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Object: To determine the frequency, clinical features, and morbidity of Mycoplasma pneumoniae infections., Method: Retrospective study of 76 consecutive children under 16 years of age hospitalized at the Reims University Hospital from 1999 to 2005 with M. pneumoniae pneumonia. The infection was defined by the presence of M antibodies and/or an increase in G antibodies (quantitative Elisa test)., Results: M. pneumoniae was the cause of 16% (76/464) of hospitalized pneumonia cases. A significantly increased frequency was observed in 2004 (34%; 19/56) and 2005 (26%; 22/84) versus 11% from 1999 to 2003, p<5.10(-4). The mean age of the patients was 6 years and 8 months, with a peak at 3 years (14/76; 18% of patients). The most frequent clinical feature was cough (80%; 56/70). The chest X-ray showed typical radiological features such as peribronchial and perivascular interstitial infiltrates in only 23% (16/69). Respiratory and extrarespiratory complications were seen in 17 and 12 children, respectively. Only 1 child suffered from respiratory sequelae., Conclusion: M. pneumoniae pneumonia is frequent in children over 2 years of age. The diagnosis is sometimes difficult to initially assert because there are no specific features. Respiratory and extrarespiratory complications remain possible. Respiratory sequelae can still exist even if most cases evolve favorably under treatment by macrolides.

Background: Human rhinoviruses and enteroviruses (Picornaviridae) are suspected to be major viral etiological causes of bronchiolitis in infants., Objectives: In the present study, we assessed the potential role of the respiratory picornaviruses as causative agents of bronchiolitis in French infants., Study Design: From September 2001 to June 2002, we prospectively selected 192 infants < or =36 months of age and hospitalized for acute bronchiolitis. The detection of common respiratory viruses (respiratory syncytial virus, influenza virus A and B, parainfluenza virus 1, 2, 3 and adenovirus) was performed using classical immunofluorescence antigen and cell-culture detection assays on nasopharyngeal aspirates whereas the detection of human metapneumovirus (HMPV) was performed by a real-time RT-PCR assay. The presence of rhinovirus and/or enterovirus was assessed in respiratory samples by a picornavirus RT-PCR detection assay followed by a differential Southern blotting procedure., Results: A potential causative virus was detected in 72.5% of the 192 study infants. RSV (30%), rhinovirus (21%), enterovirus (9%), influenza virus A (6%) and human metapneumovirus (4%) were the most frequent causative agents detected. Rhinoviruses or enteroviruses were detected as the only evidence of respiratory viral tract infection in 57 (30%) of 192 infants, whereas rhinovirus or enterovirus occurred in mixed viral infection detected in 25 (13%) of 192 study cases (30% versus 13%, p<10(-3))., Conclusions: Our data suggest that respiratory picornaviruses are one of the leading etiological causes of bronchiolitis in French infants. These findings highlight the need to implement a rapid picornavirus RT-PCR detection assay for the clinical diagnosis of respiratory infections in pediatric patients with bronchiolitis.

Influenza pneumonia and influenza-associated severe exacerbation of pre-existing heart and lung disease are responsible for major complications that may require intensive care unit admission. Here, we report the case of a diabetic 70 year-old man hospitalised in the intensive care unit (ICU) of the University Medical Center of Reims (France) for a severe bilateral and alveolar pneumonia requiring mechanical ventilation. This patient had received a classical antibiotic treatment by amoxycillin (3 g/24 hours per os); 48 hours later, he was admitted in ICU for a respiratory failure that evolved rapidly towards an acute respiratory distress syndrome. Because of the context of a winter influenza outbreak, a nasal swabbing sample was tested for the presence of Influenzavirus nucleocapsid-antigens (Immunochromatographic test; BinaxNow Flu A & B, Binax, Portland, USA). This rapid assay revealed the presence of an Influenzavirus A respiratory infection five days after the beginning of the respiratory syndrome. This rapid viral diagnosis will be further confirmed in post mortem by the positive Influenza strain isolation onto lung tissues by classical cell culture techniques (Influenzavirus A strain, H3N2). Influenza pneumonia is a significant cause of morbidity and mortality, especially during influenza epidemics. The use of commercially available rapid diagnostic tests for influenza associated pneumonia, allows the potential use of new specific anti-neuraminidase drugs, which can be efficient during the 30 hours after the beginning of the clinical influenza syndrome.

We described the natural polymorphism of cytomegalovirus DNA polymerase in 42 unrelated isolates susceptible to ganciclovir, foscarnet, and cidofovir. All variations, including an eight-amino-acid deletion, were located between domains delta-C and II and between domains III and I, suggesting that these specific residues are not involved in enzymatic functions.

Introduction: The aim of this study was to assess the prevalence of sexually transmitted infections among patients attending an anonymous HIV Screening Center., Patients and Methods: This prospective study was performed in the HIV Screening Center of University hospital in Reims (France) from May 1997 to December 1997. The inclusion criteria were the asymptomatic clinical presentation and the presence of risk factors for sexually transmitted infections referring to WHO criteria. The methods included clinical examination after application of acetic acid and urethral and endocervical swabs to identify:Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, Gardnerella vaginalis, Trichomonas vaginalis in specific culture. Treponema pallidum and HIV-1 infection were both detected by Enzym Linked Immuno Sorbent Assay (ELISA)., Results: One hundred and one patients (62 men and 39 women) were included in the study. Their mean age was 27 +/- 4 Years. Risk factors for sexually transmitted infections were: multiple sexual partners 81 p. 100; homo or bisexuality 16 p. 100; intravenous drug use 3 p. 100. The sexually transmitted infections were: HIV-1 infection 1 p. 100;Ureaplasma urealyticum 25 p. 100; genital warts 5 p. 100;Chlamydia trachomatis 3 p. 100; Gardnerella vaginalis 3 p. 100; Mycoplasma hominis 2 p. 100; Treponema pallidum 0 p. 100; Neisseria gonorrhoeae 0 p. 100; Trichomonas vaginalis 0 p. 100. The prevalence of sexually transmitted infections was significantly higher among women (p<0.05)., Discussion: Classical sexually transmitted infections and HIV infection were rarely detected in this study; but prevalence of other sexually transmitted infections (genital warts, Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum) was high. Ureaplasma urealyticum is considered as a possible pathogenic agent in pregnant women (preterm delivery, decrease of birth weight, chorioamniotitis). These results suggest that other than sexually transmitted infections in high risk patients attending a HIV Screening Center other sexually transmitted infections should also be systematically screened for.

Enterovirus (EV) detection by a new commercially available reverse transcription (RT)-PCR assay (Penter RT-PCR test) was compared with EV isolation from cell cultures and with EV detection by an in-house RT-PCR assay. Of the 54 cerebrospinal fluid specimens collected during a summer outbreak of aseptic meningitis, 52% were positive by cell culture versus 76% by in-house RT-PCR assay and 80% by the new RT-PCR test (52 versus 76 versus 80%; P = 0.003). This new reliable EV RNA detection test is suitable for clinical diagnosis of EV-related meningitis and may improve the management of EV-related neurological syndromes.

The relationship between a cytomegalovirus (CMV) infection and the acute rejection of a renal transplant is not well established. The aim of the study was to document whether the clinical presentation of a CMV infection as a diffuse inflammatory disease or as a clinically asymptomatic illness is a risk factor of acute renal transplant rejection. One hundred and ninety-two consecutive renal transplant recipients were included in a historical cohort study for exposed-non exposed analyses. CMV infection after transplantation was the exposure factor. Before transplantation, 113 patients had antibodies against CMV and 79 were seronegative. The patients were divided into three groups: Group 1 consisted of 64 patients who had neither clinical signs of CMV disease nor CMV serological changes after transplantation, Group 2 consisted of 77 seropositive patients with asymptomatic viremia, and Group 3 consisted of 51 seropositive patients with clinical signs of diffuse inflammation that included fever, neutropenia, and various visceral involvements (CMV disease). Groups 2 and 3, the seropositive patients, were paired with Group 1 patients. Acute rejection was considered as CMV-induced when it occurred within one month following viremia, during the first year after transplantation. Transplant patients with CMV disease, had a significant likelihood of developing acute rejection after CMV infection or reactivation (P < 0.01). The odds ratio for developing rejection was 5.98, 95% confidence interval: 1.21-29.40. Such a link was not documented for recipients with asymptomatic CMV infection. In conclusion, CMV disease, but not asymptomatic viremia, is a risk factor of acute renal transplant rejection. On epidemiological grounds, these results support the hypothesis that factors controlling both the viral replication and the diffuse inflammatory process are implicated in acute graft rejection.

Objectives: We studied the relationship between cytomegalovirus infection and episodes of acute rejection after infection in renal graft recipients at the Reims University Hospital from 1989 to 1995., Patients and Methods: Two exposed versus nonexposed analyses were conducted, one (series 1) for CMV infection and the other (series 2) for CMV disease. For each analysis, exposed recipients were matched with nonexposed recipients for date of graft (+/- 6 months). Risk of acute rejection was assessed with univariate analysis then with multivariate analysis using logistic regression., Results: Among the 192 graft recipients included, 64 developed CMV infection, 77 had an infection (series 1) and 51 had CMV disease (series 2). In series 1, only failure of renal graft was a significant risk factor of acute rejection (OR = 10.4; 95% Cl 1.9-56.3). CMV infection was not a significant risk factor (OR = 1.06; 95% Cl 0.2-5.6). Conversely, in series 2, there was a 6-fold increase in the risk of acute rejection in recipients who developed CMV disease (OR = 5.98; 95% Cl 1.21-29.4)., Conclusion: The fact that CMV disease and not CMV infection is a risk factor of acute rejection in renal transplant recipients is an argument for implicating a general inflammatory reaction characteristic of CMV disease in the pathogenesis of acute rejection. This finding favors preventive treatment of CMV infection.

Cidofovir (CDF) or Vistid is a monophosphate nucleoside analogue that inhibits the DNA polymerase of herpes viruses including the cytomegalovirus (CMV). CDF is active on GCV-resistant strains with a mutation on the phosphotransferase gene (UL97). However, DNA polymerase gene mutations that induce resistance to GCV are responsible for cross-resistance to CDF. Resistance phenotypes to GCV and CDF were determined for 57 CMV strains isolated from blood and urine samples. Sixteen strains were recovered after CDF therapy. Of the remaining 41 CDF-naive strains, 34 were susceptible and seven resistant to GCV. Fifty percent inhibitory concentrations (IC50) for CDF were in the 0.2-2.6 microM range for CDF-naive strains susceptible to GCV. For GCV-resistant strains, IC50 values for CDF were < or = 3 microM for strains with a low level of resistance to GCV (GCV IC50 < 30 microM) and > or = 6 microM for three of the five strains with a high level of resistance to GCV (GCV IC50 > or = 30 microM).

The substitution of methionine by either isoleucine or valine at residue 460 in the UL97 phosphotransferase has been shown to be responsible for resistance to ganciclovir (GCV) in 30% of resistant cytomegalovirus (CMV) isolates [4]. These substitutions require one nucleotide change in the gene (G- > T 1 380 and A- > G 1378 respectively). The aim of this study was to develop a discriminative PCR assay for rapid detection of these DNA changes. A PCR assay was duplicated in parallel for each mutation; to detect G- > T 1380 each reaction mixture contained primer VSUL14 and either primer LNW to distinguish wild type residues or LNM to distinguish mutant residues, and for A- > G 1378 primers were VSUL8 and either MCMW to detect wild type sequences or MCMM to detect mutated residues. For optimal discrimination, primers MCMW and MCMM were designed with a mismatch at position 3'-1. The reference strains AD169, Davis and Towne, a laboratory GCV-resistant mutant RCL1.7, and 33 CMV isolates (10 resistant, 2 indetermined and 21 sensitive) were tested by PCR. AD169, Davis and Towne, and 30 isolates were amplified only with non modified primers, and the absence of 460 mutations was confirmed by sequencing. Two isolates P1 and P2, from a transplanted patient were amplified with both MCMM and MCMW: sequencing analysis shown the presence of a mixture of strain, one of them harbouring A- > G 1378 mutation. One resistant strain was amplified neither with MCMM nor with MCMW: a C- > T silent mutation at nt 1368 was present. As sequencing analysis confirmed PCR results, discriminative PCR enables isolates to be rapidly assessed for the presence or absence of 460 mutations. Moreover, it can distinguish Met to Val from Met to Ile mutations, and allows the analysis of mixtures of sensitive ad resistant strains.

Bacterial adhesion to biomaterials is a complex phenomenon involving numerous factors. The ability to reduce urinary catheters infections simply by general hygiene and asepsis is low: an ascending colonization cannot be avoided. This will lead to a clinical infection only if several factors favour the bacterial adhesion or the bacterial coaggregation and the feeding of the bacterial biofilm. Among the many factors involved in bacterial adhesion, we focused in this paper on the physical parameters of surface hydrophobicity of the urinary catheters (Van der Waals and acido-basic forces) and the surface hydrophobicity of the bacteria (BATH and zeta potential). We also compared scanning electron microscopy (SEM) of in vivo and in vitro infected urinary catheters. We provided evidence that the more hydrophobic the bacteria, the more they are able to colonize hydrophobic materials, whereas hydrophilic cells are able to colonize hydrophilic materials more easily. Some biomaterials were found to display an irregular texture of hydrophobic and hydrophilic areas: they favour both types of adhesion. Moreover the divalent cations (MgII) drastically increased the bacterial coaggregation and favour bacterial growth within the biofilm. Finally, an increase in urinary pH and ionic strength increases the colonization risk. Consequently, choice of urinary catheter biomaterials is essential as patient hygiene and diet in order to avoid clinical infections.

Cariogenic dental plaque may be assimilated to a biofilm resulting from the adhesion of S. mutans, then from the coaggregation of other streptococci, or other genus. We used a static monospecific biofilm model. Supports or bacteria were treated with inhibitors before adhesion in order to clarify the nature of adhesins responsible for the primary adhesion of S. mutans and S. sobrinus on Tygon. To determine the bindings of coaggregation, inhibitors were applied on one-day-old biofilms. Analysis of effects were performed by automatic inoculator Spiral (Interscience) for microbiological methods, and by SEM JEOL 5400 LV for microscopic methods. In the aim of preventing adhesion and coaggregation, different traps were assayed:sugars, chemical inhibitors such as F- and EDTA salts. Of these, only the latter showed efficiency. This confirmed the role of bivalent mineral ions and electrostatic attraction forces in the adhesion and coaggregation of streptococci.

Disrupting bacterial biofilms is necessary for a wide application domains such as reusable medical devices, or systems of pipes for water or fluids in cosmetics, food and chemicals industry. Bacterial cells embedded in a biofilm are less susceptible to disinfectants than suspended cells. This property is referable to the structure of the biofilm itself. The gangue of exopolymers and the thickness of a 5-day-old biofilm of Escherichia coli (more than 200 layers of bacteria), contribute to this decrease of susceptibility. The present work deals with the release of an Escherichia coli biofilm by the sequential action of enzymes and a phenolic disinfectant on the one hand, and by the sequential or simultaneous action of surfactants and the previous disinfectant on the other hand. The decrease of bacteria count per mm2 and the Scanning Electron Microscope observations exhibited a synergic action in every case. Nevertheless, Escherichia coli biofilms quickly reconstructed even after exposition to the previous treatment.

S. mutans is one of the bacterial species involved in the initiation of cariogenic dental plaque. In situ, antiseptic activity evaluation of dental plaque chemicals is fairly difficult. A dynamic biofilm model of S. mutans on Tygon support is suitable for the quantification of antiseptic activity and the comparison of commercial chemical formulations, submitted or not to the french standards and Pharmacopea. Some of these chemicals, especially quaternary ammonia solutions, do not reduce living bacteria counts by 10E5 as AFNOR specifies. In the case of dental plaque, antiseptic activity is probably not the only relevant parameter: biofilm dissociation is rather more relevant because the dead or living bacteria adhere to the teeth, favouring the establishment of a new active biofilm. Two proceedings can be followed: support surface can be modified, or bacteria exopolymers synthesis can be inhibited to prevent cell adherence; the disruption of the biofilm glycocalyx can be achieved with enzymes or surfactants.

Cytomegalovirus (CMV) is the most common opportunistic pathogen following renal transplantation and remains a major concern in transplantation centers owing to its high morbidity and impact on renal allografts. Pending more effective antiviral drugs, efforts have been directed toward prevention strategies. We conducted a retrospective analysis to evaluate the efficacy of various prophylactic options used at our institution during the period April 1986 to August 1990. All CMV-negative patients with CMV-negative kidneys (D-R-) received screened, CMV-negative blood products (n = 19). CMV-specific immunoglobulins (CMV Ig) were used in 6 patients at increased risk for primary CMV infection and acyclovir was administered to 21 patients at an initial intravenous dose of 5 mg/kg body weight; then oral doses of 800-3200 mg per day were given according to the patients' estimated creatinine clearance. Thirty-two patients did not receive any CMV prophylactic treatment and served as controls. CMV monitoring of the patients during the first 6 months after transplantation showed an overall infection and disease rate of 81% and 38.1%, respectively, in the acyclovir-treated group. Compared with controls, the incidences of infection and disease were higher in the acyclovir-treated patients, with a significant difference for CMV infection (P = 0.002, generalized Wilcoxon test). Only 1 of the 19 D-R- patients presented with CMV infection. CMV Ig-treated patients tended to have less severe disease without any apparent reduction in infection incidence. Given the high rate of infection in patients at risk, we infer that high-dose acyclovir does not prevent CMV infection in our setting of renal transplantation.(ABSTRACT TRUNCATED AT 250 WORDS)

When bacteria colonize a surface they form a biofilm whose susceptibility to anti-microbials is different from that of the same bacterial species forming a homogeneous suspension in a liquid. This study investigated colonization of an inert solid phase (Tygon) with Streptococcus mutans ATCC 25,175--one of the strains involved in the initiation of cariogenic dental plaque--in a continuous flow of fresh medium sufficiently diluted so as to preclude growth of suspended bacteria. Only those bacteria which adhered to the solid phase grew, forming a biofilm. The antiseptic activity of Eludril (0.1% chlorhexidine) on this biofilm under dynamic (flowing medium) and static (stagnant medium) conditions was studied by comparison with the same strain in a suspension (in compliance with the AFNOR NF T 72-150 norm) and in a confluent culture on a filtering membrane. The biofilm was less susceptible under dynamic than under static conditions; under both conditions, the biofilm was less susceptible than the suspension. According to this model, the concentration of antiseptic recommended by the manufacturer according to studies using AFNOR norm NF T 72-150 (bacteria in a suspension) may be inadequate for bacteria adhering to tooth surfaces or gingival mucosa.

A case of fatal cytomegalovirus pneumonia in a non-immunocompromised 65-year old man is reported. The patient presented with symptoms of a lower respiratory tract infection. The diagnosis of cytomegalovirus pneumonia was made after histopathological examination of the open lung biopsy. Confirmation of cytomegalovirus infection was made with in situ DNA Hybridization. The outcome was rapidly fatal. This case demonstrates the value of DNA probe analysis for diagnosis. This case is also unusual in that a non-immunocompromised adult had severe evolution of cytomegalovirus infection.

Antibodies against Epstein-Barr virus (EBV) antigens, i.e. the viral capsid antigen (VCA) and the Epstein-Barr nuclear antigen (EBNA), were determined in two independent populations with known HLA-DR phenotypes. The first population consisted of 151 patients with rheumatoid arthritis; the second one of 88 healthy parents of leukemic children. Although the group of patients with rheumatoid arthritis differed significantly in the frequency of 4 DR antigens from the second group, both groups had the same correlation between HLA-DR antigens and the antibody response to EBV antigens. There was a significant correlation between HLA-DR1 and reduced titers of antibodies to VCA, whereas the persons with only one identifiable DR antigen had higher anti-VCA titers. The persons with HLA-DR5 had significantly higher anti-EBNA titers than those without DR5.