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Biomedical subjects

R Negroni

Publications and source records attributed to R Negroni.

At least 19 recordsLinked to original sources

Biogeographic range expansion into South America by Coccidioides immitis mirrors New World patterns of human migration.

Long-distance population dispersal leaves its characteristic signature in genomes, namely, reduced diversity and increased linkage between genetic markers. This signature enables historical patterns of range expansion to be traced. Herein, we use microsatellite loci from the human pathogen Coccidioides immitis to show that genetic diversity in this fungus is geographically partitioned throughout North America. In contrast, analyses of South American C. immitis show that this population is genetically depauperate and was founded from a single North American population centered in Texas. Variances of allele distributions show that South American C. immitis have undergone rapid population growth, consistent with an epidemic increase in postcolonization population size. Herein, we estimate the introduction into South America to have occurred within the last 9,000-140,000 years. This range increase parallels that of Homo sapiens. Because of known associations between Amerindians and this fungus, we suggest that the colonization of South America by C. immitis represents a relatively recent and rapid codispersal of a host and its pathogen.

Base Sequence↗

High serum interleukin-10 and tumor necrosis factor alpha levels in chronic paracoccidioidomycosis.

In patients with chronic paracoccidioidomycosis (n = 10), levels of tumor necrosis factor alpha, interleukin-10, and interleukin-2 in serum, measured by enzyme-linked immunosorbent assay (in picograms per milliliter, as mean +/- standard error of the mean), were higher than in normal controls (n = 8): 186 +/- 40 versus 40 +/- 7 (P < 0.05), 203 +/- 95 versus 20 +/- 8 (P = 0.001), and 96.3 +/- 78.57 versus 1.19 +/- 1.19 (P = 0.045), respectively. Gamma interferon and interleukin-4 levels were similar in patients and controls.

Adult↗

[Tibial abscess caused by histoplasma capsulatum].

Disseminated histoplasmosis is the most serious form of the disease produced by the fungus Histoplasma capsulatum. Histoplasmosis was first described by Darling in 1906, and it is endemic in certain areas of Central and Southeastern regions of USA, and most Latin America countries, including the Pampa húmeda and Mesopotamia regions in Argentina, but in other continents it has a limited regional distribution. Lung involvement predominates in 90% of cases, but H. capsulatum may involve bone and soft tissues. Bone lesions without other signs of infection are very rare, and are often misdiagnosed as cancer. We report a case of disseminated histoplasmosis in a man with a myelodysplasia who presented a left tibial abscess, without any clinical evidence of pulmonary involvement. The patient was successfully treated with itraconazole.

Abscess↗

[Improved prognosis in HIV/AIDS related multi-drug resistant tuberculosis patients treated with highly active antiretroviral therapy].

A prospective cohort study was carried out in patients assisted in the F. J. Muñiz Infectious Disease Hospital, with the aim of determining the effectiveness of highly active antiretroviral therapy (HAART) implemented as soon as the sputum smear microscopy became negative (1 to 3 months) in the survival improvement of HIV/AIDS related multidrug-resistant tuberculosis patients. The cohort was recruited from June 1997 to February 1999 and compared with a pre-HAART control group that consisted of 43 patients. The follow-up of the patients was terminated June 2000. A total of 48 patients who received HAART precociously were included. The mortality rate in this group was 31.2% and the survival time of deceased patients 15.8 +/- 8.5 months. The T lymphocytes CD4+ count was initially 40.1 +/- 30.2/microL, while at the end of the observation period it was 140.4 +/- 73.04/microL and 79.1% of these patients presented undetectable viral load. In the control group the overall mortality was 90.7% and the survival time of deceased patients 8.95 +/- 3.72 months. We conclude that the early anti-retroviral therapy, together with the treatment of the multidrug-resistant tuberculosis and of other AIDS associated diseases represent a useful approach to achieve a longer and better survival in these severely immunodepressed patients.

AIDS-Related Opportunistic Infections↗

[Evaluation of a new chromogenic medium (Candida ID) for the isolation and presumptive identification of Candida albicans and other medically important yeasts].

Candidiasis is a frequent human infection caused mainly by Candida albicans. However, other species are emerging as important pathogens, as Candida glabrata, Candida parapsilosis, Candida tropicalis, Candida krusei or Candida guilliermondii. Rapid identification of clinical isolates could facilitate diagnosis and treatment. Candida ID (bioMerieux, Spain) is a new medium for the isolation and presumptive identification of yeasts: C. albicans grows as blue colonies, and C. tropicalis, C. guilliermondii, Candida kefyr and Candida lusitaniae as pink ones. The utility of Candida ID was evaluated with more than 700 clinical isolates and type culture collection strains from different genera including Candida, Cryptococcus, Saccharomyces, and Rhodotorula. Presumptive identification was confirmed by germ tube test, microscopic morphology and chlamydoconidia production on corn meal agar and carbohydrate assimilation on API-ATB ID 32C or Vitek (bioMerieux). Growth on Candida ID was rapid (18-24 h) for most of the yeast strains tested. Sensitivity and specificity of identification of C. albicans was significantly high (>98%), since a very low number of isolates were found to be false negative or false positive. A better result was obtained for species growing as pink colonies (>99.5%). Detection of different species of medical important yeasts was easy with Candida ID, as perfectly distinct colors and textures of colonies were observed on this medium. Candida ID allowed the discrimination between C. glabrata (creamy and smooth) and C. krusei (rough and white) colonies. Other species showed different colony textures and colours, white being the predominant colour. Candida ID was very useful for the presumptive identification C. albicans isolates.

English Abstract↗

Experimental coccidioidomycosis in hamsters. Disease kinetics and death curve in relation to infective dose.

A study of experimental coccidioidomycosis in hamsters (Mesocricetus auratus) is presented. Two experiments were conducted on 75 animals inoculated intracardially with the mycelial form of Coccidioides immitis. The first research (experiment I) studied the kinetics of experimental disease in 15 hamsters inoculated with 300 C. immitis arthroconidia. The parameters studied were: (a) presence of macroscopic lesions in the brain, lungs, liver, spleen and kidneys; (b) microscopic identification of spherules in wet mount preparations of these specimens; (c) samples from all organs cultured at 37 degrees C on Sabouraud glucose agar; (d) blood cultures drawn every 24 h during the first week and subsequently every 48 h and (e) histopathological studies of all organs. The second experiment (experiment II) determined the relationship between the inoculum size and death curve in six groups of 10 animals each, which had received doses of 10, 50, 100, 150, 200 and 300 arthroconidia, respectively. On day 14 post-inoculation, all the animals underwent skin tests and 1 ml of blood was obtained by cardiac puncture to detect antibodies. Disseminated disease with persistent fungaemia developed in all the studied animals. Coccidioides immitis was recovered from all organs, with the lungs being the first to present disease. Death occurred in all groups, regardless of the dose of arthroconidia and 83.3% died between day 22 and day 28 post-infection. The use of this model is proposed for the biological standardization of antigens, the study of prophylactic measures and the "in vivo" evaluation of new antifungal treatments.

Animals↗

The usefulness of blood culture in diagnosing HIV-related systemic mycoses: evaluation of a manual lysis centrifugation method.

The results of 5034 blood cultures, implementing a lysis-centrifugation method with saponin, are summarized in this paper. Three hundred and twenty-two blood samples (6.3%) obtained from a pool of human immunodeficiency virus (HIV)-positive patients yielded fungi. Cryptococcus neoformans was isolated in 199 samples (3.95%), Histoplasma capsulatum in 95 (1.89%). Candida parapsilosis in 12 (0.23%), C. albicans in 7 (0.13%), C. tropicalis in 2, C. krusei in 1, C. guillermondii in 1, and Prototheca wickerhamii in 4 (0.07%). Blood cultures were positive for C. neoformans in 76.23% of patients having a diagnosis of cryptococcosis and in 89.65% of those who had histoplasmosis. The blood culture was the first means of confirming the diagnosis in 23.8% of the patients with cryptococcosis and in 54% with histoplasmosis. In the four patients in whom P. wickerhamii was isolated, a diagnosis of disseminated protothecosis was not achieved by other findings. Catheter infections were responsible for the majority of recovered Candida spp.

AIDS-Related Opportunistic Infections↗

A new PCR primer for the identification of Paracoccidioides brasiliensis based on rRNA sequences coding the internal transcribed spacers (ITS) and 5 x 8S regions.

Internal transcribed spacer (ITS) genes including the 5.8S ribosomal (r)RNA of Paracoccidioides brasiliensis were amplified and the DNA sequences were determined. Based on a comparison of the sequence information, a new polymerase chain reaction (PCR) primer pair was designed for specific amplification of DNA for P. brasiliensis. This primer pair amplified a 418-bp DNA sequence and was 100% successful in identifying 29 strains of P. brasiliensis (including the reference strains) isolated from the regions of Brazil, Costa Rica, Japan, Argentina or from different sources. The results of specificity tests of these primers to compare the fungus with those of Aspergillus fumigatus, Blastomyces dermatitidis, Candida albicans, Cryptococcus neoformans, Histoplasma capsulatum and Penicillium marneffei are also reported.

DNA Primers↗

Disseminated histoplasmosis and AIDS: clinical aspects and diagnostic methods for early detection.

Disseminated histoplasmosis in AIDS patients is the focus of this paper. Cutaneous lesions are reported as a frequent clinical sign. Bone marrow aspiration and biopsy, blood cultures (lysis-centrifugation technique), bronchoalveolar lavage, and skin lesion scrapings are the most effective diagnostic methods. The identification of a specific antigen in blood and urine may be a rapid means of evaluation and follow-up of patients with this disease.

AIDS-Related Opportunistic Infections↗

New aspects of some endemic mycoses.

The treatment of mycetomas varies according to their etiological agents and the clinical state of the patient. For the treatment of eumycetomas, the azole derivatives are the drugs of choice, with itraconazole rendering better results than ketoconazole and presenting better tolerance. Actinomycetomas are treated according to different therapeutic schemes: dapsone plus sulfamethoxazol-trimethoprim (SMT), and streptomycin or amikacin or amoxicillin plus clavulanic acid. The first therapeutic scheme is very useful in the treatment of Nocardia mycetoma, while the association of amikacin plus SMT is the best treatment for those cases produced by Actinomadura madurae. Ciprofloxacin is a very useful drug for the treatment of actinomycotic mycetomas with bone lesions. Although there are several criteria for evaluating clinical outcome there is no accepted criterion of cure. During the 1990s, there was a remarkable increase in the incidence of coccidioidomycosis in California, USA. An almost ten-fold increase in the number of cases was registered during 1992 and 1993 over the usual incidence. A gradual reduction in coccidioidomycosis cases was observed in the late 1990s. This particular coccidioidomycosis outbreak took place in areas of low endemicity, as well as in those of usual high endemicity. Among the factors believed to have influenced this phenomenon were a drought followed by abundant winter/spring rainfall, increased immigration of susceptible individuals, increase in excavation/construction work and a better diagnosis of the infection, particularly in the last part of the decade. The majority of patients presented the usual clinical manifestations of symptomatic primary infection but an unusual number of cases with acute respiratory failure were observed.

Antifungal Agents↗

Mycoses associated with AIDS in the Third World.

Despite advances in diagnosis and treatment, the epidemiological status of the human immunodeficiency virus (HIV) infection is far from under control in most of the developing world. Sub-Saharan Africa, Southeast Asia and India show increased rates of new infections. In Latin America and the Caribbean there were 1.6 million estimated cases of HIV-infected patients at the end of 1997. Fungal diseases have been one of the most relevant diagnoses in relation to the acquired immunodeficiency syndrome (AIDS). Infections due to Candida species and Cryptococcus neoformans var. neoformans are common worldwide. Histoplasma capsulatum, Coccidioides immitis and Penicillium marneffei are important causes of disease in endemic areas. Infection due to Sporothrix schenckii, Blastomyces dermatitidis and Paracoccidioides brasiliensis are uncommon even where they are endemic. Phaeohyphomycetes, hyalohyphomycetes and zygomycetes are still rare as a cause of disease among AIDS patients. However, agents pertaining to these groups, such as Aspergillus spp., have an increasing incidence. Superficial mycoses due to dermatophytes have special features from epidemiological, clinical and therapeutic points of view.

AIDS-Related Opportunistic Infections↗

Histopathological evolution of experimental paracoccidioidomycosis in Wistar rats.

Twenty Wistar rats were inoculated, by the intracardiac route, with 0.5 ml each of a yeast phase suspension of Paracoccidioides brasiliensis Utero strain. The rats were sacrificed at regular intervals post-infection, at which time their lungs, heart, liver, spleen and kidneys were removed, fixed and stained for study. The parameters of interest for the lung specimens were: (a) extent of the lesions; (b) number of fungi; (c) presence of a lymphomononuclear halo. Extrapulmonary lesions were also sought. Until the fourth month post-infection, the lesions were progressive in nature, contained great numbers of viable fungi, and were surrounded by an important lymphomononuclear halo which tended to be confluent. At four and a half months p.i., the extent of the pulmonary lesions was reduced, the granulomas were less compact with fewer viable fungi, macrophages showed microvacuolation, and the lymphomononuclear halo was less pronounced. Extrapulmonary lesions, which were frequently identified in the first months post-infection, diminished from the seventh month onwards. The histological characteristics of extrapulmonary lesions were always the same as those found in the lungs. Infection tended to be controlled by the animals from the fourth month, but without complete resolution of the lesions.

Animals↗

Molecular epidemiology of Fonsecaea pedrosoi using mitochondrial DNA analysis.

Restriction fragment length polymorphism (RFLP) of mitochondrial DNA (mtDNA) from 49 clinical Fonsecaea pedrosoi isolates (18 isolates from Japan, 17 from Madagascar, 7 from Argentina, 5 from Venezuela, 1 from Costa Rica and 1 unknown) was studied. The 49 isolates were composed of 20 isolates of Type 1, 16 of Type 2, 12 of Type 4 and 1 of a new mtDNA type, Type 7, which was closely related to Type 2. On the bases of the results of 120 isolates of the present (49 isolates) and previous (71 isolates) studies, F. pedrosoi was classified into seven mtDNA types and according to the relationship between mtDNA types and geographic origins: in Japan and probably in China, Type 1 isolates; in Zaire and Madagascar, Type 2; in Central and South America, Type 4 and Type 1. These results indicated that the geographical origin of F. pedrosoi isolate could be roughly inferred from its mtDNA type.

Chromoblastomycosis↗

Mitochondrial DNA analysis of Sporothrix schenckii in North and South America.

Mitochondrial DNA (mtDNA) types based on restriction fragment length polymorphism (RFLP) patterns with HaeIII were investigated in clinical isolates of Sporothrix schenckii in North and South America. In addition to 14 mtDNA types (Types 1-14) so far reported, six new mtDNA types, Types 15-20 were found in this study. Type 3 was divided into two subtypes, Subtype 3A and Subtype 3B based on RFLP with Msp1. Type 14 was also divided into three subtypes, Subtype 14A, Subtype 14B and Subtype 14C based on RFLP with Hha1. Nineteen isolates in the United States consisted of 1 isolate of Type 1, 12 of Type 2, 2 of Type 4, 3 of Type 14 (1 of Subtype 14B and 2 of Subtype 14C) and 1 of Type 15. Twenty nine isolates in Venezuela consisted of 13 of Type 3 (Subtype 3B), 6 of Type 4, 1 of Type 18, 3 of Type 19 and 6 of Type 20. Thirteen isolates in Argentina consisted of 2 of Type 3 (Subtype 3A), 4 of Type 4, 4 of Type 16 and 3 of Type 17. One isolate in Brazil was Type 3 (Subtype 3A). Based on the phylogeny of 20 mtDNA types (Types 1-20) constructed by estimating sequence divergences of mtDNA, mtDNA types were clustered into two groups: Group A (Types 1-3, Type 11 and Types 14-19) and Group B (Types 4-10, Types 12-13 and Type 20). These results suggest that S. schenckii isolates in North and South America mainly belong to Group A.

DNA, Fungal↗

Acid fast filaments in stool samples from an AIDS patient.

The presence of filamentous bacteria morphologically similar to Nocardia in a fresh stool sample from an AIDS patient with pulmonary nocardiosis is here reported. The material was submitted to our laboratory for a parasitologic examination and was stained by the Kinyoun method, revealing numerous delicate, irregularly stained, branching acid-fast filaments. Nocardia asteroides had been isolated from sputum samples of this patient. The patient was a 32 year-old HIV+ female admitted to our center on June 1997 because of productive cough, right-sided thoracic pain and weight loss. Chest X rays showed the presence of right superior lobe excavated pneumonia. This was the first time we had observed filamentous bacteria similar to Nocardia in a stool sample submitted to parasitologic examination. For similar cases, and when its presence was not detected in other specimens collected from the same patient, intestinal endoscopy and biopsy should be performed for eventual lesions and smear examination repeated with Kinyoun stain and cultures for Nocardia.

AIDS-Related Opportunistic Infections↗

Teaching medical mycology in the year 2000.

Medical mycology is of increasing interest to the basic scientist, pathologist, microbiologist and clinician. This interest has been prompted by the rising number of immunosuppressed patients with opportunistic fungal infections, the expanding boundaries of the so-called endemic mycoses, the recognition of several major new endemic mycoses and a variety of other emerging fungal infections, and the development of potent, non-toxic antifungal drugs to treat these infections. The world of mycology is changing dramatically, especially in developing countries which have only limited resources to cope with the impact of the compromised host and the introduction of costly new antifungal drugs. Consequently, there is an urgent need to increase our effectiveness as teachers of medical mycology at all levels and in all regions of the world.

Education, Graduate↗

The role of ISHAM: how developing and developed countries can benefit from it. International Society for Human and Animal Mycology.

The international community of mycologists, ISHAM, is faced increasingly with the needs and demands of supporting the development of organizational structures worldwide in the field of medical mycology. This goes beyond the classical objectives of a scientific association, namely organizing international congresses and publishing a journal. In the early part of the next century, this will become a major challenge to ISHAM. Symposia discussing this subject were presented at the 12th Congress of ISHAM held in Adelaide, Australia, in 1994. This paper outlines the reflections and discussions held by members of our Society in 1997.

Animals↗