Protective Immunity and Reduced Renal Colonization Induced by Vaccines Containing Recombinant Leptospira interrogans Outer Membrane Proteins and Flagellin Adjuvant.

Leptospirosis is a global zoonotic disease caused by different Leptospira species, such as Leptospira interrogans, that colonize the renal tubules of wild and domestic animals. Thus far, attempts to develop effective leptospirosis vaccines, both for humans and animals, have failed to induce immune responses capable of conferring protection and simultaneously preventing renal colonization. In this study, we evaluated the protective immunity induced by subunit vaccines containing seven different recombinant Leptospira interrogans outer membrane proteins, including the carboxy-terminal portion of the immunoglobulinlike protein A (LigA(C)) and six novel antigens, combined with aluminum hydroxide (alum) or Salmonella flagellin (FliC) as adjuvants. Hamsters vaccinated with the different formulations elicited high antigen-specific antibody titers. Immunization with LigA(C), either with alum or flagellin, conferred protective immunity but did not prevent renal colonization. Similarly, animals immunized with LigA(C) or LigA(C) coadministered with six leptospiral proteins with alum adjuvant conferred protection but did not reduce renal colonization. In contrast, immunizing animals with the pool of seven antigens in combination with flagellin conferred protection and significantly reduced renal colonization by the pathogen. The present study emphasizes the relevance of antigen composition and added adjuvant in the efficacy of antileptospirosis subunit vaccines and shows the complex relationship between immune responses and renal colonization by the pathogen.

Does preoperative subscapularis fatty muscle infiltration really matter in anterosuperior rotator cuff tears repair outcomes? A prospective multicentric study.

INTRODUCTION: Anterosuperior (AS) rotator cuff tear describes a combined tear of the subscapularis and the supraspinatus tendons. We hypothesized that results after AS tendon repairs might be influenced by the size of the subscapularis rupture and the preoperative subscapularis muscle fatty infiltration. METHODS: A prospective multicentric study of 53 AS rotator cuff tears from five centers was performed (January 2008-January 2009). Subscapularis tendon retraction and fatty infiltration were assessed preoperatively. An ultrasonographic healing control was performed 1 year after surgery. RESULTS: Patients were on average 60 years (range, 43-75 years) and were operated on average 16 months (range, 2-72 months) after the beginning of their symptoms. The incidence of AS tears was found to be 18%. Average follow-up was 15 months (range, 12-24). The Constant-Murley (CM) score for the patients with AS ruptures improved significantly from 49 points (range, 35-51 points) preoperatively to 73 points postoperatively (range, 50-95 points)(P=0.0205). CM score gains were 26 for Lafosse group 1 ruptures and 29 for Lafosse group 2 & 3 with pre- and postoperative P values at P<0.0000001 and P<0.000001, respectively. The last follow-up CM score according to the subscapularis fatty infiltration was 70 (range, 48-95) for groups 0-1, 70 (range, 56-87) for group 2, and 56 (range, 53-88) for groups 3-4 with pre- and postoperative P values at P<0.001, P<0.001, and P<0.004, respectively. The global retear rate was 6%. DISCUSSION: Our study showed that the CM score after repairs of AS rotator cuff tears was lower in advanced subscapularis fatty infiltration. However, gains in CM scores were similar whatever the initial subscapularis fatty infiltration. The rate of tendon healing was correlated with subscapularis fatty infiltration. Subscapularis tendon rupture size was not significantly correlated with outcomes. LEVEL OF EVIDENCE: Level III.

[Allograft-prosthesis composite arthroplasty for posttraumatic floating elbow]. / Coude ballant post-traumatique traité par prothèse totale manchonnée avec une allogreffe massive.

Loss of bone stock in the elbow joint raises serious problems for reconstruction surgery. Total allograft-prosthesis composite arthroplasty is an attractive alternative to revision prosthesis and isolated osteoarticular allografts known to have many drawbacks. Use of this method is rather recent for revision procedures and tumor surgery and posttraumatic cases are rare (five reported cases). We report a case of posttraumatic floating elbow treated with this technique. At 75 months follow-up, the clinical result was excellent with the Mayo Clinic performance score at 100/100. Allograft-native bone fusion was complete and there were no complications, particularly no loosening. This composite technique is particularly well adapted for patients with major bone and joint loss. It can avoid the specific problems associated with each of the techniques used alone. The allograft reconstructs bone stock while the prosthetic component avoids the clinical expression of graft epiphyseal lysis.

[Evaluation of levofloxacin susceptibility against strains isolated from lower urinary tract infections in the community]. / Evaluation de la sensibilité à la lévofloxacine des souches isolées d'infections urinaires basses communautaires.

The aim of this study was to evaluate the in vitro activity of levofloxacin (LVX) in comparison to nalidixic acid (NAL), ofloxacin (OFX), norfloxacin (NOR), amoxicillin (AMX), cefixime (CFM), cotrimoxazole (SXT) and nitrofurantoin (FT), against 402 strains recently isolated from urine specimens in outpatient women suffering from lower urinary tract infections for which short-term treatment was not indicated. MICs were determined by the agar dilution method on Mueller-Hinton medium (Bio-Rad) according to the recommendations of the Comite de l'Antibiogramme de la Societe Francaise de Microbiologie (CA-SFM). Strains were classified as susceptible (S), intermediate (I) or resistant (R) according to the CA-SFM recommended breakpoints. Quality control was carried out using three reference strains: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853. For E. coli, the most prevalent species (345 isolates: 85.3%), susceptibilities were as follows: AMX: 60.6%, CFM: 99.1%, NAL: 94.8%, NOR: 97.4%, OFX: 97.4%, LVX: 97.4%, SXT: 84.5%, FT: 98%. This study confirms the good in vitro activity of LVX, OFX, and CFM against strains isolated from urinary tract infections in the community and particularly against E. coli, which is by far the most prevalent pathogen, 90% of strains, with more than 97% of strains being susceptible.

[In vitro activity of the pristinamycin against the isolated staphylococci in the french hospitals in 1999-2000]. / Activité in vitro de la pristinamycine vis-à-vis des staphylocoques isolés dans les hôpitaux français en 1999-2000.

One thousand six hundred and fifty clinically significant, consecutive and non redundant strains of staphylococci, including 863 Staphylococcus aureus and 787 coagulase negative staphylococci (CNS), were isolated between October 1999 and March 2000 in 35 French hospital laboratories. Susceptibilities were determined in each center by a standard diffusion method according to the recommendations of CA-SFM. Strains with vancomycin zone size diameter <17 mm were sent to the central laboratory for MIC determination of vancomycin by agar dilution, as recommended by the CA-CSFM. Frequencies of resistance to oxacillin were 38.6% for S. aureus (MRSA), 54% for the CNS, all species and 62% for S. epidermidis, respectively. The antibiotics tested showed a good activity against strains of S. aureus susceptible to oxacillin, more than 95% of strains being susceptible except for erythromycin (82.6%). Against MRSA, vancomycin and prisitinamycin had the highest rates of susceptible strains, greater than 93% for the later antibiotic. More than 92% of strains of CNS susceptible or resistant to oxacillin were sensitive to pristinamycin. Pristinamycin displayed a good activity whether the strains were constitutively or inducibly resistant to MLS(B). It comes out from this in vitro study that the rate of resistance of staphylococci to pristinamycin remains weak and stable in France. Pristinamycin is a good alternative for oral treatment of staphylococcal infections.

[Developmental lumbar stenosis in eleven French Antilles patients]. / Canal lombaire étroit constitutionnel chez des patients antillais.

PURPOSE OF THE STUDY: Developmental lumbar stenosis is a rare entity, exceptionally described in the literature. No study has been directly devoted to this condition. The purpose of the present study was to examine specific features, particularly clinical and anatomic expression, observed in a series of operated patients. MATERIAL AND METHODS: Eleven patients from the French Antilles were treated for developmental lumbar stenosis between 1996 and 2000. The Verbiest criteria were used to define canal narrowness. Signs of degeneration and presence of discal herniation were exclusion criteria. Epidemiological and clinical data were collected for the 11 patients. The degree of sagittal stenosis (fixed diameter at the bone level and mobile diameter at the discal level) was measured on computed tomography images. Transverse stenosis was determined by measuring the interpedicular and interapophyseal distances. Lateral stenosis was determined by measuring the depth of the recessus. RESULTS: These patients were young (mean age 42.4 years). Most of the clinical signs were monoradicular. Discal level stenosis predominated, generally at level L4-L5. It was generally central and lateral, sagittal and transverse. The interpedicular distance was the only diameter that remained within normal limits. Soft tissues (yellow ligaments and joint capsules) played an important role in the stenosis. DISCUSSION: The rare reports of developmental lumbar stenosis describe decompensated stenosis due to discal herniation in the adolescent. Developmental lumbar stenosis is considered to be a genetic disease and its particular high frequency in the French Antilles favors this hypothesis. The stenosis results from bony (short pedicles, hypertrophic lateral masses) and ligament (hypertrophy of the yellow ligament and joint capsules) structures. CONCLUSION: Developmental lumbar stenosis produces a global (sagittal, transverse, central, lateral) narrowing of the lumbar canal where soft tissue structures apparently play a greater role than usually thought. A prospective study examining the impact of ethnic origin is required to analyze the genetic hypothesis.

Susceptibility of Enterobacteriaceae to beta-lactam agents and fluoroquinolones: a 3-year survey in France.

OBJECTIVE: To assess trends in the susceptibility to beta-lactam agents and to fluoroquinolones of clinically relevant Enterobacteriaceae isolated over a 3-year period in 14 French hospital laboratories. METHODS: During the second quarter of 1996, 1997 and 1998, 180 consecutive non-duplicate isolates of Enterobacteriaceae were collected in each center. Sixteen beta-lactams and four quinolones were tested by the disk diffusion method. In addition, the double-disk synergy test was used to screen for the production of extended-spectrum beta-lactamase (ESBL). RESULTS: Totals of 2507, 2312 and 2506 clinical isolates were obtained in each period, respectively. The distribution of Enterobacteriaceae species according to clinical specimens and wards was similar in each study period. No significant variation in the susceptibility rates to beta-lactams and fluoroquinolones was observed, except in Klebsiella pneumoniae and Enterobacter aerogenes. The prevalence of ESBL-producing isolates decreased from 18% to 9% in the former, while it increased from 32% to 54% in the latter. At the same time, the susceptibility to ofloxacin and pefloxacin increased for K. pneumoniae (P < 0.003) and cephalosporinase-producing species (P < 0.05), except Enterobacter spp. CONCLUSION: Over the 3-year study period beta-lactams and fluoroquinolones remained highly active against Enterobacteriaceae clinical isolates, with the exception of E. aerogenes, probably as a result of the dissemination of multiresistant clones in French hospitals.

[Associated fracture of the posteromedial tubercle and the lateral process of the talus: a case report. Contribution of computed tomography]. / Fracture associée du tubercule postéro-médial et de l'apophyse latérale de l'astragale: un cas. Apport de la tomodensitométrie.

PURPOSE OF THE STUDY: Fracture of lateral process of the talus is an uncommon injury, fracture of the posteromedial tubercle of the talus is quite rare, and association of the two lesions is not reported previously. We report a case of an associated fracture of posteromedial tubercle an lateral process of the talus. PATIENT AND METHODS: The mechanism of injury was forced ankle dorsiflexion and pronation, this mechanism was responsible of an avulsion of the posteromedial tubercle by the postero talotibial ligament and a fracture of lateral process by compression between the lateral malleolus and the calcaneus. RESULTS: Conventional radiographs permitted the diagnosis of the lateral process fracture but the fracture of the posteromedial tubercle was showed only on CT scans. DISCUSSION: The fracture of the posteromedial tubercle was treated by internal fixation, through a posteromedial approach. At 12 months follow-up the patient was able to walk without pain and radiographic result was excellent (no avascular necrosis of the tubercle and no Arthrosis of subtalar joint).

[Elastic intramedullary nailing of the tibia with the Marchetti-Vicenzi nail. 43 treated cases]. / L'enclouage élastique du tibia à foyer fermé par clou de Marchetti-Vicenzi. A propos de 43 cas traités.

PURPOSE OF THE STUDY: The purpose of this study was to analyse the results of tibial intramedullary nailing using an unreamed "Universal Elastic Bundle Nail". MATERIAL AND METHODS: Forty-three intramedullary nailing of tibial shaft were done in 43 patients with recents fractures, from May 1993 and May 1996. There were 36 males and 7 females. The average age was 31.5 years (range 17-68 years). Thirty-three were injured in a traffic accident (20 motorcycles, 5 pedestrians and 8 car passengers), seven were injured in a home accident (fall) and three had a sport injury. There were 13 open fractures according to Gustilo: 5 grade I, 7 grade II and one grade III B. Eight fractures involved the proximal metaphyseal part of the tibia, 16 the distal metaphyseal part and 14 the tibial shaft; in five cases there were segmental fractures. According to AO classification there were: 10 fractures type A, 24 fractures type B and 9 fractures type C (5 segmental fractures). In 5 cases there were associated femoral fractures: three ipsilaterals and two controlaterals. All were treated in the same time: four by UEBN device and one by AO's nail. All the patients with type B and C fractures were positioned on a Maquet table with a boot traction or transcalcaneal pin traction (in the distal fractures). The nail was introduced after closed reduction through a vertical transpatellar tendon incision, without reaming procedure. RESULTS: Forty one fractures healed after an average time of 96 days (60-120). In 11 open fractures bone union occurred after 98 days (85-120). The distal fractures healed after a mean time of 86 days (60-120), proximal fractures in 123 days and mid shaft fractures in 98 days. In type A fractures bone union occurred after an average time of 68 days, while bone union occurred after a mean time of 100 days in type B and C fractures. Two patients with an open proximal type B fracture, had a delayed union: both healed after proximal screws removal. Two fractures healed with a valgus angulaton 5 degrees and 10 degrees. No infection, no loss of reduction and no bundle migration has been noted. DISCUSSION: The Marchetti-Vicenzi's nail (UEBN) permitted a stable fixation in tibial fractures. The use of this unreamed nailing coupled with an automatic distal locking in the metaphyseal cancellous bone, reduced operative time and shortened X Ray's radiation exposure. At the follow-up fracture healing occurred in 41 cases 95.3 p. 100 at four months. Two delayed union occurred after four months, the two cases were open fractures grade II. All the two cases healed after secondary procedure without any loss of function. Malunion occurred in two patients (in only one case there was a major valgus angulation 10 degrees), the two cases were related to technical error. We had no cases of infection or leg shortening or bundle migration in the ankle joint. CONCLUSION: We believe that Universal Elastic Bundle Nail allows a stable and safety fixation in open or closed tibial fractures without pseudarthrosis and without infection (in our series).

[Reconstruction of a complex defect of the upper third of the humerus by pedicle transplant of the lateral edge of the scapula. A case report]. / Reconstruction d'une perte de substance complexe du tiers supérieur de l'humérus par transplant pediculé du bord latéral de l'omoplate. A propos de un cas.

Vascularized bone transfer of the lateral border of the scapula is exceptionally used in orthopaedic surgery. The authors report a case of pedicle transfer of the lateral border of the scapula designed to reconstruct a nine centimetre bone defect of the upper third of the humerus following a gunshot wound. A complementary conventional bone graft was performed one month later and consolidation was achieved at the fourth month.

New susceptibility locus for rheumatoid arthritis suggested by a genome-wide linkage study.

Rheumatoid arthritis (RA), the most common autoimmune disease, is associated in families with other autoimmune diseases, including insulin-dependent diabetes mellitus (IDDM). Its genetic component has been suggested by familial aggregation (lambdas = 5), twin studies, and segregation analysis. HLA, which is the only susceptibility locus known, has been estimated to account for one-third of this component. The aim of this paper was to identify new RA loci. A genome scan was performed with 114 European Caucasian RA sib pairs from 97 nuclear families. Linkage was significant only for HLA (P < 2.5.10(-5)) and nominal for 19 markers in 14 other regions (P < 0.05). Four of the loci implicated in IDDM potentially overlap with these regions: the putative IDDM6, IDDM9, IDDM13, and DXS998 loci. The first two of these candidate regions, defined in the RA genome scan by the markers D18S68-D18S61-D18S469 (18q22-23) and D3S1267 (3q13), respectively, were studied in 194 additional RA sib pairs from 164 nuclear families. Support for linkage to chromosome 3 only was extended significantly (P = 0.002). The analysis of all 261 families provided a linkage evidence of P = 0. 001 and suggested an interaction between this putative RA locus and HLA. This locus could account for 16% of the genetic component of RA. Candidate genes include those coding for CD80 and CD86, molecules involved in antigen-specific T cell recognition. In conclusion, this first genome scan in RA Caucasian families revealed 14 candidate regions, one of which was supported further by the study of a second set of families.

Genome search in celiac disease.

Celiac disease (CD), a malabsorption disorder of the small intestine, results from ingestion of gluten. The HLA risk factors involved in CD are well known but do not explain the entire genetic susceptibility. To determine the localization of other genetic risk factors, a systematic screening of the genome has been undertaken. The typing information of 281 markers on 110 affected sib pairs and their parents was used to test linkage. Systematic linkage analysis was first performed on 39 pairs in which both sibs had a symptomatic form of CD. Replication of the regions of interest was then carried out on 71 pairs in which one sib had a symptomatic form and the other a silent form of CD. In addition to the HLA loci, our study suggests that a risk factor in 5qter is involved in both forms of CD (symptomatic and silent). Furthermore, a factor on 11qter possibly differentiates the two forms. In contrast, none of the regions recently published was confirmed by the present screening.

European Gene Mapping Project (EUROGEM): breakpoint panels for human chromosomes based on the CEPH reference families. Centre d'Etude du Polymorphisme Humain.

Meiotic breakpoint panels for human chromosomes 2, 3, 4, 5, 6, 7, 8, 9, 10, 13, 14, 15, 17, 18, 20 and X were constructed from genotypes from the CEPH reference families. Each recombinant chromosome included has a breakpoint well-supported with reference to defined quantitative criteria. The panels were constructed at both a low-resolution, useful for a first-pass localization, and high-resolution, for a more precise placement. The availability of such panels will reduce the number of genotyping experiments necessary to order new polymorphisms with respect to existing genetic markers. This paper shows only a representative sample of the breakpoints detected. The complete data are available on the World Wide Web (URL http:/(/)www.icnet.uk/axp/hgr/eurogem++ +/HTML/data.html) or by anonymous ftp (ftp.gene.ucl.ac.uk in/pub/eurogem/maps/breakpoints).

A radiation hybrid map of the human genome.

We have developed a panel of whole-genome radiation hybrids by fusing irradiated diploid human fibroblasts with recipient hamster cells. This panel of 168 cell lines has been typed with microsatellite markers of known genetic location. Of 711 AFM genetic markers 404 were selected to construct a robust framework map that spans all the autosomes and the X chromosome. To demonstrate the utility of the panel, 374 expressed sequence tags (ESTs) previously assigned to chromosomes 1, 2, 14 and 16 were localized on this map. All of these ESTs could be positioned by pairwise linkage to one of the framework markers with a LOD score of greater than 8. The whole genome radiation hybrid panel described here has been used as the starting material for the Genebridge4 panel that is being made widely available for genome mapping projects.

A comprehensive genetic map of the human genome based on 5,264 microsatellites.

The great increase in successful linkage studies in a number of higher eukaryotes during recent years has essentially resulted from major improvements in reference genetic linkage maps, which at present consist of short tandem repeat polymorphisms of simple sequences or microsatellites. We report here the last version of the Généthon human linkage map. This map consists of 5,264 short tandem (AC/TG)n repeat polymorphisms with a mean heterozygosity of 70%. The map spans a sex-averaged genetic distance of 3,699 cM and comprises 2,335 positions, of which 2,032 could be ordered with an odds ratio of at least 1,000:1 against alternative orders. The average interval size is 1.6 cM; 59% of the map is covered by intervals of 2 cM at most and 1% remains in intervals above 10 cM.

An STS-based map of the human genome.

A physical map has been constructed of the human genome containing 15,086 sequence-tagged sites (STSs), with an average spacing of 199 kilobases. The project involved assembly of a radiation hybrid map of the human genome containing 6193 loci and incorporated a genetic linkage map of the human genome containing 5264 loci. This information was combined with the results of STS-content screening of 10,850 loci against a yeast artificial chromosome library to produce an integrated map, anchored by the radiation hybrid and genetic maps. The map provides radiation hybrid coverage of 99 percent and physical coverage of 94 percent of the human genome. The map also represents an early step in an international project to generate a transcript map of the human genome, with more than 3235 expressed sequences localized. The STSs in the map provide a scaffold for initiating large-scale sequencing of the human genome.

Lack of additive effect between mechanisms of resistance to carbapenems and other beta-lactam agents in Pseudomonas aeruginosa.

Eighty-nine clinical isolates resistant (n = 61) or susceptible (n = 28) to imipenem and exhibiting the main patterns of susceptibility to other beta-lactam agents (wild type pattern, penicillinase pattern, constitutive cephalosporinase pattern) were studied in order to investigate (i) the mechanism of resistance involved and (ii) whether resistance to carbapenems affects the level of resistance to other beta-lactam agents and, conversely, if resistance to other beta-lactam agents affects the level of resistance to carbapenems. For this purpose, the presence of OprD protein in the cell wall was detected by Western blot and beta-lactamase activity by spectrophotometric assay and isoelectric focusing. OprD expression was not detectable in the imipenem-resistant (MIC > or = 16 micrograms/ml) strains. It was decreased in half the strains for which MICs of imipenem were 2 to 8 micrograms/ml and was close to a normal level in the most susceptible strains (MIC < or = 1 microgram/ml), thus demonstrating a direct correlation between the level of susceptibility to imipenem and the level of OprD expression. No imipenemase activity was detected in imipenem-resistant strains. Synergy between imipenem or meropenem and BRL 42715 was observed for all of the strains, demonstrating the role of cephalosporinase in carbapenem resistance. Within each pattern of susceptibility, the mean MICs of beta-lactam agents other than carbapenems were similar, whether the strains were susceptible or resistant to imipenem. Conversely, the mean MICs of imipenem or meropenem for either the imipenem-resistant or the imipenem-susceptible strains were similar, regardless of the susceptibility of these strains to the other beta-lactam agents. Thus, when several mechanisms of resistance to beta-lactam agents are present in the same strain of Pseudomonas aeruginosa, there is no additive effect between these mechanisms.

Role of mutations in DNA gyrase genes in ciprofloxacin resistance of Pseudomonas aeruginosa susceptible or resistant to imipenem.

In Pseudomonas aeruginosa, resistance to imipenem is mainly related to a lack of protein OprD and resistance to fluoroquinolones is mainly related to alterations in DNA gyrase. However, strains cross resistant to fluoroquinolones and imipenem have been selected in vitro and in vivo with fluoroquinolones. We investigated the mechanisms of resistance to fluoroquinolones in 30 clinical strains of P. aeruginosa resistant to ciprofloxacin (mean MIC, >8 micrograms/ml), 20 of which were also resistant to imipenem (mean MIC, >16 micrograms/ml). By immunoblotting, OprD levels were markedly decreased in all of the imipenem-resistant strains. Plasmids carrying the wild-type gyrA gene (pPAW207) or gyrB gene (pPBW801) of Escherichia coli were introduced into each strain by transformation. MICs of imipenem did not change after transformation, whereas those of ciprofloxacin and sparfloxacin dramatically decreased (25- to 70-fold) for all of the strains. For 28 of them (8 susceptible and 20 resistant to imipenem), complementation was obtained with pPAW207 but not with pPBW801. After complementation, the geometric mean MICs of ciprofloxacin and sparfloxacin (MICs of 0.3 microgram/ml and 0.5 microgram/ml, respectively) were as low as those for wild-type strains. Complementation was obtained only with pPBW801 for one strain and with pPAW207 and pPBW801 for one strain highly resistant to fluoroquinolones. These results demonstrate that in clinical practice, gyrA mutations are the major mechanism of resistance to fluoroquinolones even in the strains of P. aeruginosa resistant to imipenem and lacking OprD, concomitant resistance to these drugs being the result of the addition of at least two independent mechanisms.