[Neuromeningeal cryptococcosis in Mali]. / La cryptococcose neuro-méningée au Mali.

Cryptococcal meningitis is the most common fatal central nervous system infection in AIDS patients in Sub-Saharan Africa. The purpose of this prospective study conducted from March 2003 to February 2004 in the internal medicine and infectious diseases departments of the Point G University Hospital Center was to investigate the clinical, prognostic and epidemiological profile of Cryptococcus neoformans infection in patients hospitalized for brain and meningeale infection (BMI). Diagnosis of neuromeningeal cryptococcosis (NMC) was based on positive identification of Cryptococcus by direct exam of the cebrospinal fluid (CSF) after India ink staining and/or culture on Sabouraud medium without actidione. During the study period, a total of 569 patients were hospitalized including 235 (41.3%) with HIV infection. Overall C. neoformans was identified in 14 patients. Median patient age was 39 +/- 8 years. There was a male preponderance with a sex ratio of 1.8 (9 men/5 women). Patients with BMI were HIV-positive in 85.7% of cases (n=12) and HIV-negative in 14.3% (n=2). The overall and HIV-specific prevalence of BMI was 2.5% and 5.1% respectively. The CD4 lymphocyte count was between I and 49 cells/mm3 in 64.3% of cases. The main clinical symptoms were cephalea in 85.7% of cases, altered consciousness in 50% and nausea/vomiting in 35.7%. Neurological manifestations (hemiparesis and cranial nerve deficit) were noted in 14.3%. HIV infection is the main purveyor of NMC in Mali. The actual incidence of cryptococcosis is unclear due to the poor sensitivity of diagnostic techniques. This study highlights diagnostic difficulties related to clinical polymorphism and poor technical facilities. Agglutination testing of blood and CSF is recommended, but mortality remains.

[Travelers' knowledge of vaccines. A survey including 400 people having consulted an international vaccination center]. / Connaissances des voyageurs à propos des vaccins. Enquête auprès de 400 personnes consultantes d'un centre de vaccinations internationales.

UNLABELLED: The authors conducted a survey in an international vaccination centre. The aim of the study was to assess the travelers' knowledge of vaccines and to check their health record to determine the vaccination rate. We used a self-administered questionnaire and added vaccination certificates to the document. RESULTS: Most of the travelers knew about complications due to vaccinations, but ignored their contraindications. Knowledge decreased over the age of thirty. The travelers agreed with antihepatitis B vaccination and only 10% mentioned the risk of multiple sclerosis. The list of compulsory vaccinations and their schedule were well-known but some of the travelers ignored their vaccine status and said they trusted their general practitioner. Only 40% of people over 20years of age were able to present their national or international vaccination certificate, a reason for underestimating the vaccination rate. CONCLUSION: It would be necessary to have a "World Health Organization" type vaccination certificate available for all French people over 20years of age, while improving the smart health card and training practitioners to better inform their patients about vaccinations.

[Neuromeningeal cryptococcosis in non-HIV patients to CHU ward of Point G in Bamako (Mali): 3 case report]. / Cryptococcose neuroméningée chez des patients VIH négatif au CHU du Point G de Bamako (Mali): à propos de trois observations cliniques.

We report 3 clinical observations of neuromeningeal cryptococcosis which occurred without any immunodepression related to HIV infection. Our patients were male. They did not present any particular medical history indicating a diagnosis of cryptococcosis. Nevertheless we found a professional exposure to the risk of inhalation of C. neoformans capsules. The diagnosis was based on presence of encapsulated yeast of Cryptococcus in the direct exam by China ink and culture on Sabouraud medium. CD4 lymphocytic count was done in two patients (case 1:899 cells/mm3; case 2:347 cells/mm3). Idiopathic lymphocytopenia was noted in one case. Co-morbidity of 5. pneumoniae meningitis was reported in one patient (case 3). Treatment was based on injectable amphotericin B in monotherapy (case 1), followed by perfusion of fluconazole cure (case 2). Case 3 was treated by perfusion of amphotericin B associated with ceftriaxone (case 2). No secondary prophylactic treatment was administered. Evolution was favorable after follow-up of 5 years (case 1) and of 4 years (case 2). The third patient died during hospitalization.

[Breast abscess with Salmonella typhi and review of the literature]. / Un abcès du sein à Salmonella typhi et revue de la littérature.

We report the case of a 54-year-old woman who presented with breast abscess, which appeared through a common alimentary toxi-infection with Salmonella Typhi, infection, which implied twelve patients having attended the same restaurant. With around hundred native cases a year in France, typhoid fever is not a very frequent toxi-infection. Among the known extra-intestinal manifestations of Salmonella infections, the breast abscess remains rare and the literature revealed less than ten published cases, including some revealed the disease. In our observation, the imputability of S. Typhi was retained based on the chronology of the clinical signs, specific treatments, and the successful outcome under antibiotherapy, in spite of the negativity of the breast abscess bacteriological samples. We also analyze rare cases of breast abscess due to S. Typhi found in the literature.

Management of an Acinetobacter baumannii outbreak in an intensive care unit.

INTRODUCTION: Acinetobacter baumannii is a ubiquitous pathogen resistant to desiccation and responsible for healthcare-associated infections (HAI), especially in intensive care units (ICU) where it is responsible for 5-10% of HAIs. An A. baumannii outbreak occurred in the ICU of the University Hospital of Angers, France. OBJECTIVES: To describe the A. baumannii outbreak and to evaluate the control measures taken. The secondary objective was to evaluate the impact of the electronic alert system on the incidence of multidrug resistance to antibiotics. METHODS: We performed a descriptive study of A. baumannii carriers during the outbreak. Case contacts and carriers were described using the epidemic curve and a case synopsis table. RESULTS: From August 2011 to September 2013, 49 patients presenting with an extended-spectrum beta-lactamase-producing A. baumannii infection were identified: thirty-four were colonized and 15 were infected. No death was due to the outbreak. Measures taken were: geographical and technical isolation of patients, dedicated team implementation, contact precaution implementation including hand hygiene measures, appropriate use of gloves, and reinforcement of bio-cleaning procedures. CONCLUSION: Some patients were re-admitted to hospital while still being carriers; this could explain epidemic peaks. The immersion mission of the hygiene nurse contributed to answering healthcare workers' queries and led to a better cooperation between the ICU and the hygiene team.

Tetanus associated with road accidents in the infectious diseases department of Point G University Hospital, Bamako, Mali.

The aim of this study was to describe the epidemiological, clinical, and prognostic aspects of tetanus associated with road accidents and to make recommendations. This observational study collected retrospective clinical data over a 9-year period about adults admitted for trismus and/or generalized or localized paroxysm after a road accident. The study included 25 patients, accounting for 22.12 % of all tetanus cases. Men were massively overrepresented (sex-ratio M/F: 24/1). The median age was 34 ± 8 years. In all, vaccination status was unknown for 4 patients and known to be negative for 21. Immunoprophylaxis was nonexistent in all cases. The generalized clinical form was dominant (96 %). Severity reached level III for 12 % of patients. The points of entry included open leg fractures (4 cases), head wounds (2), mucocutaneous wounds (14), and muscle contusions (5). The mean time to referral for tetanus was 8 ± 7 days, and the median hospital stay 9.08 ± 11 days. Patients were mostly residents of urban (56 %) and suburban areas (28 %) [P = 0.04]. Two cases were complicated by severe malaria. The mortality rate was 60 %, and 52 % of the deaths occurred within the first 72 hours after hospitalization. It is essential to promote serum therapy and tetanus immunization for patients after road accidents. Increasing the awareness of traditional healers of these treatments deserves consideration.

[A new case of bat rabies in France]. / Un nouveau cas de rage chez les chauves-souris en France.

INTRODUCTION: Bat rabies represents an emerging zoonosis in Europe and the only endemic cause of rabies in France. CASE RECORD: A 29 year-old woman was bitten at the hand by a bat. The diagnosis of bat rabies was positive and the viral strain was an European Bat Lyssavirus 1a. A combination of rabies vaccine and human rabies immune globulin was provided to the patient. DISCUSSION: Any direct contact with a bat must be avoided. In case of exposure to bats, the postexposure treatment must associate the rabies vaccine and human rabies immune globulin because of antigenic diversity of Lyssavirus circulating in bat species.

Appearance of host-specific nucleolar proteins in intranuclear "dense bodies" following herpes simplex infection.

Nucleolar modifications induced by herpes simplex virus type 1 (HSV1) infection were studied at the ultrastructural level with special attention to the fate of a family of proteins serologically related to the nucleolar 100 kDa protein. Immunocytochemical techniques revealed that antigenic sites related to these proteins were associated with nucleoli in a pattern similar to that observed with non-infected cells. In addition, the "dense bodies" induced by HSV infection were heavily decorated by antibodies to the 100 kDa protein. Neither DNA nor RNA was detectable in the latter by cytochemical techniques. Therefore, it appears that "dense bodies" are exclusively proteinaceous and contain at least one host protein implicated in ribosomal RNA synthesis. An accumulation of 100 kDa protein in extra-nucleolar structures might account for previously reported defects in ribosomal RNA expression during HSV infection.

Cytochemical study of the localization and organization of parental herpes simplex virus type I DNA during initial infection of the cell.

Changes in the location and structural organization of parental herpes simplex virus type 1 (HSV-1) DNA during its migration from the extracellular space to the interior of the nucleus of the target cell were examined by in situ hybridization using an HSV-1 DNA probe, specific DNA staining, and autoradiography after infection of cells with tritium-labeled viruses. In situ hybridization was carried out on denatured DNA to reveal as much as possible of the HSV-1 sequence present at the surface of the sections, and also on non-denatured DNA which revealed the presence of single-stranded portions of parental DNA, both prior to and during its intracellular migration. The results from in situ hybridization and autoradiography demonstrated that a short interval of about 15 min separated the initial contact of the viruses with the cells from the entry of parental viral DNA into the nucleus. In transit, morphologically intact nucleoids were released into the cytoplasm, and swollen nucleoids which contained partially decondensed viral DNA became detectable in the juxtanuclear cytoplasm and the periphery of the nucleus among the cell chromatin fibers. Completely decondensed parental viral DNA fibers could not be distinguished structurally from cellular DNA, but their position could be revealed by the in situ hybridization label. The infective DNA became randomly distributed within all compartments of the nucleus except the matrix-associated clusters of interchromatin granules.

Alterations of nucleolar ultrastructure and ribosome biogenesis by actinomycin D. Implications for U3 snRNP function.

We have studied, at the electron microscope level, the reorganizations of nucleolar ultrastructure induced by actinomycin D (AMD) in different conditions of drug treatment associated with an inhibition of rRNA synthesis. We have analyzed in parallel the localizations of ribosomal genes, of their transcripts, of various pre-rRNA intermediates, as well as of U3 RNA and fibrillarin by in situ hybridization with nucleic acid probes and immunocytological detection on thin sections of human and mouse cells. Consistent with previous observations, dense fibrillar component (DFC) and granular component (GC) appear to contain distinct pre-rRNA species at different stages of their processing. DFC appears as a major site of U3 RNA accumulation, but a very substantial fraction of nucleolar U3 RNA is also found in GC, colocalizing with partially processed pre-rRNAs. Remarkably, the major nucleolar components retain their ultrastructural appearance when extensively depleted of their pre-rRNA moiety, and ribosomal genes are always detected over fibrillar center (FC), even after extended AMD treatments which result in the characteristic segregation of nucleolar components. Moreover, while for GC the U3 RNA and pre-rRNA contents evolve in parallel following the cessation of rRNA synthesis, a dramatic uncoupling is observed for DFC. The persistent presence of U3 RNA and fibrillarin after pre-rRNA depletion suggests that DFC could represent an anchorage site for U3 snRNPs, before their entering another cycle of pre-rRNA processing reactions.

Superiority of in situ hybridization over immunolabeling for detecting DNA on Lowicryl sections: a study on adenovirus-infected cells.

We investigated the intranuclear distribution of viral genomes in adenovirus Type 5-infected HeLa cells on ultra-thin sections of Lowicryl K4M-embedded material by immunolabeling of DNA and in situ hybridization with a viral DNA probe. Monoclonal AC-30-10, raised against double- and single-stranded DNA (dsDNA, ssDNA), and HB2 antibodies, raised against only the dsDNA, were used. Both antibodies intensely labeled the condensed host chromatin and the virus-induced substructures containing inactive encapsidated and non-encapsidated viral genomes. The antibodies labeled only slightly those substructures containing replicating and transcribing viral genomes. Viral ssDNA, which is accumulated within well-delineated compact fibrillar structures, was not revealed by AC-30-10 even after elimination of the proteins of the section. In situ hybridization detected all of these types of viral DNA, depending on the protocol used. Therefore, the sensitivity of in situ hybridization for detecting DNA appears superior to immunolabeling with specific antibodies.

Cytochemical analysis of DNA organization during encapsidation in herpes simplex virus.

The organization of intranuclear Herpes simplex virus DNA in rabbit fibroblast cells infected for 7 hr with HSV type 1 was examined before and during encapsidation by electron microscopic cytochemistry. Most non-encapsidated viral deoxyribonucleoprotein fibers exhibited a non-nucleosomal configuration. Empty capsids within the virus-specific regions of infected nuclei were wrapped with portions of the viral genome which adhered tightly to their surfaces even under conditions that loosened and spread apart other nucleoprotein fibers. During encapsidation, the internal surface of the capsid shell also appeared to bind a part of the viral genome, specifically the outer cage portion, which is detectable in methanol-dehydrated cells. Variations in the amount of DNA within the capsids indicated that the insertion of HSV genome into the capsid is a progressive process. The cage and core cylinder portions of the viral nucleoid appear to form and develop simultaneously. We propose that there may be binding sites on both the external and internal surfaces of the capsid shells which might play a role in the encapsidation process.

Effect of dehydrating agents on DNA organization in herpes viruses.

With routine procedures of Epon- or GMA-embedding and a stain specific for DNA, the nucleoid of mature herpes simplex virus-type 1 (HSV-1) assumes the well-known form of a short, compact, hollow cylinder or torus. A new, more complex organization of DNA filaments in encapsidated HSV-1 was found in infected cells after aldehyde fixation, methanol dehydration, and Lowicryl embedment. We have determined that it is the use of methanol as dehydrating agent that permits visualization of this internal structure. The same new spatial organization of DNA can be seen in Epon and GMA sections when methanol dehydration is used. This organization is lost in a methanol-ethanol sequence of dehydration but can be restored in an ethanol-methanol sequence. Dimethylsulfoxide (DMSO) is the only other agent among several reviewed here which resembles methanol in its effect on HSV-1 DNA. Methanol had the same effect on five subfamilies of the herpes group (HSV-1, HSV-2, CCV, CMV, CTHV) but did not alter the nucleoid ultrastructure in simian virus 40 (SV40) and adenovirus type 5 (Ad 5). Therefore, it may sometimes, but not always, provide additional information about the organization of biological structures.

Nitazoxanide in the treatment of cryptosporidial diarrhea and other intestinal parasitic infections associated with acquired immunodeficiency syndrome in tropical Africa.

Eighteen patients hospitalized with intestinal parasitic infections associated with diarrhea and dehydration completed a study of nitazoxanide in the treatment of Cryptosporidium parvum and other intestinal parasitic infections. Seventeen of the 18 patients were positive for human immunodeficiency virus. Twelve patients were diagnosed with clinical Stage 4 acquired immunodeficiency syndrome (AIDS) according to the 1990 World Health Organization proposed clinical classification system and cryptosporidiosis. Nitazoxanide (500 mg tablets) were administered orally, one tablet twice a day for seven consecutive days. Cryptosporidium parvum oocysts were eradicated or reduced by more than 95% in seven of the 12 Stage 4 AIDS patients who completed the study based upon two post-treatment fecal examinations conducted on days 7 and 14 following the initiation of treatment. The elimination or reduction of C. parvum oocysts was associated with a complete resolution of diarrhea in four of the seven patients. The test drug was also effective against cases of Isospora belli, Entamoeba histolytica, Giardia lamblia, Ascaris lumbricoides, Enterobius vermicularis, Hymenolepis nana, and Dicrocoelium dentriticum. Treatment with nitazoxanide was well tolerated by the patients. There were no abnormalities in blood chemistry or hematology data that were considered to be attributable to nitazoxanide therapy. Transient episodes of vomiting were observed in four patients, all with Stage 4 AIDS and cryptosporidiosis, which resolved spontaneously without discontinuation of treatment and were not considered to be related to administration of nitazoxanide.

PIP: The effectiveness of nitazoxanide in the treatment of Cryptosporidium parvum and other intestinal parasitic diseases was assessed in 18 patients hospitalized at Point G. National Hospital in Bamako, Mali, with parasite-related diarrhea, dehydration, and weight loss. 17 of the 18 patients were infected with HIV, and 12 of these had progressed to clinical stage 4 AIDS. 500 mg tablets of nitazoxanide were administered twice a day for 7 days. After completion of treatment, Cryptosporidium parvum oocysts were eradicated or reduced by more than 95% in 7 of the stage 4 AIDS patients; diarrhea was completely resolved in 4 of these patients. Nitazoxanide was also effective against other parasites common in AIDS patients, including Entamoeba histolytica, Giardia lamblia, and Isospora belli. The test drug was well tolerated by all recipients, with no blood chemistry abnormalities.

Human cytomegalovirus DNA kinetics using a novel HCMV DNA quantitative assay in white blood cells of immunocompromised patients under Ganciclovir therapy.

The clearance of human cytomegalovirus (HCMV) was evaluated in infected patients under Ganciclovir (GCV) treatment, using a novel HCMV DNA quantitation assay (HCMV DNA hybrid capture system, Murex Diagnostics). Peripheral white blood cells (WBC) from whole blood specimens of seven AIDS patients, three kidney and two allogeneic bone marrow transplant (BMT) recipients suffering from HCMV disease, were assessed by this method. HCMV DNA 50 and 90% mean clearances were observed at 2.11 +/- 1.97 and 6.22 +/- 4.31 days, respectively, after initial GCV treatment. The viral DNA kinetics were correlated with positive and negative pp65 antigenaemia and viral blood culture. Two-fold higher clearances and initial DNA levels were observed in the AIDS group compared to the transplant group. Neither clinical nor virological relapses were observed under GCV treatment. HCMV DNA quantitation in WBC appears well adapted for a therapeutic follow up of patients with HCMV disease.

Specificity of the solid phase alkaline phosphatase immunocapture assay for the diagnosis of human Schistosoma mansoni infection.

The species specificity of the solid phase alkaline phosphatase immunocapture assay (APIA) for the immunological detection of human immunoglobulin G antibodies to the alkaline phosphatase of adult Schistosoma mansoni was evaluated. Sera from schistosomiasis patients from South America, West Africa, south-east Asia and uninfected control subjects were compared. Only the sera of patients infected with S. mansoni gave positive results. There was no apparent difference between 2 populations infected with S. mansoni, one from South America and the other from West Africa. The results with sera from various regions of West Africa were also indistinguishable. Although the APIA was not able to discriminate the geographical origin of the S. mansoni-infected subjects, the method appeared to be specific for S. mansoni and suitable for use in the immunodiagnosis of schistosomiasis mansoni, particularly in endemic areas where mixed infections of Schistosoma spp. occur.

Synthesis and maturation of viral transcripts in herpes simplex virus type 1 infected HeLa cells: the role of interchromatin granules.

The response of the cellular RNA processing machinery to herpes simplex virus type 1 (HSV-1) infection was studied at the ultrastructural level in HeLa cells and compared to the distribution of RNA polymerase II molecules and viral RNA. Immunogold labeling of RNA polymerase II molecules revealed that viral genome transcription was restricted to filaments in an intranuclear, virus-induced region. This region also contained viral RNAs as revealed by in situ hybridization of two biotinylated viral DNA probes: a probe encompassing a limited portion of the viral genome (the F fragment) and a probe for the total genome. In addition, the latter probe revealed large amounts of viral RNA within the clusters of interchromatin granules, intranuclear structures of normal cells that became enlarged during HSV-1 infection. Components of spliceosomes were localized by in situ hybridization with biotinylated U1 and U2 DNA probes. The large viral region contained only traces of U1 and U2 RNAs, probably because of the low frequency of splices of viral transcripts. The clusters of interchromatin granules, however, accumulated U1 and U2 RNAs with the same frequency as in noninfected cells. Poly(A) RNA was detected by in situ hybridization of a biotinylated poly(dT) probe. Some was present over the filaments of the virus-induced region but most was accumulated in the clusters of interchromatin granules. Our data suggest, therefore, that the clusters of interchromatin granules, in addition to their involvement in spliceosome component assembly, might also be a transient storage site for some families of viral mRNA, possibly a sorting site that regulates their migration.

[Efficacy of diclofenac continuous perfusion in hyperalgic bone metastases. Apropos of 20 cases treated at Gustave Roussy Institute]. / Efficacité du diclofenac en perfusion continue dans les métastases osseuses hyperalgiques. A propos de 20 cas traités à l'Institut Gustave-Roussy.

In this trial, 20 patients suffering from bone metastasis pain received diclofenac, a non-steroid anti-inflammatory agent, by continuous IV infusion. No pain was observed in 77% after 2.7 days of treatment. 33% of bedridden patients were able to move again. Two patients (10%) presented gastric pain controlled by symptomatic treatment.