MR images with phase reconstruction and sagittal T1-weighted MR images were obtained. Real-time sonography was performed using Doppler color flow mapping and spectral waveform analysis. RESULTS: Of the 76 women, 64 completed studies with all three techniques. Ovarian vein thrombosis was diagnosed in 12 women. True-positive results were indicated when at least two of the three studies showed the presence of a clot; true-negative results were indicated when at least two of the three studies showed a lack of thrombosis. MR imaging and CT revealed both ovarian veins in all cases (64/64). Sonography revealed 33 (52%) of 64 right ovarian veins and 15 (23%) of 64 left ovarian veins. MR imaging (sensitivity, 92%; specificity, 100%) and CT (sensitivity, 100%; specificity 99%) were comparable in all but two cases. In one such case, MR imaging showed patency, CT revealed findings interpreted as thrombosis, and sonography showed flow in the partially occluded vein. In the second such case, bilateral thrombosis was seen on CT, but interpretations based on sonography and MR imaging were left-sided thrombosis only. Sonography correctly revealed six of the 12 cases of ovarian vein thrombosis. CONCLUSION: CT and MR imaging proved to be the studies of choice in the evaluation of ovarian vein thrombosis.

Prenatal diagnosis of a de novo trisomy 6q22.2-->6qter and monosomy 1pter-->1p36.3. Case report with a 2-year follow-up and a brief review of other prenatal cases of partial trisomy 6q.

We report a de novo trisom 6q22.2-->6qter and monosomy 1pter-->1p36.3 identified in amniocytes by GTG banding and FISH. While ultrasonography demonstrated malformations that did not suggest a specific chromosomal syndrome, a male infant with features consistent with trisomy 6q was born. He was followed up until 23 months, when he died after cardiac surgery. The only two other prenatal cases of trisomy 6q were compared with our patient. A literature review showed that trisomy 6q has not been reported in association with the anomalies seen by ultrasound in this case.

Molecular genetics of growth and development in Populus (Salicaceae). v. mapping quantitative trait loci affecting leaf variation.

We examined the genetic variation of leaf morphology and development in the 2-yr-old replicated plantation of an interspecific hybrid pedigree of Populus trichocarpa T. & G. and P. deltoides Marsh. via both molecular and quantitative genetic methods. Leaf traits chosen were those that show pronounced differences between the original parents, including leaf size, shape, orientation, color, structure, petiole size, and petiole cross section. Leaves were sampled from the current terminal, proleptic, and sylleptic branches. In the F2 generation, leaf traits were all significantly different among genotypes, but with significant effects due to genotype X crown-position interaction. Variation in leaf pigmentation, petiole length. And petiole length proportion appeared to be under the control of few quantitative trait loci (QTLs). More QTLs were associated with single leaf area, leaf shape, lamina angle, abaxial color, and petiole flatness, and in these traits the number of QTLs varied among crown positions. In general, the estimates of QTL numbers from Wright's biometric method were close to those derived from molecular markers. For those traits with few underlying QTLs, a single marker interval could explain from 30 to 60% of the observed phenotypic variance. For multigenic traits, certain markers contributed more substantially to the observed variation than others. Genetic cluster analysis showed developmentally related traits to be more strongly associated with each other than with unrelated traits. This finding was also supported by the QTL mapping. For example, the same chromosomal segment of linkage group L seemed to account for 20% of the phenotypic variation of all dimension-related traits, leaf size, petiole length. and midrib angle. In both traits. the P. deltoides alleles had positive effects and were dominant to the P. trichocarpa alleles. Similar relationships were also found for lamina angle. abaxial greenness, and petiole.

[Accidental insertion of a temporary pacemaker electrode in the left ventricle: radiological and electrocardiographic diagnosis]. / Akzidentelle Einlage einer passageren Schrittmacherelektrode in den linken Ventrikel: radiologische und echokardiographische Diagnose.

We report a case of unintentional puncture of the subclavian artery and insertion of a temporary pacemaker electrode in the left ventricle. Therapeutic and diagnostic strategies of this rare complication of pacemaker therapy are discussed and recommendations for safe puncture of the subclavian vein are made.

Evidence for a defective prophage on the chromosome of Methanobacterium wolfei.

Evidence shows the presence on the chromosome of Methanobacterium wolfei of a defective prophage which, by DNA-DNA hybridization, is closely related to the virulent archaeophage psi M1 of Methanobacterium thermoautotrophicum Marburg. Partial sequencing of a M. wolfei 16S rRNA gene and phylogenetic analysis indicated that this organism is more closely related to other representatives of the genus Methanobacterium than to M. thermoautotrophicum Marburg. The chromosomal region of M. wolfei encoding the putative prophage was found to be deleted for two non-contiguous segments of the phage psi M1 genome and thus encompassed only 80 to 90% of the psi M1 DNA. The prophage region was mapped to a 30 kb restriction fragment on the physical map of the M. wolfei chromosome. A randomly chosen DNA fragment was cloned from phage psi M1 DNA, as was its homologous counterpart from the chromosome of M. wolfei. The 126-bp region present in both clones exhibited 100% sequence identity.

A new placental enzyme in the metabolism of cocaine: an in vitro animal model.

OBJECTIVE: The aim of this study was to analyze placental metabolism in a genetically controlled in vitro animal model. STUDY DESIGN: Placentas from Sprague-Dawley rats were centrifuged, and microsomes were isolated. Four treatment groups were incubated with cocaine over four time periods: placental microsomes + cocaine, placental microsomes + diisopropyl fluorophosphate (an anticholinesterase) + cocaine, placental microsomes + cocaine + butyrylcholinesterase, and a blank (cocaine only). Gas chromatography was used to quantify cocaine (Limit of quantitation = 19 ng/ml) and metabolites. Gas chromatography/mass spectrometry was used to verify the identity of the metabolites. RESULTS: Butyrylcholinesterase enhanced cocaine metabolism to ecgonine methyl ester. More than 40% of cocaine was metabolized to norcocaine by rat placental when diisopropyl fluorophosphate suppressed cocaine. Norcocaine is produced by hepatic N-demethylase action on methyl-bearing nitrogen in cocaine, suggesting that placenta and liver have this capacity. Gas chromatography/mass spectrometry was essential to the identification of norcocaine, because norcocaine is frequently not identified. CONCLUSIONS: This biotransformation of cocaine to norcocaine may be a primary metabolic pathway induced in the cholinesterase-deficient placenta. This has clinical implications because norcocaine is ninefold more active physiologically than cocaine or ecgononine methylesterase.

Molecular genetics of growth and development in populus. IV. Mapping QTLs with large effects on growth, form, and phenology traits in a forest tree.

We have mapped quantitative trait loci (QTLs) for commercially important traits (stem growth and form) and an adaptive trait (spring leaf flush) in a Populus F2 generation derived from a cross between interspecific F1 hybrids (P. trichocarpa x P. deltoides). Phenotypic data were collected over a 2-year period from a replicated clonal trial containing ramets of the parental, F1, and F2 trees. Contrary to the assumptions of simple polygenic models of quantitative trait inheritance, 1-5 QTLs of large effect are responsible for a large portion of the genetic variance in each of the traits measured. For example, 44.7% of the genetic variance in stem volume after 2 years of growth is controlled by just two QTLs. QTLs governing stem basal area were found clustered with QTLs for sylleptic branch leaf area, sharing similar chromosomal position and mode of action and suggesting a pleiotropic effect of QTLs ultimately responsible for stem diameter growth.

The choice of genetic material for mechanistic studies of adaptation in forest trees.

There is considerable urgency to study the mechanisms by which forest trees track environmental change, given the prospect of possible rapid climatic changes. Environmental tracking is achieved through three basic processes: (1) expression of phenotypic plasticity at the level of the individual; and (2) evolutionary change and (3) migration, both expressed at the level of the population over generations. The current distribution and genetic architecture of a species reflect how these processes interacted in response to past climatic changes during and after the last glaciation. Part of that record is encoded in the DNA of the current generation of trees and, as a result of existing field tests, is accessible for study. These field tests include, in ascending order of genetic resolution, (1) provenance tests, (2) progeny tests, and (3) three-generation clonal tests; as well as (4) clone tests, with or without genetic structure. The suitability and limitations of these tests for mechanistic studies of environmental tracking are described, both as field installations and as sources of material for parallel in-depth studies. We conclude that they represent an important information resource, which deserves to be more effectively used by the scientific community.

Molecular genetics of growth and development in Populus. III. A genetic linkage map of a hybrid poplar composed of RFLP, STS, and RAPD markers.

We have evaluated three DNA-based marker types for linkage map construction in Populus: RFLPs detected by Southern blot hybridization, STSs detected by a combination of PCR and RFLP analysis, and RAPDs. The mapping pedigree consists of three generations, with the F1 produced by interspecific hybridization between a P. trichocarpa female and a P. deltoides male. The F2 generation was made by inbreeding to the maximum degree permitted by the dioecious mating system of Populus. The applicability of STSs and RAPDs outside the mapping pedigree has been investigated, showing that these PCR-based marker systems are well-suited to breeding designs involving interspecific hybridization. A Populus genome map (343 markers) has been constructed from a combination of all three types. The length of the Populus genome is estimated to be 2400-2800 cM.

Molecular genetics of growth and development in Populus. II. Segregation distortion due to genetic load.

Distortion of expected Mendelian segregation ratios, commonly observed in many plant taxa, has been detected in an experimental three-generation inbred pedigree of Populus founded by interspecific hybridization between P. trichocarpa and P. deltoides. An RFLP linkage map was constructed around a single locus showing severe skewing of segregation ratio against F2 trees carrying the P. trichocarpa allele in homozygous form. Several hypotheses for the mechanism of segregation distortion at this locus were tested, including directional chromosome loss, segregation of a pollen lethal allele, conflicts between genetic factors that isolate the parental species, and inbreeding depression as a result of genetic load. Breeding experiments to produce inbred and outcrossed progenies were combined with PCR-based detection of RFLPs to follow the fate of the deficient allele throughout embryo and seedling development. A recessive lethal allele, lth, inherited from the P. trichocarpa parent, was found to be tightly linked to the RFLP marker locus POP1054 and to cause embryo and seedling mortality. Heterozygotes (lth/+) appear to be phenotypically normal as embryos, seedlings, and young trees.

Quantitative genetics of growth and development in Populus. I. A three-generation comparison of tree architecture during the first 2 years of growth.

One approach to gain an insight into the genetics of tree architecture is to make use of morphologically divergent parents and study their segregating progeny in the F2 and backcross (B1) generations. This approach was chosen in the present study in which material of a three-generation pedigree growing side by side in a replicated plantation, was analyzed. The pedigree included Populus trichocarpa (T) and P. deltoides (D) parents, their F1 and F2 hybrids and their B1 hybrids to the D parent. The trees were grown in the environment of the T parent and measured for the first 2 years of growth. Nine quantitative traits were studied at the stem, branch and leaf levels of tree architecture, in which the original parents differed. Strong F1 hybrid vigor relative to the better parent (T) was expressed in growth and its components. Most quantitative traits in the F2 and B1 hybrids were intermediate between the T and D parents but displayed a wide range of variation due to segregation. The results from the analysis of variance indicated that all morphometric traits were significantly different among F2 and B1 clones, but the B1 hybrids were more sensitive to replicates than the F2. Broad-sense heritabilities (H (2)) based on clonal means ranged from moderately high to high (0.50-0.90) for the traits studied, with H (2) values varying over age. The H (2) estimates reflected greater environmental "noise" in the B1 than in the F2, presumably due to the greater proportion of maladaptive D alleles in those hybrids. In both families, sylleptic branch number and length, and leaf size on the terminal, showed strong genetic correlations with stem growth. The large divergence between the two original parents in the traits studied, combined with the high chromosome number in Populus (2n=38), makes this pedigree well suited for the estimation of the number of quantitative trait loci (QTLs) underlying quantitative variation by Wright's biometric method (1968). Variation in several traits was found to be under the control of surprisingly few major QTLs: 3-4 in 2nd-year height and diameter growth, a single QTL in stem diameter/height ratio.

The pelvis after cesarean section and vaginal delivery: normal MR findings.

OBJECTIVE: In postpartum women, pain and fever can indicate serious complications, such as phlegmon or abscess of the parametrium or bladder flap, ovarian vein thrombosis, or uterine dehiscence. The purpose of this study was to ascertain the MR appearance of the normal pelvis after uncomplicated cesarean section and vaginal delivery. SUBJECTS AND METHODS: Twenty-two patients (14 who had cesarean sections and eight who had vaginal deliveries) were imaged with a superconducting 0.35-T MR system 1-5 days after delivery. Sagittal and axial T1-weighted, proton density-weighted, and T2-weighted spin-echo images were acquired. Phase reconstruction of the axial T1-weighted sagittal images was performed to confirm flow in the ovarian veins. The ovarian veins, parametrium, bladder flap, endometrium, and uterine incision were analyzed on all images. RESULTS: The uterine incision was well seen in all 14 patients who had cesarean section. The signal characteristics of the incision suggested subacute hematoma within the myometrium, and no myometrial defects were seen. In all 13 patients with a low transverse incision, a hematoma of the bladder flap was seen. Patency was confirmed in the majority of ovarian veins (39/44). No evidence of parametrial edema or mass, which can be seen in abscess or phlegmon, was found in any of the 22 patients. The ovarian perivenous fat did not show edema, which can indicate acute ovarian vein thrombosis. Increased signal in the endometrial cavity was seen in all patients; the signal characteristics were similar to those of subacute hematoma, consistent with clinical findings. CONCLUSION: Normal findings of the postpartum pelvis on MR imaging are subacute hematoma in the uterine incision, bladder flap, and endometrial cavity. Patent ovarian veins are demonstrated in most (89%) cases.