Unexpected mould diversity in clinical isolates from French Guiana and associated identification difficulties.

New mold species are increasingly reported in invasive fungal infections. However, these fungi are often misdiagnosed or undiagnosed due to the use of inappropriate laboratory diagnostic tools. Tropical countries, such as French Guiana, harbor a vast diversity of environmental fungi representing a potential source of emerging pathogens. To assess the impact of this diversity on the accuracy of mold-infection diagnoses, we identified mold clinical isolates in French Guiana during a five-month follow-up using both microscopy and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. In total, 38.8% of the 98 obtained molds isolates could not be identified and required a DNA-based identification. Fungal diversity was high, including 46 species, 26 genera, and 13 orders. Fungal ecology was unusual, as Aspergillus species accounted for only 27% of all isolates, and the Nigri section was the most abundant out of the six detected Aspergillus sections. Macromycetes (orders Agaricales, Polyporales, and Russulales) and endophytic fungi accounted for respectively 11% and 14% of all isolates. Thus, in tropical areas with high fungal diversity, such as French Guiana, routine mold identification tools are inadequate. Molecular identifications, as well as morphological descriptions, are necessary for the construction of region-specific mass spectrum databases. These advances will improve the diagnosis and clinical management of new fungal infections. LAY SUMMARY: In French Guiana, environmental fungal diversity may be a source of emerging pathogens. We evaluated microscopy and mass spectrometry to identify mold clinical isolates. With 39% of unidentified isolates, a region-specific mass spectrum database would improve the diagnosis of new fungal infections.

Prosthetic treatment in the adult French population: Prevalence and relation with demographic, socioeconomic and medical characteristics.

BACKGROUND: The aim of this study was to estimate the prevalence of dental prosthetic treatment and to investigate the demographic, social, economic and medical factors associated with the use of fixed and removable dentures in a representative sample of adults living in France. METHODS: The data were obtained from the 2002-2003 Decennial Health Survey, a cross-sectional study of a representative sample of the population living in France, which included 29,679 adults. Information was collected by interview. The variables collected were fixed denture, removable denture, age, gender, number of children, area of residence, nationality, educational attainment, family social status, employment status, annual household income per capita, supplementary insurance, chronic disease, eyesight problems/glasses, hearing problems/hearing aids. Multinomial logistic regression models were used to study the relationship between prosthetic treatment and demographic, socioeconomic and medical characteristics unadjusted, adjusted for age and adjusted for all the characteristics. RESULTS: The prevalence of prosthetic treatment was 34.6% (95% confidence interval (CI): [34.1; 35.2]) for fixed prosthetic dentures and 13.8% (95% CI: [13.4; 14.2]) for removable prosthetic dentures. We showed a gradient between educational attainment and removable dentures; the odds ratio adjusted for all the variables (aOR) associated with no or primary education compared to post-secondary education was 2.56; 95% CI: [2.09; 3.13]. When annual household income per capita was low, subjects were less likely to report fixed dentures (aOR=0.68; 95% CI: [0.62; 0.75]) than those with high annual household income per capita. Individuals without insurance less often reported fixed dentures than those with private insurance. Those reporting chronic disease were less likely to report fixed dentures (aOR=0.87; 95% CI: [0.79; 0.95]) but more likely to report removable dentures (aOR=1.29; 95% CI: [1.17; 1.43]) than those without chronic disease. CONCLUSION: This study reveals social, economic and medical inequalities in fixed and removable prosthetic treatment among adults in France.

Histoplasma capsulatum causing sinusitis: a case report in French Guiana and review of the literature.

BACKGROUND: American histoplasmosis is a mycosis caused by Histoplasma capsulatum. A variety of clinical features of histoplasmosis have been commonly described ranging from asymptomatic infections to severe pulmonary infections. In immunocompromised individuals, progressive disseminated forms are frequent, leading to fatal outcome if untreated. However, H. capsulatum sinusitis is unusual with a few cases documented over the last three decades and may be underdiagnosed or confused with other fungal aetiologies, especially outside endemic regions. CASE PRESENTATION: In this study, we report an atypical case of Histoplasma capsulatum sinus fungus ball-like in a patient with Acquired Immunodeficiency Syndrome due to Human Immunodeficiency Virus complicated by a disseminated histoplasmosis with a death ending. Diagnosis relied on CT-Scan imaging and on both direct examination of H. capsulatum yeast forms (Gomory methenamine Grocott) in the sinus specimen (aspirate, biopsy) and on positivity of the culture further confirmed by qPCR. CONCLUSIONS: Since last few decades, among the eight reviewed patients, H. capsulatum sinusitis occurred mostly in HIV-immunocompromised patients and for three cases as a sinusitis with disseminated histoplasmosis. Even if this is a rare clinical presentation, its diagnosis is crucial as it could represent an early expression of an Histoplasma capsulatum exposure that can evolve into a disseminated fatal infection when immunity decreases.

Fusarium volatile, a new potential pathogen from a human respiratory sample.

We describe the isolation and characterization of Fusarium volatile from a bronchoalveolar lavage (BAL) sample of a female patient living in French Guiana with underlying pulmonary infections. Phylogenetic analysis of fragments of the calmodulin (cmdA), translation elongation factor (tef1), RNA polymerase second largest subunit (rpb2), and ß-tubulin (tub) loci revealed that strain CBS 143874 was closely related to isolate NRRL 25615, a known but undescribed phylogenetic species belonging to the African clade of the Fusarium fujikuroi species complex. The fungus differed phylogenetically and morphologically from related known species, and is therefore described as the new taxon Fusarium volatile. Antifungal susceptibility testing suggested that the new species is resistant to echinocandins, fluconazole, itraconazole with lower MICs against amphotericin B, voriconazole and posaconazole.

Selective CD4+ T-cell depletion does not prevent graft-versus-host disease.

Donor-derived CD4+ T cells may play a role in the development of graft-versus-host disease (GVHD) and graft-versus-leukemia reaction after allogeneic bone marrow transplantation (BMT). Therefore, we evaluated the effect of CD4+ T-cell depletion on GVHD and graft-versus-leukemia reaction after HLA-matched BMT. CD4 depletion was performed using anti-CD4 monoclonal antibodies and immunomagnetic beads, initially in small-scale experiments on bone marrow and granulocyte colony-stimulating factor-mobilized peripheral blood apheresis products. The result was elimination of the CD4+ T cells from both sources (0% and 2+/-1.4% CD4+ cells, respectively). Subsequently, we used this technique for large-scale negative selection of CD4+ T cells from bone marrow grafts of four consenting leukemic patients in relapse (ALL-3, ANLL-1) (M-3, F-1). The large-scale CD4+ T-cell depletion resulted in >98% (n=4) elimination of CD4+ cells. The resulting population included 17.7+/-4.6% CD3+ T cells, 8.9+/-2.5% CD8+ T cells, 0.1+/-0.1% CD16+ natural killer cells, and 2.3+/-3.2% CD34+ hematopoietic progenitor cells. Patients were transplanted with 2.84+/-1.31 x 10(8) viable cells/kg. They received cyclosporine starting on day -1 as GVHD prophylaxis. Engraftment was fast with a white blood cell count of >1 x 10(9)/L on day 13.2+/-0.5, an absolute neutrophil count of >0.5 x 10(9)/L on day 13.8+/-0.5, and a platelet count of >25 x 10(9)/L on day 26.5+/-6.8. Immunological reconstitution was normal, and peripheral blood phenotyping 3 weeks after BMT disclosed 49.0+/-5.0% CD3, 14.3+/-12.4% CD4, and 59.5+/-7.8% CD8 T cells in addition to 17.0+/-3.0% CD16+ and 9.0+/-3.0% CD56 natural killer cells. Three out of four patients developed very early grade IV GVHD beginning on day 12 (10-13) and died 2-4 months after BMT. One patient is alive and well with a follow-up of 36 months. We conclude that selective CD4 T-cell depletion does not prevent GVHD.

The effect of linomide, an immunoregulator in experimental autoimmune diseases, on humoral antibody responses in mice.

Linomide (quinoline-3-carboxamide), a well tolerated, orally administered compound was recently shown to be effective in the prevention and treatment of several autoimmune diseases in experimental animal models. We have investigated its effect on specific humoral immune responses directed to T-cell-dependent soluble or particulate antigens and to a T cell-independent antigen in several mouse strains. Linomide administered after antigen priming did not affect primary and secondary antibody responses directed to T-cell particulate antigens (SRBC) or soluble antigens given with or without complete Freund's Adjuvant (CFA). Linomide treatment given prior to antigen priming did not affect the antibody response to a soluble antigen (TNP-KLH) given with an adjuvant. In contrast, dose-dependent down regulation of primary antibody responses was observed when T cell-dependent (BSA-dextran) or T-cell-independent (TNP-Ficoll) antigens were administered in an immunogenic form without adjuvant after starting Linomide treatment. The primary anti-SRBC antibody response was also suppressed by high dose Linomide given prior to immunization although normal secondary responses were retained. It is worth noting that no immunosuppressive effects on antibody responses were found at low dose ranges which effectively reversed T cell dependent autoimmune manifestation.

The effect of in vitro T lymphocyte depletion on generation of IL2-activated cytotoxic cells.

The effect of immunocompetent lymphocyte depletion on precursors and effector cells of IL2 activated non-MHC restricted cytotoxic cells (LAK) generated from bone marrow (BM) or peripheral blood was investigated. Lymphocyte depletion was carried out by using Campath-1, a monoclonal rat anti-human lymphocyte antibody recognizing CDW52, that binds human complement and is used routinely in clinical bone marrow transplantation (BMT). The results indicate that LAK precursors derived from BM cells are sensitive to Campath-1 treatment, while a variable degree of sensitivity was demonstrated in LAK precursor cells derived from peripheral blood. In contrast, effector LAK cells generated in vitro were shown to be resistant to treatment with Campath-1 and complement. We hypothesize that if indeed IL2-dependent non-MHC restricted cytotoxic cells play a role in vivo in the immediate post-BMT period, a T lymphocyte depletion procedure such as the Campath-1 may have the capacity to reduce, at least temporarily, the graft-versus leukemia effects mediated by such anti-tumor effector mechanisms.

Activated allogeneic cell therapy (allo-ACT) for relapsed chronic myelogenous leukemia (CML) refractory to buffy coat transfusions post-allogeneic bone marrow transplantation.

We describe a 17-year-old male patient with chronic myelogenous leukemia (CML) in hematologic and cytogenetic relapse 4 months post-non-T cell-depleted allogeneic bone marrow transplantation for accelerated CML. Two sequential buffy coat transfusion with donor peripheral blood cells (8.9 and 4.8 x 10(7) cells/kg), the second transfusion in combination with in vivo activation of donor cells by human recombinant interleukin-2 (rIL-2) 6 x 10(6) IU/m2 subcutaneously for 3 days, failed to induce remission . The patient responded to an infusion of donor peripheral blood lymphocytes (3.4 x 10(7) cells/kg) pre-activated in vivo with rIL-2 and additionally activated in vivo with rIL-2, 6 x 10(6) IU/m2/day subcutaneously for 3 days. Elimination of the Philadelphia (Ph) clone was confirmed by cytogenetic analysis showing a normal male karyotype and by disappearance of the bcr/abl transcript, using the polymerase chain reaction (PCR). At present, the patient is 26 months post-treatment with no evidence of disease, but with chronic graft-versus-host disease. Our data indicate that allogeneic activated cell therapy (allo-ACT) may provide antitumor effector cells that successfully induce graft-versus-leukemia (GVL) effects even when cell therapy with donor buffy coats was insufficient.

Active and inactive caries lesions in a selected elderly institutionalised French population.

Various epidemiological investigations have shown the high prevalence and incidence of caries in geriatric populations. The aim of the present study was to evaluate the dental status and some salivary parameters of elderly French subjects institutionalised in a geriatric hospital. The study population included 117 subjects (31 males and 86 females) who were neither demented nor edentulous, with a mean age of 83.0 years (SD = 7.8). Crown and root caries were recorded according to a modified caries activity index on 17,442 surfaces (9 surfaces per tooth: 5 crown surfaces and 4 root surfaces). The 17,442 surfaces examined corresponded to 1,938 teeth. The mean number of teeth per subject was 16.6 (SD = 7.6), more teeth remaining in the mandible than in the maxilla. The 2,985 unsound root surfaces showed a high percentage of active lesions (31.2 per cent) and a low percentage of filled root surfaces (4.5 per cent). Crowns and roots also presented a high percentage of destroyed surfaces: 1,446 destroyed surfaces, 8.3 per cent of the examined surfaces. Salivary parameters (flow rate and buffer capacity) were also recorded. Stimulated salivary flow rate had a relationship with crown caries (linear regression and analysis of covariance), and buffer capacity with root caries (analysis of variance and covariance). The data illustrate critical treatment needs in French geriatric institutions. This situation, which appears to differ from previous reports in European and US elderly people, may be related to French specificities concerning oral health status and/or care policy, but also to the usually very old population.

Impact of social and economic characteristics on orthodontic treatment among children and teenagers in France.

OBJECTIVES: This study aimed to estimate the prevalence of orthodontic treatment in France among children and teenagers aged 8-18 years, by sex and by age, and to investigate the specific role of social and economic characteristics on use of orthodontic treatment. METHODS: We analyzed data from the cross-sectional national health survey conducted in France in 2002-2003, which included a sample of 5988 children aged 8-18 years. All data were collected by interview including the question on orthodontic treatment. Other data used in our study were family social status and income, maternal educational attainment and place of birth, whether the child was covered by a supplementary health insurance and whether the residence was urban or rural. We also calculated the density of orthodontists in the district. Multivariate logistic regression analyses were used to study the relationships between these social and economic factors and orthodontic treatment. RESULTS: The prevalence of orthodontic treatment was 14% of all children aged 8-18, 15% for girls, and 13% for boys, and 23% in the 12 to 15-year age group. Children were less likely to have orthodontic treatment when parents were service or sales workers compared with children whose parents were managers or professionals (aOR = 0.50; 95%CI: [0.34;0.76]), when family income was in the lowest, compared with highest quartile (aOR = 0,62; 95% CI: [0.45;0.85]), when children had no supplementary insurance compared with children covered by private insurance (aOR = 0.53; 95% CI: [0.34; 0.81]), or when they lived in rural compared with urban areas (aOR = 0.70; 95% CI: [0.54; 0.91]). CONCLUSION: There are social inequalities in orthodontic treatment in France, associated mainly with social status, annual income, supplementary insurance, and the residence area.

Caries activity and associated risk factors in elderly hospitalised population--15-months follow-up in French institutions.

UNLABELLED: Only a few studies have been published concerning hospitalised elderly disabled people. OBJECTIVES: 1) to investigate the oral health status of elderly French patients hospitalised in the two main geriatric hospitals of Paris. 2) to describe the respective influences of general parameters (type of hospitalisation, pathologies and medication) on oral environment parameters. 3) to analyse the influences of these oral parameters on caries activity in Long-Term Care (LTCF) and in rehabilitation facilities (RF) patients and to study the incidence and the time-course of caries in these specific population. SUBJECTS: 117 subjects (mean age = 83.0 years, SD = 7.8, range = 64 to 102 years) were examined at baseline and 32 of the 50 LTCF subjects were reexamined 15-months later. METHODS: The general parameters recorded were age, gender, type of hospitalisation, period of stay, removable prosthesis, general diseases, number of diagnoses, medications with hyposalivary side-effects. The oral environment parameters recorded were flow rate, buffer capacity, mutans streptococci and lactobacilli counts, measured at baseline by tests on stimulated saliva, and plaque index. Crown and root surfaces were recorded according to a modified caries activity index. RESULTS: Among the polypathological subjects (85.5% of the population), the number of diseases ranged from 2 to 8. The LTCF patients had a significantly higher mean number of diagnoses (3.5; SD = 1.5) than the RF patients (2.8; SD = 1.4). 76.9% of patients were taking medications with hyposalivary side-effects. The stimulated flow rate ranged from 0.02 ml/min to 5 ml/min. Its mean was significantly lower for LTCF patients (0.67 ml/min; SD = 0.51) than for RF patients (1.12 ml/min; SD = 0.89). The plaque index was significantly higher in LTCF subjects and in patients with mental diseases. At baseline, 17,442 crown and root surfaces were examined. Flow rate was related to crown caries and buffer capacity to root caries. During the 15-months follow-up, the mean number of active root surfaces was significantly increased: from 0.148 (SD = 0.116) at baseline vs. 0.250 (SD = 0.174) at the second examination. CONCLUSIONS: The strongest relationship in the present study between oral parameters and caries activity was the negative relationship between buffer capacity and active root caries. This study confirms an association between the type of hospitalisation and both salivary parameters flow rate and plaque index. This investigation illustrates the critical need for hygiene and oral care, in this elderly disabled population.

Effect of various cytokine combinations on induction of non-MHC-restricted cytotoxicity.

Efforts were directed to achieve an increased lymphokine-activated non-MHC-restricted killing (LAK) activity greater than that induced by rIL-2 alone. Human peripheral blood (PB) and bone marrow (BM)-derived mononuclear cells (MC) were exposed in vitro to multiple cytokine combinations, including rIL-6, rIL-7, rIFN-alpha and rIFN-gamma in the presence of either suboptimal or optimal doses of rIL-2. Our results have shown that BMMC are a potential source for induction of increased LAK activity upon exposure to multiple cytokine combinations, whereas PBMC could not be successfully stimulated under the same conditions. Fifty-five to 62% of BM-derived samples stimulated with high dose rIL-2 + rIL-7 or rIL-2 + rIL-7 + rIL-6 + rIFN-gamma exhibited a higher degree of cytotoxicity than BM samples stimulated with rIL-2 alone. Exposure of PB-derived large granular lymphocytes (LGL) to various cytokine combinations led to increased proliferation after stimulation with suboptimal dose of rIL-2 in the presence of rIL-6 and rIL-7. This increase was not observed in induction of cytotoxicity. We suggest that BMMC activated by multiple cytokine combinations could play an active role in improving antitumor response in vivo by contributing to the control of minimal residual tumor cell growth, particularly post-BM transplantation.

Activated long-term peripheral blood cultures as preparation for adoptive alloreactive cell therapy in cancer patients.

Different modes of in vitro activation of peripheral blood mononuclear cells (PBMC) were compared for their effect on long-term propagation. PBMC cultures were activated by short exposures to the mitogen phytohemagglutinin (PHA) and the CD3 complex, with or without secondary signals provided by ligands of CD28 costimulatory molecules. Activation and long-term cultures were carried out in the presence of recombinant interleukin-2 (rIL-2). Addition of supernatant derived from IL-2-activated PBMC improved culture cell yield. Cumulative fold expansions ranged between 10(3) and 10(5) within 21 days. The highest cell yield was found after PHA activation. Fewer cells were obtained after activation with a combination of CD3 and CD28, and even fewer were obtained after CD3 activation alone. An increase in CD8+ and CD56+ cells, without change in CD4+ cells, was found in activated cultures when compared with fresh PBMC. Non-MHC-restricted cytotoxic activity was documented in all activated cultures. Cytotoxic activity per culture was highest in PHA-activated PBMC because of the high cell yield on the day of harvest. Successful in vitro expansion of PBMC might be helpful for gene transfer into T lymphocytes, as well as for the induction of an antitumor response, particularly for prevention and treatment of relapse of hematologic malignancies following allogeneic or autologous bone marrow transplantation.

Immunological evaluation of patients with hematological malignancies receiving ambulatory cytokine-mediated immunotherapy with recombinant human interferon-alpha 2a and interleukin-2.

Immunological parameters were evaluated in patients treated with cytokine-mediated immunotherapy (CMI) consisting of low doses of recombinant human interferon alpha 2a (rIFN alpha) and recombinant human interleukin-2 (rIL-2) administered either concomitantly or sequentially by subcutaneous self-injections in an outpatient setting. Twenty-six patients with hematological malignancies and 2 metastatic melanoma patients in a progressive stage were enrolled in this clinical trial. Of the 26 patients, 24 were at a stage of minimal residual disease, including 14 patients who had received autologous bone marrow transplantation (ABMT) 2-5 months previously, 7 chronic myelogenous leukemia (CML) and 3 acute myeloid leukemia (AML) patients. Two patients (1 CML and 1 mult. myeloma) were treated at a stage of progressive disease. Non-MHC-restricted cytotoxicity directed against natural-killer(NK)-resistant (Daudi) and NK-sensitive (K562) target cells was assessed before, during and after CMI, either in fresh peripheral blood samples (spontaneous activity) or after in vitro rIL-2 activation (induced activity). Spontaneous killing activity was low prior to treatment, but increased upon termination of treatment in 10/15 evaluated cycels. rIL-2-activated cytotoxicity in vitro was markedly elevated in 8/12 and 6/8 patients after one and two cycles, respectively, of sequential treatment, as well as in 3/8 CML and 5/6 patients after one and two cycles, respectively, of concomitant treatment. Activation of the T cell mitogenic response was demonstrated in 6/9 patients after concomitant CMI, while no such effect was observed throughout a sequential treatment in lymphoma and leukemia patients after ABMT. Although a direct correlation between immune stimulation and the in vivo antitumor response cannot yet be determined, our clinical observations support a beneficial therapeutic effect in a substantial number of patients. These results indicated that the ambulatory CMI protocol of rIL-2 and rIFN alpha could stimulate the host defense immune system and may be helpful in mediating the in vivo antitumor response in patients with minimal residual disease.