Role of a NSAID in the apparent cure of a fungal mycetoma.

UNLABELLED: We report the case of a fungal mycetoma due to Madurella mycetomatis that failed to respond to surgery and antifungal treatment but responded strongly to the addition of a non-steroidal anti-inflammatory drug (NSAID). This African patient was born in Mauritania in 1972. He was a herdsman, living close to the Senegal River. The first nodules appeared on the left foot at the age of 13years (1985). The patient suffered frequent flare-ups with the appearance of black grains and underwent surgery in 1988 and 1992 in Senegal. After remission for several months after surgery, new fistulae occurred. The patient emigrated to France in 1995 and underwent a third surgical intervention in 1996. M. mycetomatis was cultured from the black grains. The patient was otherwise in good health, with no diabetes, and HIV tests were negative. We saw the patient for the first time in 2005, at which time he had flare-ups every two to three months. Imaging disclosed an absence of bone involvement. The patient underwent a fourth operation in October, 2005, and voriconazole treatment was initiated. A new flare-up occurred in February, 2006. CT, MRI, and PET scans revealed calcaneus and tarsal involvement, and posaconazole then replaced voriconazole. Flucytosine was added four months later, due to an absence of improvement. New flares-ups occurred and a fifth surgical intervention was performed in September, 2006. The pain, which had been present for three years, worsened; the patient had to stop working and was no longer able to walk without crutches. Amputation of the foot was considered. Empiric treatment with a NSAID, diclofenac (Voltaren(®); 100mg/day), was added to the antifungal treatment in November 2006, to treat the patient's pain and inflammation. A major improvement was observed within one week. The patient was able to walk without crutches one month later. After two months, clinical examination was normal: no pain, inflammation, nodules or fistulae. Flucytosine was stopped after six months of treatment, in January 2007, diclofenac after 10months, in October 2007, and posaconazole after 18.5months, also in October 2007. No relapse has occurred during the eight years of follow-up since treatment ended. The patient seems to have been cured and has normal CT, MRI, and PET scans. IN SUMMARY: This eumycetoma, which had progressed over 20years despite surgery and antifungal treatments, seems to have been cured by the addition of a NSAID. This observation suggests that inflammation plays a major role in the pathogenesis of fungal mycetoma. Clinical studies of treatments including an NSAID should be conducted to confirm this finding.

Aspergillus PCR in serum for the diagnosis, follow-up and prognosis of invasive aspergillosis in neutropenic and nonneutropenic patients.

We evaluated the usefulness of a serum Aspergillus PCR assay for the diagnosis and prognosis of invasive aspergillosis in a study involving 941 patients for a total of 5146 serum samples. Fifty-one patients had proven/probable aspergillosis. We compared galactomannan (GM), PCR and mycologic analysis of pulmonary samples in both neutropenic and nonneutropenic patients. PCR performed in serum yielded 66.7% sensitivity, 98.7% specificity, 75.6% positive predictive value and 98.0% negative predictive value, while the GM index yielded 78.4% sensitivity, 87.5% specificity, 27% positive predictive value and 98.6% negative predictive value. The inclusion of PCR in the European Organization for Research and Treatment of Cancer (EORTC) and the Mycosis Study Group (MSG) mycologic criteria permitted the reclassification of nine other cases from possible to probable aspergillosis and increased the sensitivity to 71.7%. Combining the GM index with serum PCR increased the detection rate of invasive aspergillosis with 88.2% sensitivity. PCR was systematically negative in 16 patients with noninvasive forms of aspergillosis (namely aspergilloma and chronic aspergillosis). Remaining PCR positive after a period of 14 to 20 days of treatment was related to poor outcome at 30 and 90 days. Our results also indicate that, unlike the determination of the GM index, the initial fungus load as determined by PCR was highly predictive of 90-day mortality, with the rate of the latter being 15.8% for patients with <150 copies/mL vs. 73.2% for patients at or above that cutoff (p <0.0001). Therefore, PCR appears to be a powerful and interesting tool for the identification of patients with invasive aspergillosis who might benefit from more intense care.

Use of mass spectrometry to identify clinical Fusarium isolates.

Fusarium spp. have recently emerged as significant human pathogens. Identification of these species is important, both for epidemiological purposes and for patient management, but conventional identification based on morphological traits is hindered by major phenotypic polymorphism. In this study, 62 strains, or isolates, belonging to nine Fusarium species were subjected to both molecular identification TEF1 gene sequencing and matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) analysis. Following stringent standardization, the proteomic-based method appeared to be both reproducible and robust. Mass spectral analysis by comparison with a database, built in this study, of the most frequently isolated species, including Fusarium solani, Fusarium oxysporum, Fusarium verticilloides, Fusarium proliferatum and Fusarium dimerum, correctly identified 57 strains. As expected, the four species (i.e. Fusarium chlamydosporum, Fusarium equiseti, Fusarium polyphialidicum, Fusarium sacchari) not represented in the database were not identified. Results from mass spectrometry and molecular identification agreed in five of the six cases in which results from morphological and molecular identification were not in agreement. MALDI-TOF yielded results within 1 h, making it a valuable tool for identifying clinical Fusarium isolates at the species level. Uncommon species must now be added to the database. MALDI-TOF may also prove useful for identifying other clinically important moulds.

Successful treatment of disseminated Geotrichum capitatum infection with a combination of caspofungin and voriconazole in an immunocompromised patient.

Disseminated Geotrichum capitatum infection is uncommon, and has been reported exclusively in immunocompromised patients. The prognosis is poor with a mortality rate of approximately 50-75%. We report a case of disseminated G. capitatum infection in a severely neutropenic patient who was receiving chemotherapy for acute myeloblastic leukaemia. G. capitatum was isolated from blood cultures, skin lesions, bronchoalveolar lavage fluid, throat swabs and stools. The infection was successfully cured with a combination of voriconazole and caspofungin.

Production of monoclonal antibodies directed against the microsporidium Enterocytozoon bieneusi.

Several hybridomas producing antibodies detected by indirect immunofluorescence antibody test (IFAT) were established by fusion of mouse myeloma SP2/O with spleen cells from BALB/c mice immunized against whole spores (protocol 1) or chitinase-treated spores (protocol 2) of Enterocytozoon bieneusi and were cloned twice by limiting dilutions. Two monoclonal antibodies (MAbs), 3B82H2 from protocol 1, isotyped as immunoglobulin M (IgM), and 6E52D9 from protocol 2, isotyped as IgG, were expanded in both ascites and culture. IFAT with the MAbs showed that both MAbs reacted exclusively with the walls of the spores of E. bieneusi, strongly staining the surface of mature spores, and produced titers of greater than 4,096. Immunogold electron microscopy confirmed the specific reactivities of both antibodies. No cross-reaction, either with the spores of the other intestinal microsporidium species Encephalitozoon intestinalis or with yeast cells, bacteria, or any other intestinal parasites, was observed. The MAbs were used to identify E. bieneusi spores in fecal specimens from patients suspected of having intestinal microsporidiosis. The IFAT was validated against standard staining methods (Chromotrope 2R and Uvitex 2B) and PCR. We report here the first description and characterization of two MAbs specific for the spore wall of E. bieneusi. These MAbs have great potential for the demonstration and species determination of E. bieneusi, and their application in immunofluorescence identification of E. bieneusi in stool samples could offer a new diagnostic tool for clinical laboratories.

Invasive Fusarium infections: a retrospective survey of 31 cases. The French 'Groupe d'Etudes des Mycoses Opportunistes' GEMO.

A retrospective study was conducted in France to investigate Fusarium infections which are now recognized as emerging opportunistic infections. The clinical and mycological findings for 31 cases diagnosed between 1984 and 1993 by members of the French Groupe d'Etudes des Mycoses Opportunistes were analysed. All suffered from haematological disease, most often acute leucaemia (n = 19). Twenty-two had received cytostatic chemotherapy and ten had undergone bone marrow transplantation. Prolonged aplasia and pancytopenia were present in 18 and 11 patients, respectively. Skin (61%) and blood (42%) were the sites most frequently involved. Fusarium solani (n = 7), Fusarium oxysporum (n = 7), Fusarium verticilloides (n = 7) were the species most frequently isolated. Nine antifungal treatments were used, associated with colony-stimulating factors in five cases. None was unambiguously superior to all the others. The overall mortality was 51.6% with a specific mortality > or = 25.8%. The disseminated form of the infection was associated with poor prognosis (P < 0.02) whereas improving granulocyte count improved prognosis (P < 0.001). More aggressive cytostatic regimens used for patients with haematological malignancies have favoured the emergence of Fusarium infections. As prognosis is closely correlated with neutrophil recovery, the promising results obtained with the use of colony-stimulating factors should be further evaluated.

Transmission of Plasmodium falciparum by allogeneic bone marrow transplantation.

We describe a case of Plasmodium falciparum infection in a Comorian patient undergoing BMT. The patient's last visit to an endemic area was 1 year prior to BMT. The donor left the Comoro Islands 2 months before marrow harvesting. They had both had previous episodes of malaria and were seropositive for Plasmodium falciparum. At the time of BMT, blood smears were negative in both the donor and recipient. On day 12 post-BMT the patient was asymptomatic but a blood smear revealed 12.5% parasitemia. We consider that donors and recipients at risk pre-BMT should routinely be given specific treatment before marrow harvesting and conditioning, independent of the appearance of blood smears.

Association of lipid accumulation in small intestinal mucosa with decreased serum triglyceride and cholesterol levels in AIDS.

Lipid accumulation has been described in the duodenal lamina propria of human immunodeficiency virus (HIV)-infected patients with diarrhea and malabsorption. Using light and electron microscopy, we studied duodenal biopsies obtained from 54 consecutive HIV-infected patients by means of upper gastrointestinal endoscopy after an overnight fast. The presence of diarrhea and weight loss were recorded, and all the patients had standard stool study for ova, parasites, and bacteria. Serum levels of albumin, triglycerides, and cholesterol were obtained within one week of the endoscopy. Fecal fat and fecal alpha 1-antitrypsin clearance were measured in 11 patients. Lipid accumulation was observed in nine patients (16.6%). Fat droplets were seen in enterocytes, in their basolateral membrane spaces, and in the lamina propria. The mean serum levels of triglycerides (1.85 +/- 0.20 mmol/liter) and cholesterol (2.81 +/- 0.30 mmol/liter) were significantly lower in the patients with enteric steatosis than in patients without this anomaly (respectively, 3.38 +/- 0.39 and 3.97 +/- 0.18 mmol/liter, P < 0.001 P < 0.01). The mean amount of fecal fat in the three patients with lipid accumulation (16 +/- 1.60 g/24 hr) was significantly larger than in the eight patients without lipid accumulation (4.50 +/- 0.62 g/24 hr, P < 0.01). These findings suggest that fat malabsorption in HIV-infected individuals is due to a blockage of transport through the duodenal mucosa. The frequency of diarrhea, weight loss, or identified enteric pathogens did not differ significantly between patients with and without enteric steatosis. Both the etiology and the pathophysiology of these alterations remain to be documented.

[Subacute Plasmodium falciparum malaria in 43 patients returning from areas with chloroquine in Africa]. / Paludisme subaigu à Plasmodium falciparum chez 43 sujets revenant de régions de chloroquinorésistance en Afrique.

PURPOSE: To identify clinical and biological features of subacute falciparum malaria, risk factors, and to evaluate the efficacy of curative treatment. PATIENTS AND METHODS: Diagnostic criteria were the association of apyrexia, anemia, little or no parasitemia and a high titer of anti-Plasmodium antibodies. Forty-three cases were observed in subjects returning from chloroquine-resistant areas in Africa. They were matched with controls for age, country of residence and duration of stay. Controls were missionaries who attended our unit for a routine medical check-up during the study period. RESULTS: The clinical presentation and biological features were similar to "malarial cachexia", a condition mainly described in non-immune children in endemic areas. Splenomegaly was present in 58% of the patients. Biological features included little or no parasitemia, an overall decrease in the blood cell count, an increased erythrocyte sedimentation rate and a high titer of anti-Plasmodium antibodies. This syndrome was not correlated with the frequency of chloroquine resistance, the area of stay (urban or rural) or to the kind of chemoprophylaxis. CONCLUSIONS: This study describes subacute resistant falciparum malaria in patients who had prolonged stay in chloroquine-resistant areas of Africa associating splenomegaly, cytopenia and a low or absent parasitemia. Subacute chloroquine-resistant malaria could be due to host factors which remained to be determined by prospective immunological studies. Curative treatment with mefloquine is effective.

Disseminated Penicillium marneffei infection associated with human immunodeficiency virus: a report of two cases and a review of 35 published cases.

Penicillium marneffei, a dimorphic fungus that is endemic in southeast Asia, causes deep-seated infection in humans and rodents. About 20 cases have been reported among the local populations of China, Thailand, and Hong Kong, and 35 cases have now been described in patients infected with the human immunodeficiency virus (HIV). We present a review of the literature and report two additional cases. Both immunocompromised and apparently immunocompetent hosts tend to develop disseminated, symptomatic infection. HIV-infected patients having travelled to southeast Asia and presenting with fever, skin lesions, hepatomegaly, adenopathies, or lung disease should be investigated for Penicillium marneffei infection. The diagnosis is based on the demonstration of the organism in clinical specimens. Treatment with amphotericin B or itraconazole is generally successful, but maintenance therapy is warranted for patients with an underlying immunodeficiency.

Toxoplasma gondii pneumonia in patients with the acquired immunodeficiency syndrome.

PURPOSE: To perform a retrospective and descriptive study of Toxoplasma gondii pneumonia in patients infected with the human immunodeficiency virus (HIV). Clinical presentation, diagnostic procedures, results of therapy, and hypotheses on pathophysiology are discussed. PATIENTS AND METHODS: The study consisted of 13 HIV-infected patients who had developed T. gondii pneumonia. Eight had acquired immunodeficiency syndrome (AIDS) prior to T. gondii pneumonia and three of them had non-Hodgkin's lymphoma. Mean CD4 cell count was 32 x 10(6)/L. Serum anti-toxoplasma antibody titers were measured by an indirect hemagglutination assay and/or by an indirect immunofluorescence assay. RESULTS: All patients had fever and bilateral pulmonary infiltrates; two of them presented with septic shock. Mean arterial oxygen tension was 47 +/- 12 mm Hg. The diagnosis was established by bronchoalveolar lavage in 10 of 11 cases, open lung biopsy in one case, and postmortem biopsy in two cases. Serologic evidence of past infection was observed in 11 of 12 cases, while one patient presented with acute disseminated disease and absence of serum anti-toxoplasma antibody response. Extrapulmonary involvement was present in seven patients: liver (four), brain (three), bone marrow (two), heart (two), stomach (one). Ten patients recovered from T. gondii pneumonia. CONCLUSION: T. gondii pneumonia must be considered in AIDS patients with severe diffuse bilateral pneumonia, especially when associated with a very low CD4 cell count or non-Hodgkin's lymphoma. In most of these cases, disseminated disease was associated with reactivation of prior latent infection.

[Needle aspiration biopsy in the diagnosis of cutaneous leishmaniasis]. / Prélèvement par aspiration à l'aiguille dans le diagnostic de la leishmaniose cutanée.

The classic diagnostic procedure for cutaneous leishmaniasis is based on the examination of Giemsa-stained smears made from the fluid obtained by scraping the edges of the lesion with a lancet, or prepared with small plugs of superficial tissues. In this report, we compare the results given by this standard method with those obtained by the examination of needle aspirates. Aspirates are secured by injecting a few millimetres outside the external border of the lesion 0.3 to 0.5 ml of saline through a thin needle, rubbing the injured skin, and thereafter pulling back slowly the plunger. Amastigotes were found in all smears from needle aspirates, and in only 11 out 15 obtained by scraping. Aspirates were also more suitable for cultures of parasites.

[Value of cytapheresis in the treatment of loaiasis with high blood microfilaria levels. Results in 7 cases]. / Intérêt de la cytaphérèse dans le traitement des loases à fortes microfilarémies. Résultats à propos de sept cas.

Seven patients infected with the filarial worm Loa loa received a treatment by cytapheresis in an attempt to lower the microfilaraemia. Microfilarial levels of between 6,000 and 38,500 ml, before extraction, were reduced, according to the case, by between 47 and 97% (mean 76%). The diethylcarbamazine chemotherapy which followed in 6 of 7 patients showed no sign of any of the serious side-effects which often occur in these type of cases. Due to its practicality and the fact that it is well tolerated, both clinically and biologically, cytapheresis would seem to represent the best method for initially treating loaiasis with high microfilaremia.

Systemic infections with Trichosporon beigelii (cutaneum). Report of three new cases.

Three new cases of systemic mycosis due to Trichosporon cutaneum are reported and compared with the 23 previous reports. Two patients had acute leukemia and one patient had a lymphoblastic lymphoma. Blood cultures in two patients and cerebrospinal fluid in the third patient were positive for T. cutaneum. Only one patient recovered after antimycotic therapy and concomitant remission of his leukemia. At autopsy, the two other patients showed widespread infection with T. cutaneum. The authors conclude that diagnosis and management of such infection in the immunosuppressed host are difficult and the prognosis is poor.

[Effect of the use of drinking water on the pathogenic fecal flora in a Sahel rural region]. / Retentissement sur la flore fécale pathogène de l'utilisation d'une eau potable en milieu rural sahélien.

Within the scope of a village hydraulic programme, in a Sahel rural area, an investigation was conducted in a school to determine the effect of the use of drinking water on fecal flora. One year following the installation of a hydraulic pump, it was observed that pathogenic salmonellae, shigellae and amoebic cysts had virtually disappeared. In contrast, the perennial character of viral contamination was demonstrated. Water guaranteed potable, therefore, appears to be an essential factor in the prevention of infections due to fecal pathogenic microorganisms. It is essential, however, that the areas surrounding the pumps be subjected to strict supervision to avoid pollution of the underground water sheet.

Hepatocytes as feeder-layers for in vitro cultivation of Plasmodium falciparum blood-stages.

To improve the in vitro growth of Plasmodium falciparum we attempted to cultivate its erythrocytic stages on monolayers of functionally active hepatocytes. Hepatocytes from Swiss Albino mice were isolated by perfusing the liver with a collagenase solution and were co-cultured with a liver epithelial cell type in RPMI 1640 medium supplemented with 10% human umbilical cord serum. The results show that the presence of hepatocytes improves both the multiplication rates of three strains of P. falciparum already in cultivation and the proliferation of freshly isolated strains. Of nine primary isolates tested, only three could be adapted in the standard conditions, whereas all grew readily in the presence of hepatocytes. After two to three weeks of culture with feeder cells, all the strains could be maintained continuously in standard conditions. Similar results were obtained using hepatocytes from another rodent species. Growth was also improved using the supernatant from hepatocyte cultures. No improvement resulted from the use of two human hepatoma cell lines, one rat hepatoma, human embryonic lung fibroblasts, human liver fibroblasts and rat liver epithelial cells as feeder layers. From these results it appears that better culture media can be designed and that the effect of hepatocytes is probably related to the specific functions exhibited by these cells. Hepatocytes may act either by removing toxic substances, particularly lactic acid in the Krebs and Cori cycles, and/or supplying nutrients essential to the parasite.

[Treatment of urinary and intestinal bilharziosis with praziquantel. Apropos of 182 cases]. / Traitement de la bilharziose urinaire et intestinale par praziquantel. A propos de 182 observations.

182 patients representing 188 cases of schistosomiasis (including 152 S. haematobium, 34 S. mansoni, and 2 S. intercalatum cases) were treated with praziquantel using 40 mg/kg in a single intake. Biological check-up did not reveal any abnormality that could have been related to the treatment. Clinical side-effects appeared in 25% cases and were essentially gastrointestinal symptoms, headaches, dizziness. Efficiency was assessed by direct parasitological examinations of the feces or urine that were performed after treatment. It reached 91% for S. haematobium and 95% for S. mansoni schistosomiasis.