Paracocci

J Med Vet Mycol, 1995 Nov-Dec, 33(6), 411 - 4
Paracoccidioidomycosis in a renal transplant recipient; Shikanai-Yasuda MA et al.; Paracoccidioidomycosis is a fungal infection rarely described in immunodeficient patients . We report a severe case of pulmonary paracoccidioidomycosis in a renal transplant recipient and demonstrate deficiencies of in vitro lymphocytic transformation assays, skin hypersensitivity tests, as well as low levels of antibodies to Paracoccidioides brasiliensis.

J Med Vet Mycol, 1995 Nov-Dec, 33(6), 379 - 83
Immunochemical study of a Paracoccidioides brasiliensis polysaccharide-like antigen; Mendes-Giannini MJ et al.; The polysaccharide antigen from P . brasiliensis has been largely employed in serologic tests ,as well as in skin tests, to evaluate cellular immunity . SDS-PAGE analysis of this antigen has revealed a variability in the number of bands exhibited by isolates SN, 265, 339, 113, and 18 (7 to 16 bands) . The antigens obtained from isolates 2, PTL, 192 and Adel showed two or three bands . Glycoprotein analysis demonstrated a broad region between 50 and 90 kDa . Major bands of 48 and 30 kDa were present in almost all antigens . Optimal complement fixing dilution appears to be unaffected by the number of bands presented by different antigens . The immunoblot analysis revealed that the 90 and 30 kDa bands were mainly recognized by sera from paracoccidioidomycosis patients . Bands of high molecular weight were also recognized by most of the sera studied . Sera from histoplasmosis recognized the 94 kDa band . In conclusion, although the isolates exhibit quantitative variability in the number of fractions, it is possible to use only one or two samples given the greatest frequency of reactivity is seen in the 30 and 90 kDa fractions.

Clin Infect Dis, 1995 Nov, 21(5), 1275 - 81
Paracoccidioidomycosis and AIDS: an overview; Goldani LZ et al.; The scarcity of reported cases of paracoccidioidomycosis and AIDS remains unexplained . We review the details of the 27 cases reported in the medical literature . Paracoccidioidomycosis occurs in patients with advanced AIDS who are not receiving prophylaxis for Pneumocystis carinii pneumonia with trimethoprim-sulfamethoxazole, which is also effective against Paracoccidioides brasiliensis . Clinical manifestations include prolonged fever, weight loss, generalized lymphadenopathy, splenomegaly, hepatomegaly, and skin rash . Diagnosis can often be made by direct microscopic examination and culture of the fungus from skin and lymph node specimens and occasionally from sputum, blood, spinal fluid, and bone marrow specimens . Since antibodies to P . brasiliensis are occasionally detected, the diagnosis should not be ruled out for patients whose serology is negative . Despite specific therapy with different regimens, the overall mortality of paracoccidioidomycosis among patients with AIDS is high (30%) . The prognosis can be improved by earlier diagnosis and aggressive therapy with amphotericin B, followed by lifelong immunosuppressive therapy with trimethoprim-sulfamethoxazole . Health care providers caring for human immunodeficiency virus-infected patients who live or have resided in areas in which paracoccidioidomycosis is endemic must be aware of the possibility that this systemic mycosis may occur and have potentially severe consequences.

Rev Inst Med Trop Sao Paulo, 1995 Sep-Oct, 37(5), 407 - 13
Clinical and endoscopic findings in the mucosae of the upper respiratory and digestive tracts in post-treatment follow-up of paracoccidioidomycosis patients; do Valle AC et al.; Systematic examination of the upper respiratory and digestive tracts (URDT) was performed in a group of 80 paracoccidioidomycosis (PCM) patients submitted to post-treatment follow-up ranging from 8 months to 17 years . Mucosae of the URDT had been involved prior to specific treatment in 74 patients, distributed as follows: oropharynx, 50 (41 alone, 7 in association with the larynx, and 2 with the nasal mucosa); larynx, 30 (23 alone and 7 in association); and nasal mucosa, 3 (1 alone and 2 in association) . Inactive lesions were observed in all the 50 patients with lesions of the oropharynx, 3 of whom with deforming scars (1 with retraction of the tongue and 2 with narrowing of the oral orifice) . One case presented a destructive lesion, with perfuration of the palate . Of the other 46 cases, examination showed nacreous white striated scars which were nearly imperceptible in some cases and in others displayed partial retraction of anatomical structures without any alteration of their features . Patients presented a high rate of missing teeth . In 3 patients with involvement of the nasal mucosa, none of whom presented active PCM lesions, 2 still had nasal voices . In 30 patients with lesions of the larynx, 1 suffered a relapse of PCM and 2 developed epidermoid carcinoma . Of the other 27 cases, none of whom had active PCM lesions, 15 presented dysphonia, 3 were tracheotomized, and 9 were asymptomatic.

Rev Med Panama, 1995 Sep, 20(3), 65 - 71
{Dermatology in the tropics}; Tapia Collantes A; The Author reviews the cases diagnosed in the Republic of Panama of Mycetoma, Paracoccidioidosis, Lobo's disease, Chromomycosis, Histoplasmosis, Rhinosporidiosis, Sporotrichosis, Lepra, Rhinoscleroma, and cutaneous and mucocutaneous Leishmaniasis, and mentions the observed clinical manifestations in order to familiarize young physicians with the tropical dermatopathology which occurs in the rural areas of the country.

Trans R Soc Trop Med Hyg, 1995 Sep-Oct, 89(5), 566 - 72
Isolation and partial characterization of a Paracoccidioides brasiliensis 58 kDa extracellular glycoprotein which is recognized by human immune sera; Figueroa JI et al.; A novel 58 kDa antigenic determinant of the fungus Paracoccidioides brasiliensis was identified by enzyme-linked immunosorbent assay using a panel of species-specific murine monoclonal antibodies (MAbs) . Western immunoblot analysis, deglycosylation studies and isoelectric focusing indicated that this 58 kDa antigen is a glycoprotein, with a pI of approximately 5.2 . The molecule was purified from P . brasiliensis culture filtrate and yeast cytoplasmic antigens by membrane ultrafiltration, liquid isoelectric focusing and gel filtration; N-terminal amino acid sequence data revealed no substantial homology with known proteins . The presence of the antigen in the cytoplasm of both yeast and mycelial forms of the fungus was demonstrated when these MAbs were used as markers in immunofluorescence, immunoperoxidase and immunoalkaline phosphatase techniques to label P . brasiliensis in cryostat sections . These MAbs also recognized the cytoplasm of P . brasiliensis yeast forms in paraffin-embedded pathological specimens from human cases . A preparation of the 58 kDa component from yeast cytoplasmic antigen was reacted by Western immunoblotting with 26 different serum samples from paracoccidioidomycosis patients, and 81% of them recognized it.

J Med Vet Mycol, 1995 Sep-Oct, 33(5), 281 - 5
Isoenzyme profile of Paracoccidioides brasiliensis; Svidzinsky TI et al.; Isoenzyme profiles of 10 strains of Paracoccidioides brasiliensis from different origins (nine strains from patients with different clinical forms of paracoccidioidomycosis and one from the faeces of a penguin) were determined by polyacrylamide gel electrophoresis using 37 different enzymes . Differences in carbonate dehydratase, phosphoglucomutase, phosphoglucoseisomerase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, lactate dehydrogenase and beta-esterase were detected among the isolates studied allowing the characterization of nine zymodemes . Two isolates showed identical profiles . There was no correlation between the zymodeme patterns and virulence, clinical forms of the disease nor age of the cultures.

Am J Trop Med Hyg, 1995 Aug, 53(2), 189 - 94
Responses of T and B lymphocytes to a Paracoccidioides brasiliensis cell wall extract in healthy sensitized and nonsensitized subjects; Benard G et al.; Antigen-specific cellular immunity in paracoccidioidomycosis (PCM) has been poorly studied due to lack of standard in vitro lymphocyte proliferation assays . To standardize such an assay, we studied T and B cell responses to a Paracoccidioides brasiliensis cell wall extract (PbAg) in healthy subjects sensitized to either P . brasiliensis {Pb(+)Hc(-)} or to Histoplasma capsulatum {Hc(+)Pb(-)}, and in nonsensitized persons . All subjects showed, as expected, a vigorous proliferative response to a control fungal antigen obtained from Candida albicans . Lymphocytes from Pb(+)Hc(-) donors, but not from Pb(-)Hc(-) donors, reacted to PbAg by proliferating in a dose-dependent manner with a maximum reaction after 6-9 days, suggesting a secondary specific immune response . Most activated cells were CD+CD4+ lymphocytes . However, Hc(+)Pb(-) donors' cells reacted with PbAg . Cross-reactivity with H . capsulatum was not unexpected, since both fungi, but not C . albicans, share cell wall immunogenic compounds . An enzyme-linked immunosorbent assay to detect human immunoglobulins (Ig) demonstrated that B cells from Pb(+)Hc(-) donors, but not from Pb(-)Hc(-) ones, reacted with PbAg by secreting high levels of IgG and IgM in 12-day culture supernatants . This secretion was possibly mediated by PbAg-activated CD4+ cells . We believe that analysis of T and B lymphocyte responses to PbAg will be useful in the investigation of the infection-associated immune impairment seen in some PCM patients.

Clin Exp Immunol, 1995 Aug, 101(2), 321 - 7
The role of somatic structure of the fungus Paracoccidioides brasiliensis upon B cell activation in experimental paracoccidioidomycosis; Silva MF et al.; In this study, we report an increase of the number of antibody-secreting cells and the augmentation of antibody production against unrelated antigens in mice infected with the fungus P . brasiliensis, as well as in mice inoculated with cell wall preparation isolated from P . brasiliensis (CW) . The immunomodulatory effect of the live fungus and the CW preparation was dose-dependent, and their actions were mainly restricted to the i.v . or i.p . inoculation simultaneously with the sheep erythrocyte challenge by the i.v . route or restricted to i.p . inoculation of CW when bovine serum albumin (BSA) antigen was used . The dependence of antibody production on different routes of CW inoculation was correlated with the number of antigen-specific B cells in the spleen as determined by direct and reverse plaque-forming cell assays . The immunization schedules using CW preparation caused a preferential production of IgM and IgG3 antibodies . The results also showed that the hyperactive humoral immune response of mice induced by i.p . inoculation of CW was devoid of polyclonal B cell activation compared with the effects observed for the lipopolysaccharide (LPS)-treated groups . Paracoccidioides brasiliensis CW components may have potent immunological properties related to the non-specific B cell activation found in paracoccidioidomycosis.

Clin Exp Immunol, 1995 Aug, 101(2), 314 - 20
Differential correlation between interleukin patterns in disseminated and chronic human paracoccidioidomycosis; Silva CL et al.; In an attempt to understand better the immunoregulatory disorders in paracoccidioidomycosis (PCM), the possible correlation between interleukin pattern, lymphoproliferation, C-reactive protein (CRP) and specific antibody levels was investigated in the polarized clinical forms of this disease . We studied 16 PCM patients, eight with the disseminated disease (four under treatment and four non-treated) and eight with the chronic disease . The patients with disseminated disease exhibited high antibody titres specific to Paracoccidioides brasiliensis antigen compared with patients with the chronic form of disease . Tumour necrosis factor (TNF), IL-1, IL-6 and CRP in the serum of non-treated disseminated PCM patients were increased, which correlated positively with the low mitogenic response of peripheral blood mononuclear cells (PBMC) to phytohaemagglutinin (PHA) (P < 0.01) and with the high antibody titres (P < 0.001) of these patients . Moreover, we found in the disseminated PCM patients positive correlations between IL-1 and IL-6 (P = 0.0007); IL-1 and TNF (P = 0.0045); IL-1 and IL-6 with the high antibody titres (P = 0.0834 and P = 0.0631, respectively); IL-1, IL-6 and TNF with CRP levels . By contrast, no correlations were found with those interleukins in the treated disseminated and chronic patients or in controls . It was interesting to find an inverse correlation between IL-4 and antibody production in non-treated disseminated PCM (r = -0.4770); moreover, a significant correlation (P = 0.0820) was found in chronic PCM patients with respect to the low level of either IL-4 and antibody titres against fungus antigen . Chronic PCM patients also had IL-2 levels inversely correlated with antibody production (r = -0.6313; P = 0.0628) . Inverse correlations were also observed between IL-2 and IL-6 levels in non-treated disseminated patients (P = 0.0501) and between IL-2 and IL-4 in chronic patients (P = 0.0131) . The inflammatory cytokines might have a pivotal role in the genesis and in control of some aspects of the disease, such as granulomatous reaction, hypergammaglobulinaemia and depression of T cell-mediated immunity in PCM.

Rev Inst Med Trop Sao Paulo, 1995 Jul-Aug, 37(4), 303 - 9
The hamster cheek pouch: an immunologically privileged site suitable to the study of granulomatous infections; de Arruda MS et al.; The hamster cheek pouch is an invagination of oral mucosa, characterized histologically as skin-like . In this paper we describe anatomical, histological and embriological features of the pouch and comment on the pouch as an immunologically privileged site since it lacks lymphatic drainage and has few Langerhans cells . We present the review from literature and our observations after inoculation in the pouch of mycobacteriae (BCG, Mycobacterium tuberculosis and Mycobacterium leprae) and a fungus (Paracoccidioides brasiliensis) . Lesions in the pouch were granulomatous but smaller and long lasting; even granulomatous, the reaction was inefficient to control the proliferation of agents compared with inoculation in other sites, except for BCG . Appearance of immunity was also delayed or absent and, when it was detected, a sharp decrease in number of agents in pouch lesions was observed . These observations make the pouch an interesting site for the study of the role of immune system in infectious diseases and in granuloma formation.

Rev Argent Microbiol, 1995 Jul-Sep, 27(3), 139 - 45
{Isolation of an Paracoccidioides brasiliensis exoantigen from solid culture media}; Gago J et al.; The goal of this work was to develop in solid medium a fast method to obtain Paracoccidioides brasiliensis (Pb) with a high yield . Four culture media were assayed: Sabouraud honey-agar, Sabouraud dextrose-agar, tomato -agar-medium (TOM) and a medium based on grape pulp . The most exhuberant growth was observed in medium based on grape pulp . Antigen was prepared in microscale at 6, 10 and 15 days incubation of solid cultures and the crude product concentrated by means of Centriplus tubes (Helena, France) . Isolated antigens were subjected to polyacrylamide gel electrophoresis, followed by immunolabelling and detection of the characteristic gp45 antigen employing human and Pb-infected rat sera . Best results were observed after 10 days culture in grape medium . None of the other three media afforded comparable results.

J Med Vet Mycol, 1995 Jul-Aug, 33(4), 247 - 51
Glycolipids from Paracoccidioides brasiliensis . Isolation of a galactofuranose-containing glycolipid reactive with sera of patients with paracoccidioidomycosis; Toledo MS et al.; In the present study, we describe the isolation of glycolipids from yeast and mycelium forms of Paracoccidioides brasiliensis . Both forms contains glucosylceramide as the only neutral glycosphingolipid and two acidic glycolipids termed band 1 and band 2 . Band 1 was found to be reactive with 100% of sera of patients with paracoccidioidomycosis tested . Structural analysis of band 1 revealed that it is composed of mannose and galactose in molar ratios of 2:1, and a trace amount of glucose . Furthermore, this paper presents evidence that the galactose unit of band 1 is in the furanose configuration . Finally, it was found that reactivity of paracoccidioidomycosis sera with band 1 glycolipid can be attributed mainly to antibodies directed to galactofuranosyl residue present in this glycoconjugate.

J Med Vet Mycol, 1995 Jul-Aug, 33(4), 223 - 7
Expression and isoforms of gp43 in different strains of Paracoccidioides brasiliensis; Campos MC et al.; Individual exoantigens from 13 Paracoccidioides brasiliensis isolates were obtained and tested against a panel of 50 sera from patients with paracoccidioidomycosis by immunodiffusion test . The index of positivity varied from 20 to 100% according to the isolate . When these exoantigens were analysed by SDS-PAGE, eight presented high amounts of the glycoprotein gp43, two presented small amounts and in three there was no detectable gp43 . The eight isolates presenting high amounts of gp43 were submitted to isoelectric focusing, blotted to nitrocellulose membranes, and revealed by monoclonal and polyclonal anti-gp43 antibodies . Four gp43 isoform profiles were obtained: profile A presenting pIs of 6.0, 6.2, 6.6 and 7.0, profile B presenting pIs of 6.4, 6.8 and 7.2, profile C presenting pI > 8.5 and profile D, presenting pIs of 5.8, 6.2 and 6.6 . In each pattern observed, a major band with a distinct pI was detected . Despite this variation, our results strongly suggest an epitopic conservation among all isoforms analysed, since all of them were recognized by anti-gp43 monoclonal antibodies.

Clin Exp Immunol, 1995 Jul, 101(1), 114 - 20
Influence of the genetic pattern and sex of mice in experimental paracoccidioidomycosis; Singer-Vermes LM et al.; Eight genetically different strains of mice were compared regarding the dissemination of Paracoccidioides brasiliensis to the lungs, liver and omentum/pancreas, DTH responses and specific antibody production at 16 weeks after intraperitoneal infection with Pb18, a virulent P . brasiliensis isolate . The degree of dissemination of the infection varied: B10.A and C57B1/6, the most susceptible mouse strains, had positive cultures and high colony-forming unit (CFU) counts in all analysed organs . DBA/2 and A/Sn mice had negative cultures, being thus classified as the most resistant strains . CBA/J, C3H/HeJ, F1(A/SnxB10.A) and BALB/c mice were regarded as relatively resistant, since discrete fungal growth was observed only in one or two of the studied organs . All mouse strains, except B10.A mice, produced specific DTH responses which did not seem to be associated with the severity of disease . Production of high levels of specific antibodies was found in all strains except in the DBA/2 and C57B1/6 mice . The influence of the host sex on the outcome of paracoccidioidomycosis was evident only in susceptible animals: female B10.A mice displayed lower CFU counts in the three examined organs, whereas no differences were found between male and female A/Sn animals . The higher resistance of female B10.A mice was not accompanied by differences in their capacity to maintain a DTH reaction, nor in their production of antibody . This fact argues against the widely believed association of susceptibility to P . brasiliensis infection with both impaired DTH reactivity and increased humoral response.

Biochem J, 1995 Jul 1, 309 ( Pt 1), 209 - 14
Characterization of an exocellular serine-thiol proteinase activity in Paracoccidioides brasiliensis; Carmona AK et al.; An exocellular proteinase activity has been characterized in Paracoccidioides brasiliensis culture filtrates . Chromatographic analysis showed that the activity was eluted from an anion-exchange Resource Q column at 0.08-0.1 M NaCl, and by gel filtration near ovalbumin elution, in a single peak . Purification of the proteinase, however, was hampered by the low protein yield, in contrast to the high peptidase activity . Numerous chromogenic peptidyl p-nitroanilide derivatives and internally quenched fluorescent peptides, flanked by Abz (O-aminobenzoyl) and EDDnp (ethylenediaminedinitrophenyl), were tested as substrates . Cleavage was observed with Abz-MKRLTL-EDDnp, Abz-FRLVR-EDDnp, and Abz-PLGLLGR-EDDnp at Leu-Thr, Leu-Val and Leu-Leu/Leu-Gly bonds respectively as determined by isolation of the corresponding fragments by HPLC . Leucine at P1 seemed to be restrictive for the activity of the exocellular enzyme, but threonine (P'1) and leucine (P'2) in Abz-MKRLTL-EDDnp apparently were not essential . Also, a pair of alanines could substitute for lysine (P3) and arginine (P2) in this substrate, with a decrease in the Km values . The exocellular peptidase activity of P . brasiliensis had an optimum pH of > 9.0 and was irreversibly inhibited by PMSF, mercuric acetate and p-hydroxymercuribenzoate . Inhibition of the mercuriate compounds could be partially reversed by Cys/EDTA . E-64 {trans-epoxysuccinyl-L-leucylamido-(4-guanido)butene} was a weak and reversible inhibitor, whereas EDTA and pepstatin were not inhibitory . These results suggest that P . brasiliensis exocellular enzyme belongs to the subfamily of SH-containing serine proteinases.

Diagn Cytopathol, 1995 Jul, 13(1), 52 - 3
Paracoccidioidomycosis: diagnosis by fine-needle aspiration cytology; Drut R; A 4-yr-old girl presented with constitutional symptoms, abdominal swelling, ascites, and cervical lymphadenopathy . Fine-needle aspirate smears of a cervical lymph node revealed numerous round yeasts, many of them with several peripheral buds fitting the pattern of Paracoccidiodes brasilensis . This appears to be the first case in which the diagnosis of this deep mycosis was achieved by FNAC . The procedure may prove useful for rapid diagnosis in cases like the present one which represents the so-called acute disseminated form which affects mainly children and immunosuppressed patients and can be rapidly fatal.

Rev Soc Bras Med Trop, 1995 Jul-Sep, 28(3), 279 - 84
{Paracoccidioidomycosis in a patient with human immunodeficiency virus infection . A necropsy case}; de Lima MA et al.; This is a case report of the association of Paracoccidioidomycosis and Acquired Immunodeficiency Syndrome (AIDS) occurring in a 43-year old male . This is, to the best of our knowledge, the first detailed pathological account of that association . Also discussed are the low rates of that association, its natural history and treatment results . It is emphasised the importance of the associations of AIDS and tropical infectious diseases in this country.

Exp Mycol, 1995 Jun, 19(2), 111 - 9
Paracoccidioides brasiliensis expresses both glycosylphosphatidylinositol-anchored proteins and a potent phospholipase C; Heise N et al.; This study reports, for the first time, the detection of glycosylphosphatidylinositol (GPI) membrane anchors in proteins of a pathogenic fungus, Paracoccidioides brasiliensis . Taking into account that fungal antigens are found in the sera of paracoccidioidiomycosis patients and that cleavage of this glycolipid by phospholipases is a means of selective protein release, the presence of an enzyme with this property has also been investigated . Using a methodological approach in which the proteins were immobilized on nitrocellulose, treated with phospholipase C of Trypanosoma brucei and then probed with antibodies which recognize the 1,2-cyclic-phosphate inositol moiety formed as a reaction product in proteins bearing the glycolipid anchor, it was possible to detect a major glycoprotein in the 80- to 90-kDa range, as well as two other minor species of 66 and 43 kDa . All of them bind to Concanavalin-A and are also substrates of a very potent fungal phospholipase C which is inhibited by p-chloromercuri-phenylsulfonic acid and is insensitive to EDTA . The integrity of glycosylphosphatidylinositol anchors in proteins of P . brasiliensis is impaired by 0.1 M NaOH, a finding indicative of a diacyl glycerolipid moiety which is quite surprising since it is, with the exception of African trypanosomes surface proteins and Torpedo acetylcholinesterase, an uncommon feature among GPIs in general . The present findings may have implications in the pathology of paracoccidiodomycosis.

Mycopathologia, 1995 Jun, 130(3), 131 - 40
Effect of macrophage blockade on the resistance of inbred mice to Paracoccidioides brasiliensis infection; Kashino SS et al.; The effect of macrophage blockade on the natural resistance and on the adaptative immune response of susceptible (B10.D2/oSn) and resistant (A/Sn) mice to Paracoccidioides brasiliensis infection was investigated . B10.D2/oSn and A/Sn mice previously injected with colloidal carbon were infected ip with yeast cells to determine the 50% lethal dose, and to evaluate the anatomy and histopathology, macrophage activation, antibody production and DTH reactions . Macrophage blockade rendered both resistant and susceptible mice considerably more susceptible to infection, as evidenced by increased mortality and many disseminated lesions . P . brasiliensis infection and/or carbon treatment increased the ability of macrophages from resistant mice to spread up to 25 days after treatment . In susceptible mice the enhanced spreading capacity induced by carbon treatment was impaired at all assayed periods except at 1 week after infection . Macrophage blockade enhanced DTH reactions in resistant mice, but did not alter these reactions in susceptible mice, which remained anergic . To the contrary, macrophage blockade enhanced specific antibody production by susceptible mice, but did not affect the low levels produced by resistant mice . The effect of macrophage blockade confirms the natural tendency of resistant animals to mount DTH reactions in the course of the disease and the preferential antibody response developed by susceptible mice after P . brasiliensis infection . On the whole, macrophage functions appear to play a fundamental role in the natural and acquired resistance mechanisms to P . brasiliensis infection.

Rev Inst Med Trop Sao Paulo, 1995 May-Jun, 37(3), 261 - 5
Paracoccidioidomycosis associated with acquired immunodeficiency syndrome . Report of seven cases; Marques SA et al.; We report the clinical findings and evolution of seven patients (five men and two women), the majority of them intravenous drug users, with paracoccidioidomycosis associated to acquired immunodeficiency syndrome (AIDS) . In four of the patients the paracoccidioidomycosis was restricted to the lung and in the three others was generalized with cutaneous involvement . Only two of them had lived recently in rural area, an indication of the possible reactivation of latent focal infection in the other five patients . The recognition of the role of cell-mediated immunity in host defense against Paracoccidioides brasiliensis leds to the prediction of a growing occurrence of the paracoccidioidomycosis-AIDS association in areas that are endemic for these diseases.

Rev Inst Med Trop Sao Paulo, 1995 May-Jun, 37(3), 219 - 24
Cyclophosphamide effect on paracoccidioidomycosis in the rat; Blejer JL et al.; Paracoccidioidomycosis is an endemic fungal disease widely distributed throughout Latin America . The potent immunosuppressor cyclophosphamide (CY) has been used to modulate host immune response to Paracoccidioides brasiliensis in an experimental model . Inbred male Buffalo/Sim rats weighing 250-300 g were inoculated with 5 x 10(6) P . brasiliensis cells of the yeast phase form by intracardiac route . One group of animals was treated with 20 mg/kg body weight at days +4, +5, +6, +7, +11 and +12 post-infection (pi.), while a control group was infected alone . No mortality was recorded in either group . Treated rats presented: a) a decrease in granuloma size, which contained less fungal cells; b) a lack of specific antibodies up to 35 days pi., and c) a significant increase in the footpad swelling test (DTH) against paracoccidioidin . Splenic cell transfer from CY-treated P . brasiliensis-infected donors to recipients infected alone led to a significant increase in DTH response in the latter versus untreated infected controls . Likewise, in treated infected recipients transferred with untreated infected donor spleen cells, footpad swelling proved greater than in controls . Thus, it would seem that each successive suppressor T lymphocyte subset belonging to the respective cascade may be sensitive to repeated CY doses administered up to 12 days pi. . Alternatively, such CY schedule may induce the appearance of a T cell population capable of amplifying DTH response.

Infect Immun, 1995 May, 63(5), 1777 - 83
Pulmonary paracoccidioidomycosis in resistant and susceptible mice: relationship among progression of infection, bronchoalveolar cell activation, cellular immune response, and specific isotype patterns; Cano LE et al.; Using the intraperitoneal route of infection, we demonstrated previously that A/Sn mice are resistant and B10.A mice are susceptible to Paracoccidioides brasiliensis infection . Since paracoccidioidomycosis is a deep systemic granulomatous disorder that involves primarily the lungs and then disseminates to other organs and systems, we herein investigated the course of the infection and the resulting immune responses developed by A/Sn and B10.A mice after intratracheal infection with P . brasiliensis yeast cells . It was observed that A/Sn mice develop a chronic benign pulmonary-restricted infection, whereas B10.A mice present a chronic progressive disseminated disease . A/Sn animals were able to restrict fungal infection to the lungs despite the increased fungal load at the beginning of the infection . This behavior was associated with low mortality rates, the presence of adequate and persistent delayed-type hypersensitivity reactions, oxidative burst by bronchoalveolar cells, and production of high levels of specific antibodies in which immunoglobulin G2a (IgG2a) and IgG3 isotype titers were significantly higher than those observed in the susceptible mice . In contrast, B10.A animals showed a constant pulmonary fungal load and dissemination to the liver and spleen . This infection pattern resulted in high mortality rates, discrete delayed-type hypersensitivity reactivity, poorly activated or nonactivated bronchoalveolar cells, and production of specific IgG2b isotype titers significantly higher than those observed in the resistant mice at week 4 of infection . Thus, A/Sn and B10.A mice maintain the same resistance patterns as those observed previously with the intraperitoneal route of infection . Furthermore, the obtained results suggest that resistance to paracoccidioidomycosis is associated with T-cell, macrophage, and B-cell activities that are known to be mediated by gamma interferon.

Mycopathologia, 1995 May, 130(2), 75 - 8
Polyserositis in a patient with acute paracoccidioidomycosis and hepatosplenic schistosomiasis; Shikanai-Yasuda MA et al.; A severe case of juvenile paracoccidioidomycosis (PCM), manifested as cholestatic jaundice, lymph node enlargement and an unusual form of polyserositis, associated with portal hypertension secondary to schistosomiasis, as well as bacteremias caused by E . coli and S . aureus and post-transfusional hepatitis C is reported . Temporary unresponsiveness of in vivo and in vitro cellular immune responses to P . brasiliensis were registered . The authors discuss the possible interference of either agent in the host immune response, thus explaining the severity of PCM in the present case.

Infect Immun, 1995 Apr, 63(4), 1608 - 10
Paracoccidioides brasiliensis-stimulated human gamma/delta T cells support antibody production by B cells; Munk ME et al.; Paracoccidioidomycosis patients show hyperactive humoral immune responses . Consequently, we investigated whether cytokines in supernatants from Paracoccidioides brasiliensis-stimulated gamma/delta T cells support B-cell activation . We detected proliferation of B cells and increased immunoglobulin M (IgM) and IgG production . Thus, gamma/delta T cells may participate in polyclonal B-cell activation during paracoccidioidomycosis.

J Med Vet Mycol, 1995 Mar-Apr, 33(2), 113 - 6
Lack of reactivity of paracoccidioidomycosis sera in the double immunodiffusion test with the gp43 antigen: report of two cases; del Negro GM et al.; Sera from two patients with chronic active paracoccidioidomycosis yielded negative double immunodiffusion results with a culture filtrate antigen from Paracoccidioides brasiliensis routinely used in our laboratory . Complement fixation tests were positive for both sera using a polysaccharide-rich antigen . This study reports the results of a more extensive serological investigation of these two sera . Both a somatic antigen and a saline extract from the fungus yielded positive results in the double immunodiffusion . However, the immunodominant 43 kDa glycoprotein antigen showed negative results, although it was recognized by both sera in the Western blot assay . The value of the double immunodiffusion as a single serological test in paracoccidioidomycosis diagnosis is discussed.

Rev Inst Med Trop Sao Paulo, 1995 Mar-Apr, 37(2), 129 - 36
Natural killer cell activity in experimental paracoccidioidomycosis of the Syrian hamster; Peracoli MT et al.; The study evaluated the activity of NK cells during the course of experimental infection of hamsters with Paracoccidioides brasiliensis . Eighty hamsters were infected with P . brasiliensis by intratesticular route and sacrificed at 24h, 48h, 96h, 1, 2, 4, 8 and 11 weeks of infection and compared to 40 noninfected hamsters employed as controls . These animals were submitted to the study of NK cytotoxic activity by a single-cell assay and humoral immune response by immunodiffusion and ELISA tests . The production of macrophage migration inhibitory factor in the presence of Phytohemagglutinin and P . brasiliensis antigen and histopathology of the lesions were evaluated at 1, 4, 8 and 11 weeks of infection . The infected animals displayed significantly high levels of NK activity during the four weeks of infection that decreased from the 8th week on when compared to controls . This impairment of NK activity was associated with depression of cell-mediated immune response and with increase in the extension of the histopathologic lesions . There was an inverse correlation between NK cell activity and specific antibody levels . The results suggest that after initial activation, NK cells were unable to control the fungus dissemination . The impairment of NK activity in the late stages of the infection might be related to immunoregulatory disturbances associated with paracoccidioidomycosis.

J Clin Microbiol, 1995 Feb, 33(2), 505 - 7
Characterization of Paracoccidioides brasiliensis isolates by random amplified polymorphic DNA analysis; Soares CM et al.; We initially used 25 different random primers in order to test their ability to generate random amplified polymorphic DNA fragments from the dimorphic human pathogenic fungus Paracoccidioides brasiliensis . From the tested primers we chose five to distinguish between seven isolates of this microorganism . The DNA amplification patterns allowed clear differentiation of the seven isolates into two distinct groups with only 35% genomic identity . One of these groups contained two subgroups with 81% genetic similarity . The random amplified polymorphic DNA analysis method proved to be a good tool for analyzing and comparing different genomes of P . brasiliensis isolates.

Braz J Med Biol Res, 1995 Feb, 28(2), 209 - 12
Granulomatous reaction in the hamster cheek pouch induced by killed Paracoccidioides brasiliensis; Arruda MS et al.; We have studied the role of Paracoccidioides brasiliensis viability in the morphology of paracoccidioidomycotic granulomas in the hamster cheek pouch, an immunologically privileged site . Naive (N = 75) and previously sensitized (N = 50) two-month old male hamsters (Mesocricetus auratus) were inoculated into the pouch with 5 x 10(5) live or heat- or formalin-killed fungi . Previously sensitized animals presented a positive footpad test and immunodiffusion demonstrable antibodies (titer 1/32), at the time of sacrifice; naive animals were always negative for those immunological tests . The histological results showed that, like viable P . brasiliensis, killed fungi evoke typical epithelioid granulomas in 100% of animals, even in the absence of immunodiffusion or footpad test detectable immune response . The granulomas elicited by killed fungi were devoid of giant cells or a mononuclear cell halo, suggesting that live proliferating fungi or their products may be involved in these events.

Arch Med Res, 1995 Autumn, 26(3), 305 - 6
In search of the natural habitat of Paracoccidioides brasiliensis; McEwen JG et al.; Paracoccidioides brasiliensis (Pb) is the dimorphic fungus responsible for paracoccidioidomycosis (PCM), one of the most important systemic mycosis in Latin America where the disease is geographically restricted . The natural habitat of the causative agent remains undetermined . We are planning to use PCR-based technology in order to amplify specific DNA fragments . The high sensitivity of this technique may allow us to detect the natural habitat of Pb . In this study, we prepared a cDNA library from which we cloned a protein of approximately 27 kDa MW . When this recombinant antigenic protein was tested by the immunoblot technique, it was able to recognize antibodies in the sera of 91% of the PCM patients studied . No cross reactions were observed with sera from patients with other systemic mycoses . Presently we are sequencing and characterizing this clone, in order to design specific primers for amplification of Pb DNA.

Arch Med Res, 1995 Autumn, 26(3), 297 - 304
Biochemistry and molecular biology of the main diagnostic antigen of Paracoccidioides brasiliensis; Travassos LR et al.; The 43,000 dalton glycoprotein of Paracoccidioides brasiliensis (gp 43) is the main exocellular antigen recognized by sera from patients with paracoccidioidomycosis in a variety of serological assays . Specific conformational peptide epitopes are recognized by the human antibodies as determined by antigen deglycosylation . Procedures for the purification of the gp43 using immunoaffinity chromatography have been described . The secretion of the gp43 as a function of the growth curve, its partial aggregation with a proteolytic enzyme, ability to bind laminin, as well as to form circulating immunocomplexes in vivo could play a role in pathogenesis . Crude antigenic preparations depleted of gp43 epitopes lost their ability to elicit positive skin tests . Accordingly, the purified gp43 molecule induced delayed hypersensitivity reactions in man and infected animals, caused a T-CD4-dependent proliferation of lymph node cells from mice immunized with it, and of peripheral blood lymphocytes from an individual sensitized to P . brasiliensis by prolonged contact with the fungus . To identify the immunodominant epitopes in both humoral and cellular reactions, the gp43 gene has been cloned, sequenced, and partly expressed . It bears peptide sequences homologous to those of beta-1,3-glucanases from Candida albicans and Saccharomyces cerevisiae but has no enzymatic activity itself . The molecular weight of the unglycosylated antigen is 42,227 . A single N-linked oligosaccharide chain in the gp43 contains alpha-D-mannopyranosyl, beta-D-galactofuranosyl and N-acetylglucosaminyl units with the predominant ratio of 10:2:2, and characteristics of a high mannose type.

Mem Inst Oswaldo Cruz, 1995 Jan-Feb, 90(1), 45 - 9
Optimization of a mouse immunization protocol with Paracoccidioides brasiliensis antigens; Almeida AV et al.; The objectives of the present study were to optimize the protocol of mouse immunization with Paracoccidioides brasiliensis antigens (Rifkind's protocol) and to test the modulation effect of cyclophosphamide (Cy) on the delayed hypersensitivity response (DHR) of immunized animals . Experiments were carried out using one to four immunizing doses of either crude particulate P . brasiliensis antigen or yeast-cell antigen, followed by DHR test four or seven days after the last immunizing dose . The data demonstrated that an immunizing dose already elicited response; higher DHR indices were obtained with two or three immunizing doses; there were no differences between DHR indices of animals challenged four or seven days after the last dose . Overall the inoculation of two or three doses of the yeast-cell antigen, which is easier to prepare, and DHR test at day 4 simplify the original Rifkind's immunization protocol and shorten the duration of the experiments . The modulation effect of Cy on DHR was assayed with administration of 2.5, 20 and 100 mg/kg weight at seven day intervals starting from day 4 prior to the first immunizing dose . Only the treatment with 2.5 mg Cy increased the DHR indices . Treatment with 100 mg Cy inhibited the DHR, whereas 20 mg Cy did not affect the DHR indices . Results suggest an immunostimulating effect of low dose of Cy on the DHR of mice immunized with P . brasiliensis antigens.

Immunology, 1995 Jan, 84(1), 98 - 104
Human cord blood T-cell receptor alpha beta cell responses to protein antigens of Paracoccidioides brasiliensis yeast forms; Munk ME et al.; Paracoccidioides brasiliensis causes a chronic granulomatous mycosis, prevalent in South America, and cell-mediated immunity represents the principal mode of protection against this fungal infection . We investigated the response of naive cord blood T cells to P . brasiliensis lysates . Our results show: (1) P . brasiliensis stimulates T-cell expansion, interleukin-2 (IL-2) production and differentiation into cytotoxic T cells; (2) T-cell stimulation depends on P . brasiliensis processing and major histocompatibility complex (MHC) class II expression; (3) the responsive T-cell population expresses alpha beta T-cell receptors (TCR) with different V beta gene products, CD4 and CD45RO; (4) the P . brasiliensis components involved in T-cell expansion primarily reside in a high molecular weight (100,000 MW) and a low molecular weight (< 1000 MW) protein fraction . These results indicate that protein antigens of P . brasiliensis stimulate cord blood CD4 alpha beta T cells, independent from in vivo presensitization, and thus question direct correlation of positive in vitro responses with protective immunity in vivo.

J Leukoc Biol, 1995 Jan, 57(1), 101 - 9
Comparative ultrastructure and immunolabeling of MHC-II antigens of alveolar macrophages obtained from patients with paracoccidioidomycosis and other lung diseases; Bretana A et al.; Samples of alveolar macrophages (AM) obtained by bronchoalveolar lavage from patients with either paracoccidioidomycosis, silicosis, sarcoidosis, or allergic alveolitis were investigated by electron microscopy and immunocytochemistry to compare cellular ultrastructure and expression of MHC-II antigens in the AM cell surface . All samples of AM obtained from patients with these pathologies showed heterogeneous structural features . Although, this morphological diversity is also present in AM of healthy donors, our observations seem to indicate that in the diseases studied this morphofunctional diversity is associated with additional ultrastructural characteristics inherent to each disease . In paracoccidioidomycosis the proportion of vacuolated macrophages is significantly lower than in other diseases; this might indicate that in paracoccidioidomycosis the proportion of activated AM is smaller . We observed significant differences in the expression of MHC-II antigens . Silicosis, sarcoidosis, and allergic alveolitis do not differ significantly in the quantity of immunolabeled AM or in the distribution of the label . The percentage of AM from paracoccidioidomycosis that exhibit the MHC-II molecule is very low with poor immunolabeling . In this disease the low expression of the MHC-II molecule could be related to a decrease of the antigen presenting function by AM.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 67 - 71
Severe juvenile type paracoccidioidomycosis in an adult; Benard G et al.; Paracoccidioidomycosis has recently been classified into juvenile (acute) and adult (chronic) forms . The latter affects middle-aged men and causes mucocutaneous lesions, while the more rare juvenile form affects the reticuloendothelial system of children and adults of both sexes under 30 years of age . It is not yet known, however, if the patient's age has a role in determining the immune response patterns to the fungus and the evolution to one form or the other . We present a 45-year-old man who presented with juvenile type disease characterized by intra-abdominal polyadenopathy forming a large epigastric mass . Immune evaluation showed high titres of anti-Paracoccidioides brasiliensis antibodies and an antigen-specific cellular immune defect . Treatment resulted in resolution of the clinical and immune abnormalities . His epidemiological background also suggested acute disease: he developed disease after moving from an urban to a rural endemic area . We suggest that acute or juvenile disease may occur in a previously healthy, susceptible individual when moving to an endemic area, at whatever age.

J Med Vet Mycol, 1995 Jan-Feb, 33(1), 39 - 42
Delayed hypersensitivity test with paracoccidioidin in captive Latin American wild mammals; Costa EO et al.; The aim of this investigation was to study epidemiological aspects of paracoccidioidomycosis, the main endemic systemic mycosis in Brazil . This study was carried out using the paracoccidioidin delayed hypersensitivity test in 96 Latin American wild mammals, including 49 arboreal animals (primates): 33 Cebus apella (weeping-capuchin), 16 Callithrix jacchus (marmoset); and 47 terrestrial animals (carnivora): 37 Nasua nasua (coatimundi), and 10 Felidae {Panthera onca (jaguar), Felis paradalis (ocelot), Felis wiedii (margay), Felis tigrina (wild cat) and Felis geoffroyi (wild cat)}, taking their behaviour and habitat into consideration . When the levels of paracoccidiodin positive reactions were examined, terrestrial animals showed significantly higher rates (82.98%) while arboreal animals showed lower reactivity (22.45%) (P < 0.01) . The data are relevant because there are quite a few papers regarding domestic and wild animals and this study may help the understanding of some aspects of the parasite ecology . These results point to the soil as the most probable reservoir of Paracoccidioides brasiliensis, and this is possibly the ecological niche of the saprophytic phase in nature.

Mycopathologia, 1995, 129(1), 17 - 23
Histoplasmosis in northwestern Argentina . II: Prevalence of Histoplasmosis capsulati and paracoccidioidomycosis in the population south of Chuscha, Gonzalo and Potrero in the province of Tucuman; van Gelderen de Komaid A et al.; The present work was undertaken to obtain epidemiological data on the extent and distribution of Histoplasma capsulatum var . capsulatum and Paracoccidiodes brasiliensis infections south of the Chuscha, Gonzalo and Potrero areas of Argentina . Skin tests surveys of the human population with histoplasmin and paracoccidioidin were carried out in the permanent population of those localities . The infection index of the population showed that the area south of Chuscha has a high-prevalence of histoplasmosis capsulati . The Gonzalo and Potrero areas, according to their rates of infection also can be considered to have a relatively high prevalence of this disease . The frequency of individuals infected with P . brasiliensis suggests that the level of exposure to this fungus is considerable, especially in Gonzalo where the frequency of infection was 9.23% . The endemic areas of both diseases can be superimposed, as occurs in the northeastern part of Argentina.

Rev Inst Med Trop Sao Paulo, 1995 Jan-Feb, 37(1), 43 - 9
Paracoccidioides brasiliensis . A mycologic and immunochemical study of a sample isolated from an armadillo (Dasipus novencinctus); Vidal MS et al.; A sample of P . brasiliensis isolated from the spleen and the liver of an armadillo (Dasipus novencinctus) has been analysed under a mycological and immunochemical viewpoint . The armadillo was captured in an area of Tucurui (State of Para, Brazil), the animal being already established as an enzootic reservoir of P . brasiliensis at that region of the country . This sample maintained in the fungal collection of the Tropical Medicine Institute of Sao Paulo (Brazil) numbered 135, has got all the characteristics of P . brasiliensis, with a strong antigenic power and low virulence for guinea-pigs and Wistar rats . The specific exoantigen of P . brasiliensis--the glycoprotein with a molecular weight of 43 kDa--was easily demonstrated with double immunodiffusion, immunoelectrophoresis, SDS-PAGE and immunobloting techniques.

Int Immunol, 1994 Nov, 6(11), 1717 - 25
Human alpha beta and gamma delta T cells from unexposed individuals respond to protein antigens of the yeast form of Paracoccidioides brasiliensis; Munk ME et al.; Paracoccidioides brasiliensis, a dimorphic fungus, causes chronic granulomatous mycosis in susceptible individuals . Different reports have shown that cell-mediated immunity is essential for protection against systemic mycosis, including paracoccidioidomycosis . We analyzed the reactivity of alpha beta and gamma delta T cells from unexposed Caucasian donors to P . brasiliensis yeast form components . Our results indicate: (i) alpha beta and gamma delta T cells proliferate after in vitro stimulation with lysates of P . brasiliensis; (ii) similar numbers of alpha beta T cells (f = 1/21,000) and of gamma delta T cells (f = 1/8000) respond to P . brasiliensis; (iii) P . brasiliensis-reactive gamma delta T cells express the V gamma 9V delta 2 TCR; (iv) the stimulatory activity of P . brasiliensis for both alpha beta and gamma delta T cells primarily resides in a high molecular weight (100 kDa) and in a low molecular weight (< 1 kDa) fraction; (v) the ligands responsible for stimulation of both alpha beta and gamma delta T cells are sensitive to proteinase treatment . We conclude that both alpha beta and gamma delta T cells from healthy individuals respond to ubiquitous protein antigens of P . brasiliensis.

Mycopathologia, 1994 Nov, 128(2), 67 - 73
Experimental paracoccidioidomycosis of hamster inoculated in the cheek pouch; Arruda MS et al.; We compared the granuloma morphology and immune response of hamsters inoculated with Paracoccidioides brasiliensis (Pb) into the cheek pouch, which lacks lymphatic drainage, and into the footpad, which is rich in lymphatics . Our objective was to better understand the modulation of Pb granuloma in an immunocompetent animal inoculated in an immunologically privileged site . The humoral immune response (ELISA) and cell mediated immunity (footpad test) became positive on days 7 and 14, respectively in animals inoculated into footpad and on days 35 and 60 in animals inoculated into the pouch . Typical epithelioid granulomas were observed at both sites on day 14 . The number of fungi gradually decreased from the beginning of the experiment in footpad lesions, but only after day 35 in pouch granulomas, when cell mediated immunity was detectable . The results indicate that typical epithelioid paracoccidioidomycotic granulomas may develop in the absence of a detectable immune response; however, they are incapable of controlling fungal reproduction . Lack of lymphatic drainage delays the appearance of a detectable immune response, but with time fungi escape from the pouch, elicit an immune response and reach other organs . Our results further indicate the importance of the lymphatics in the pathogenesis of paracoccidioidomycosis.

J Clin Microbiol, 1994 Oct, 32(10), 2377 - 81
Monoclonal antibody capture enzyme immunoassay for detection of Paracoccidioides brasiliensis antibodies in paracoccidioidomycosis; Camargo ZP et al.; Four murine monoclonal antibodies (MAbs 17C, 21A, 21F, and 32B) raised against the 43-kDa glycoprotein of Paracoccidioides brasiliensis were tested in a capture enzyme immunoassay (EIA) for the detection of specific human anti-gp43 immunoglobulin G in patients with paracoccidioidomycosis (PCM) . All MAbs reacted similarly in the assay . These MAbs, which detected anti-gp43 at levels of as low as 500 pg/ml, were demonstrated to specifically recognize at least two different epitopes in gp43 binding assays . Specific antibodies in the sera of patients with active PCM were detected at dilutions of as high as 1:819,200, and the reactivities of patient sera, as measured by optical densities, were found to be significantly higher than those of control sera . The comparison between classical ELISA and our capture enzyme immunoassay showed that both sensitivity and specificity were greatly improved by the latter . These MAbs represent the first specific reagents to P . brasiliensis described for use in serological tests for PCM.

J Am Acad Dermatol, 1994 Sep, 31(3 Pt 2), S91 - 102
Treatment of tropical mycoses; Restrepo A; Several subcutaneous and deep-seated mycoses are either observed more frequently in the tropical areas or are restricted to certain regions within the tropics . These mycoses include sporotichosis, chromoblastomycosis, entomophthoromycosis, eumycetoma, lobomycosis, and paracoccidioidomycosis . In sporotrichosis and paracoccidioidomycosis, therapy often results in either complete resolution or marked improvement . For decades sporotrichosis has been treated successfully with potassium iodide, but recently the triazole compounds, especially itraconazole, have proved effective and free of major side effects . The usual therapy for paracoccidioidomycosis is sulfonamides or amphotericin B; the former requires prolonged treatment, whereas the latter causes a significant degree of toxicity . Various azole derivatives (ketoconazole, fluconazole, saperconazole, and itraconazole) allow shorter treatment courses, can be given orally, and are more effective . Presently, itraconazole is the drug of choice . Chromoblastomycosis is a difficult condition to treat, especially if it is caused by Fonsecaea pedrosoi . Several therapeutic approaches have been used, including heat, surgery, cryotherapy, thiabendazole, amphotericin B combined with flucytosine, and azole derivatives, but their success has been modest . A 65% response rate has been obtained with itraconazole given for periods of 6 to 19 months; in limited trials, saperconazole appears to be more effective and requires shorter treatment courses . Only a few patients with eumycetoma respond to therapy; 70% of patients with Madurella mycetomatis respond to prolonged treatment with ketoconazole . Griseofulvin has been tried in nonresponders with partial success . Limited data in patients with Fusarium species eumycetoma indicate good responses to itraconazole . Eumycetoma caused by Pseudallescheria boydii or Acremonium species has been refractory to therapy . Therapy of entomophthoromycosis is also difficult because the diagnosis is usually established late and not all patients respond to therapy; this situation applies to infection caused by either Basidiobolus haptosporus or Conidiobolus coronatus . Although there is no consensus, African physicians prefer to use potassium iodide or trimethoprim-sulfamethoxazole . Isolated reports indicate that the azole derivatives, including the triazoles, may be effective . As for lobomycosis, all attempts at medical treatment have failed . Surgery is successful only when the lesion is small and can be fully resected; repeated cryotherapy appears to be more successful.

Mycopathologia, 1994 Sep, 127(3), 139 - 44
Neuroparacoccidioidomycosis: case reports and review; Pla MP et al.; Cerebral and cerebellar masses occurred in patients with paracoccidioidomycosis . Correct diagnosis was delayed due to overlooking the abnormal lung roentgenograms and the history of previous disease in a different localization . The fungus was identified through biopsy and direct examination of the samples . In two patients necropsy confirmed the diagnosis . None of the patients responded to amphotericin B or cotrimoxazole . A 10 year English and Latin American literature review on neuroparacoccidioidomycosis was performed through a MEDLINE and LILACS (Latin American Literature Search System) data base systems.

Braz J Med Biol Res, 1994 Sep, 27(9), 2309 - 13
Characterization of glycoprotein gp43, the major laminin-binding protein of Paracoccidioides brasiliensis; Lopes JD et al.; We have demonstrated that laminin mediates the adhesion of P . brasiliensis to monolayers of epithelial cells through specific binding to the surface glycoprotein gp43 . This binding seems to be related to the fungal pathogenesis . We now report the confirmation of these findings by scanning electron microscopy and show that some isolates that do not secrete gp43 do express the protein as seen by studying whole cell extracts . These results confirm the ability of these strains to produce paracoccidioidomycosis but should not be used for serological purposes since the absence of gp43 in exoantigens may lead to false negative results.

Braz J Med Biol Res, 1994 Sep, 27(9), 2301 - 8
Fibrosis patterns of lesions developed by athymic and euthymic mice infected with Paracoccidioides brasiliensis; Lenzi HL et al.; Athymic and euthymic mice with BALB/c background were used to study the patterns of fibrosis during ip infection with a virulent isolate of Paracoccidioides brasiliensis . Specimens from various organs were collected from the animals at 1, 4 and 7 weeks after infection and observed under light microscopy using various histologic staining methods . Lesions from the first week of infection, in both animal groups, presented a predominance of collagen III over I, carboxylated proteoglycans, and a tendency to encapsulation . From 4 weeks onward, the lesions of nu/+ mice tended to involute to macrophage-pseudoxanthomatous aggregates or to encapsulation with an increase of collagen I and sulfated proteoglycans . On the contrary, with the evolution of the infection, the nu/nu mice displayed permanently active lesions, rich in reticular fibers and carboxylated proteoglycans, with varied amounts of collagens III and I, without or with minimal encapsulation . However, independent of the type of mice, or of the type of lesions, the minimal P . brasiliensis-ECM unit was formed by a fibrillar cocoon of reticular fibers that encloses an individual yeast or a "family" composed of a mother cell plus one or various peripheral daughter cells, alone or engulfed by macrophages or giant cells . The overall difference of the lesions of nude and normal mice was not in isolated aspects of their components, but in the general architecture of the lesions . Those of nu/+ mice were either of involutive or of encapsulated type (slightly active), and those of nu/nu mice were of the sustained-expansive type (very active), without or with minimal encapsulation.

Mycoses, 1994 Sep-Oct, 37(9-10), 337 - 41
Structural alterations in Candida albicans by caffeine and caffeine salts; Mittag H; Cytological alterations of Candida albicans following exposure to subinhibitory concentrations of caffeine and caffeine salts were examined by light and transmission electron microscopy (TEM) . The effect was different from that induced by ketoconazole . Caffeine and especially caffeine salts caused an increase in unusual modes of proliferation with signs of multiple budding in Candida albicans . Structurally, such yeast cells present some resemblance to Paracoccidioides brasiliensis.

Mycoses, 1994 Sep-Oct, 37(9-10), 317 - 23
Paracoccidioides brasiliensis protoplast production by enzymatic treatment; Borba Cde M et al.; The action of the enzymes novozym 234, chitinase and zymolyase 20T on the yeast-like cells of Paracoccidioides brasiliensis was studied in an attempt to obtain protoplast release . Three enzyme systems were used: the first consisted of novozym 234 and chitinase plus 0.2 M phosphate buffer, 0.9 M sorbitol and 0.5 M sodium thioglycolate; the second consisted of novozym 234, chitinase, zymolyase 20T, buffer and osmotic stabilizer, with no sodium thioglycolate; the third consisted of the same enzymes as used in the second system but at twice the concentration, plus buffer and osmotic stabilizer . Protoplasts were only released from 72-h-old cells cultured on solid peptone-yeast extract-glucose medium (PYG) treated with the third enzyme system . Sodium thioglycolate used as pretreatment favoured protoplast release but had no such action when added to the enzyme solution, possibly by altering the activity of the enzymes, novozym 234 in particular . The osmotic stabilizer used, 0.9 M sorbitol, was probably one of the factors, in addition to the enzymes, responsible for the cytoplasmic changes observed by transmission electron microscopy in yeast phase cells and in their protoplasts.

FEBS Lett, 1994 Aug 1, 349(2), 286 - 8
The cupredoxin fold is found in the soluble CuA and CyoA domains of two terminal oxidases; Wittung P et al.; The CD spectra of the CuA domain from subunit II of Paracoccus cytochrome c oxidase and the CyoA domain of subunit II from E . coli quinol oxidase have been recorded in the wavelength region 260-185 nm . A computer program based on a set of CD spectra of proteins with known structures, and employing the statistical method of variable selection, has been used to estimate the distribution of five forms of secondary structure . The analysis was improved by including the CD spectra of azurin and plastocyanin in the basis set . For the CuA domain, an estimate from the primary structure was also made . The results show that the soluble domains have the cupredoxin fold, with very little helical structure and a predominance of beta-strands . The CyoA domain is very similar to azurin, but the beta-structure in the CuA protein resembles that in plastocyanin.

Infect Immun, 1994 Aug, 62(8), 3543 - 6
Altered expression of surface alpha-1,3-glucan in genetically related strains of Blastomyces dermatitidis that differ in virulence; Hogan LH et al.; Recent studies of the dimorphic fungal pathogens Histoplasma capsulatum and Paracoccidioides brasiliensis have suggested a role in virulence for the cell surface carbohydrate alpha-(1,3)-glucan . To investigate a possible basis for alpha-(1,3)-glucan in the pathogenicity and virulence of the dimorphic fungus Blastomyces dermatitidis, we examined three genetically related strains of B . dermatitidis that differ in their virulence for mice: wild-type virulent strain ATCC 26199; mutant strain ATCC 60915, which is 10,000-fold reduced in virulence; and mutant strain ATCC 60916, which is avirulent . Immunologic quantitation of cell wall alpha-(1,3)-glucan revealed that the mutant yeasts were almost devoid of this sugar moiety, in contrast to the high concentration of alpha-(1,3)-glucan on the cell wall of the wild-type yeasts . These differences are discussed in relation to previous studies of yeast surface expression of the WI-1 antigen and recognition and binding of the related strains by human monocyte-derived macrophages.

Mycopathologia, 1994 Aug, 127(2), 89 - 93
Systematic evaluation of the adrenocortical function in patients with paracoccidioidomycosis; Colombo AL et al.; Limited cortisol response to ACTH stimulation has been documented in 22 to 48% of patients with paracoccidioidomycosis (PM) . Different approaches to interpret the test and inadequate selection of patients preclude an accurate appraisal of the actual incidence of adrenal insufficiency in PM . Rapid cosyntropin (ACTH) stimulation tests were performed in 38 consecutive patients (9 with the localized and 29 with the disseminated form of PM) and 40 normal controls . Subnormal cortisol responses to ACTH (60 minutes post-ACTH values below 455 nmol/l, 95% confidence limits) were found in only 4 patients (14%) with disseminated PM . If a retrospective sample of 6 patients studied previously (in whom tests were indicated due to clinical suspicion of Addison's disease) were included, or if the absolute cortisol increment above baseline was used for interpretation, we would find figures closer to those previously reported (23 and 24%, respectively) . These data reflect that non-systematic evaluation or selection of a substandard criterion to interpret the test overestimates the frequency of adrenocortical insufficiency in PM.

Mycopathologia, 1994 Aug, 127(2), 69 - 71
Alkaline phosphatase at the cell wall of the yeast phase of Paracoccidioides brasiliensis; Campo-Aasen I et al.; The activity of alkaline phosphatase demonstrated by histochemical techniques was shown at the cell wall of the yeast form of Paracoccidioides brasiliensis at 3, 6, and 9 days of culture . The results showed a very active deposition at the cell wall as early as 9 days of culture of the fungus which made us think an inactive salt precipitate was also present.

Clin Diagn Lab Immunol, 1994 Jul, 1(4), 390 - 3
Effects of histoplasmin M antigen chemical and enzymatic deglycosylation on cross-reactivity in the enzyme-linked immunoelectrotransfer blot method; Zancope-Oliveira RM et al.; The enzyme-linked immunoelectrotransfer blot (EITB) method was evaluated as a suitable method for detecting antibodies against M antigen of Histoplasma capsulatum by use of both glycosylated and deglycosylated M protein of histoplasmin (HMIN) . Sera from patients with histoplasmosis, paracoccidioidomycosis, blastomycosis, coccidioidomycosis, and aspergillosis were tested by the EITB with glycosylated M protein of HMIN . This assay demonstrated 100% sensitivity with histoplasmosis serum samples, all of which reacted with the 94-kDa glycoprotein (M antigen) . Although the EITB is highly sensitive, it is not specific for histoplasmosis when glycosylated M protein is used as an antigen . A total of 81% of paracoccidioidomycosis, 25% of blastomycosis, 33% of coccidioidomycosis, 73% of aspergillosis, and 16% of tuberculosis serum samples cross-reacted with M protein of HMIN and yielded patterns indistinguishable from those obtained with histoplasmosis serum samples . The EITB reactions with both untreated M antigen and M antigen altered by periodate oxidation or by deglycosylation with endoglycosidases were compared . Cross-reactions with heterologous sera in the EITB could be attributed to periodate-sensitive carbohydrate epitopes, as reflected by the increase in the test specificity from 46.1 to 91.2% after periodate treatment of M protein . The EITB for the detection of antibodies to M antigen is a potential diagnostic test for histoplasmosis, provided that periodate-treated M protein is used as an antigen.

Clin Exp Immunol, 1994 Jul, 97(1), 113 - 9
Pathogenicity and immunogenicity of Paracoccidioides brasiliensis isolates in the human disease and in an experimental murine model; Singer-Vermes LM et al.; The pathogenicity and immunogenicity of six recently isolated Paracoccidioides brasiliensis samples derived from patients presenting distinct and well defined clinical forms of paracoccidioidomycosis (PCM) were compared as to their virulence, tropism to different organs and ability to induce specific cellular and humoral immune response in susceptible (B10.A) inbred mice . Isolates Pb44 and Pb47 were obtained from acute cases, Pb50 from a chronic severe form, Pb45 from a chronic moderate case and both Pb56 and Pb57 from chronic mild forms of PCM . Pathogenicity and tropism of each fungal sample were evaluated by LD50% estimation, examination of gross lesions on various organs at 2, 4, 12 and 16 weeks post-infection, and by colony-forming unit (CFU) counts in the lungs at week 16 post-infection of mice . Fungal tropism in human PCM and in B10.A mice was always dissociated . A well defined relationship between virulence of the fungal sample and the clinical findings of the correspondent patient was not evident, although a tendency to higher LD50% and less intense paracoccidioidic lesions was observed in mice infected with Pb56 and Pb57 . The specific DTH response patterns varied according to the infectant sample, but positive DTH reactions at the beginning of the infection and a tendency to anergy or low DTH responses at week 12 and/or week 16 post-infection were always observed . A correspondence between the DTH response in humans and in mice was noticeable only when the isolates from the most benign cases (Pb56 and Pb57) were considered . The specific antibody patterns in mice and in the correspondent patients were also not analogous . Collectively, these results indicate that an association between the fungal pathogenicity and immunogenicity in the human disease and in susceptible mice was discernible only when isolates obtained from very mild cases (Pb56 and Pb57) were considered.

Pediatr Infect Dis J, 1994 Jun, 13(6), 510 - 5
Severe acute paracoccidioidomycosis in children; Benard G et al.; This report describes clinical and immunologic features of five illustrative cases of paracoccidioidomycosis in previously healthy children . All had disseminated disease and two of them died despite treatment . The major clinical presentation in four patients was fever and diffuse superficial and intraabdominal adenopathy, with or without hepatosplenomegaly . Other sites were also affected: three patients had multiple osteoarticular lesions, occasionally with intense tissue destruction; two had cutaneous eruptions; two had pericardial effusions; and two had pulmonary involvement, once considered an organ spared in the young . We detected variable lymphocyte responses to mitogens and to Candida albicans antigen and non-responsiveness to Paracoccidioides brasiliensis cell wall antigen . High concentrations of serum immunoglobulins and anti-P . brasiliensis antibodies were present . These immune alterations tended to resolve with treatment, suggesting a reversible nature of the immune defect . We conclude that this mycosis has a high morbidity and mortality in children, which is probably related to an antigen-specific immunodeficiency . Further studies are needed to increase knowledge of this mycosis in children.

J Clin Microbiol, 1994 Jun, 32(6), 1566 - 74
Immunohistochemical detection of a novel 22- to 25-kilodalton glycoprotein of Paracoccidioides brasiliensis in biopsy material and partial characterization by using species-specific monoclonal antibodies; Figueroa JI et al.; Two murine monoclonal antibodies (MAbs) specific to Paracoccidioides brasiliensis (as determined by enzyme-linked immunosorbent assay {ELISA} and Western blot {immunoblot}) were produced by using a modification of standard hybridization protocols, with cyclophosphamide included as an immunomodulator to abolish responses to highly cross-reactive immunodominant epitopes . MAbs PS14 and PS15 are two different clones which exhibit similar characteristics by ELISA and Western blot . They are directed against a 22- to 25-kDa antigen which is present in P . brasiliensis and which could not be identified in other dimorphic fungi by ELISA or Western blot . Partial purification of the antigen was accomplished by isoelectric focusing, and deglycosylation studies suggested that the 22- to 25-kDa antigen is a glycoprotein with a pI of between 4.5 and 5 and that O-linked sugars may be part of the recognized epitope . The MAbs stained the cytoplasm of P . brasiliensis yeast and hyphal cells in cryostat sections of fresh cultures of the fungus . In addition, the MAbs stained the wall of paracoccidioidomycotic granulomas, as well as the cytoplasm of the fungus, as determined by the use of immunofluorescence, immunoperoxidase, and immuno-alkaline phosphatase staining techniques in paraffin-embedded sections of human biopsy material, and they failed to stain granulomas resulting from other clinical conditions . These findings suggest that these MAbs have potential use in the immunohistochemical identification of P . brasiliensis.

Mycopathologia, 1994 Jun, 126(3), 137 - 46
Delayed-type hypersensitivity response in an isogenic murine model of paracoccidioidomycosis; Fazioli Rdos A et al.; The specific delayed-type hypersensitivity (DTH) response was evaluated in resistant (A/SN) and susceptible (B10.A) mice intraperitoneally infected with yeasts from a virulent (Pb18) or from a non-virulent (Pb265) Paracoccidioides brasiliensis isolates . Both strains of mice were footpad challenged with homologous antigens . Pb18 infected A/SN mice developed an evident and persistent DTH response late in the course of the disease (90th day on) whereas B10.A animals mounted a discrete and ephemeral DTH response at the 14th day post-infection . A/SN mice infected with Pb265 developed cellular immune responses whereas B10.A mice were almost always anergic . Histological analysis of the footpads of infected mice at 48 hours after challenge showed a mixed infiltrate consisting of predominantly mononuclear cells . Previous infection of resistant and susceptible mice with Pb18 did not alter their DTH responses against heterologous unrelated antigens (sheep red blood cells and dinitrofluorobenzene) indicating that the observed cellular anergy was antigen-specific . When fungal related antigens (candidin and histoplasmin) were tested in resistant mice, absence of cross-reactivity was noted . Thus, specific DTH responses against P . brasiliensis depend on both the host's genetically determined resistance and the virulence of the fungal isolate.

Mycoses, 1994 Jun-Jul, 37(5-6), 165 - 9
Viability and morphological alterations of Paracoccidioides brasiliensis strains preserved under mineral oil for long periods of time; da Silva AM et al.; We evaluated the survival and the morphological alteration of 70 strains of Paracoccidioides brasiliensis maintained in the Fungal Culture Collection of Institute Oswaldo Cruz and initially preserved by successive subculturing and later under mineral oil at room temperature from 1923 to 1992 . Of the 70 strains preserved under mineral oil, 18 (26%) continued to be viable . The mycelia of the 18 viable strains and the mycelia of four representatives of the non-viable strains presented transitional micromorphology under mineral oil . Macroscopy and microscopy of the first subcultures of five of the seven strains, taken as random samples from the 18 viable strains and plated onto peptone-yeast extract-glucose agar, glucose-glycine-yeast extract agar and brain-heart infusion agar media at room temperature, revealed that all samples appeared to be in a transition phase (Y<-->M) . These five strains were unable to grow and complete thermal dimorphism at a temperature of 37 degrees C . Only two strains were able to complete the entire dimorphic process . The period of preservation of these strains under oil was relatively short, i.e . 9 and 10 years . The results demonstrate that less widely spaced subculturing and more appropriate culture and environmental conditions are needed to preserve P . brasiliensis strains under oil.

Microbiology, 1994 May, 140 ( Pt 5), 1189 - 94
Involvement of cell wall glucans in the genesis and persistence of the inflammatory reaction caused by the fungus Paracoccidioides brasiliensis; Silva CL et al.; The role of cell wall polysaccharides in leucocyte recruitment and granuloma formation in paracoccidioidomycosis was investigated . The inflammatory cells recruitment to the peritoneal cavity in rats inoculated with cell wall fraction (CW-265 or F1-265) from an avirulent strain of Paracoccidioides brasiliensis (Pb265), was greater than that observed for the cell wall fraction (CW-HC or F1-HC) recovered from the virulent strain (PbHC) . Moreover, the inoculation of F1-HC and F1-265 into the subcutaneous layer of mice resulted in the formation of nodular and not progressive granulomatous lesions . The size and mean time of evolution of these lesions was proportional to the degree of virulence of the sample from which they were derived . Analyses showed that both F1 fractions contained beta-glucan and chitin . Only beta-glucan was able to trigger attraction and concentric organization of polymorphonuclear neutrophils and macrophages at the inflammatory foci, and the difference in the concentration of this compound in the cell walls of PbHC and Pb265 could explain the inflammatory capacity exhibited by the two strains of P . brasiliensis.

Rev Inst Med Trop Sao Paulo, 1994 May-Jun, 36(3), 217 - 23
Experimental paracoccidioidomycosis in hamster: transmission electron microscopy of inoculation site lesion; Coelho KI et al.; Interaction between Paracoccidioides brasiliensis (Pb) and inflammatory cells in hamster testis was studied sequentially by transmission electron microscopy . In early lesions (six hours after inoculation), polymorphonuclear neutrophils (PMNs) were the major and mononuclear cells and eosinophils were the minor constituents of the inflammatory cells . PMNs were later replaced by mononuclear cells . Viable Pb cells were phagocytosed or surrounded by inflammatory cells . Preserved Pb cells usually had broad host-parasite interphases, whereas dying ones had narrow interphases . The outer layer of the fungus wall was sometimes broken by PMN in some focal points, broken pieces being peeled off and phagocytosed . Small Pb cells were uninuclear, and were often related to broad interphase . Large Pb cells were multinucleated with irregularly shaped wall, and sometimes had lomasome and/or myelin like structures . Different interaction patterns of Pb with inflammatory cells may be due to functionally different host cell flow to the inoculation site or due to the age of Pb cells or both.

Infect Immun, 1994 Apr, 62(4), 1494 - 6
Inhibitory effect of deferoxamine or macrophage activation on transformation of Paracoccidioides brasiliensis conidia ingested by macrophages: reversal by holotransferrin; Cano LE et al.; Conidia of P . brasiliensis ingested by murine macrophages at 37 degrees C showed enhanced transformation to yeast cells and further intracellular growth compared with conidia in culture medium alone . Treatment of macrophages with the iron chelator deferoxamine inhibited the intracellular conidium-to-yeast transformation . Cytokine-activated macrophages could also exert this inhibitory effect . Holotransferrin reversed the inhibitory effect of either deferoxamine or activated macrophages on intracellular conidium-to-yeast transformation . These results indicate that iron restriction is one of the mechanisms by which activated macrophages control the intracellular transformation of ingested conidia and growth of yeast cells.

Infect Immun, 1994 Apr, 62(4), 1465 - 9
Binding of Paracoccidioides brasiliensis to laminin through surface glycoprotein gp43 leads to enhancement of fungal pathogenesis; Vicentini AP et al.; Extracellular matrix protein laminin binds specifically to yeast forms of Paracoccidioides brasiliensis and enhances adhesion of the fungus to the surface of epithelial Madin-Darby canine kidney cells in vitro . Immunoblotting of fungal extracts showed that the gp43 glycoprotein is responsible for adhesion . This was confirmed by binding assays using purified gp43, with a Kd of 3.7 nM . The coating of P . brasiliensis yeast forms with laminin before injection into hamster testicles enhanced the fungus virulence, resulting in a faster and more severe granulomatous disease . These results indicate that interaction of fungi with extracellular matrix elements may constitute a basis for the evolution of fungal infection toward regional spreading and dissemination.

Ann Trop Med Parasitol, 1994 Apr, 88(2), 197 - 207
Paracoccidioidin and histoplasmin sensitivity in Tupí-Mondé Amerindian populations from Brazilian Amazonia; Coimbra Junior CE et al.; A cross-sectional epidemiological survey for paracoccidioidomycosis and histoplasmosis, including skin tests with paracoccidioidin and histoplasmin, physical examinations and X-rays, was conducted among three Tupi-Monde Amerindian populations from Brazilian Amazonia . The study followed the diagnosis of an increasing number of cases of paracoccidioidomycosis among the Surui in recent years . Positivity rates to paracoccidioidin and histoplasmin (> or = 5 mm of intradermal induration 24-48 h post-injection) were 43.8% and 78.7% for the Surui, 6.4% and 5.8% for the Gaviao and 14.9% and 80.5% for the Zoro, respectively . There was no significant difference in the results for males and females but marked differences were noted across age groups . The results of the univariate analysis were confirmed after adjustment for confounding variables by multiple logistic regression analysis: paracoccidioidin positivity was relatively high in the Surui and histoplasmin positivity was relatively high in the Surui and Zoro . The Surui's greater exposure to Paracoccidioides brasiliensis, the causative agent of paracoccidioidomycosis, is probably associated with their adoption of new subsistence practices . The epidemiology of this mycosis among the Tupi-Monde appears to be related to the environmental and socio-economic changes taking place in Amazonia.

J Clin Microbiol, 1994 Apr, 32(4), 867 - 70
Comparative evaluation of chemiluminescent DNA probe assays and exoantigen tests for rapid identification of Blastomyces dermatitidis and Coccidioides immitis; Padhye AA et al.; Chemiluminescent DNA probe (Accuprobe) assays developed by Gen-Probe, Inc . (San Diego, Calif.), for the rapid identification of Blastomyces dermatitidis and Coccidioides immitis were evaluated and compared with the exoantigen test by using 74 mycelial cultures of B . dermatitidis and 72 mycelial cultures of C . immitis . Seventeen isolates of the dimorphic pathogen Paracoccidioides brasiliensis were included because of their gross morphologic and antigenic relatedness to B . dermatitidis . The heterologous fungi, namely, species of Chrysosporium, which are often confused with B . dermatitidis, and species of Malbranchea, which morphologically resemble C . immitis, were tested . All 74 of the B . dermatitidis mycelial isolates were correctly identified by the Accuprobe assay for B . dermatitidis within 2 h . However, the B . dermatitidis probe cross-hybridized with rRNA extracts of 10 of the 17 P . brasiliensis isolates, misidentifying them as B . dermatitidis . All 72 of the C . immitis isolates were identified correctly with the C . immitis probe . None of the other heterologous fungi belonging to Chrysosporium spp., Malbranchea spp., Onychocola canadensis, and Geotrichum sp . were cross-reactive with the B . dermatitidis and C . immitis probes . The exoantigen tests specifically identified 74 B . dermatitidis, 72 C . immitis, and 17 P . brasiliensis isolates within 48 to 72 h and differentiated the related heterologous fungi from the three dimorphic fungal pathogens.

J Med Microbiol, 1994 Mar, 40(3), 159 - 64
Support of Paracoccidioides brasiliensis multiplication by human monocytes or macrophages: inhibition by activated phagocytes; Moscardi-Bacchi M et al.; The interaction of human monocytes or monocyte-derived macrophages and yeast-form Paracoccidioides brasiliensis was studied in vitro . Yeast cells were readily ingested by adherent monocytes or macrophages . Multiplication of P . brasiliensis, measured by growth as colony forming units (cfu) on a supplemented medium with good plating efficiency, was greater in monocyte co-cultures compared to the number of cfu obtained from complete tissue-culture medium (CTCM) . Multiplication increased with time in macrophage co-cultures, e.g., from two-six-fold in 24 h to nine-fold in 72 h . Microscopic observations indicated that ingested yeast cells multiplied inside macrophages . When monocytes were treated with supernate cytokines (CK) from concanavalin-A-stimulated mononuclear cells, then co-cultured with P . brasiliensis, multiplication was significantly inhibited compared with control monocyte co-cultures . Treatment of macrophages--derived from monocytes by culture in vitro for 3 days--for a further 3 days with CK resulted in maximal inhibition of multiplication over the subsequent 72 h . Similarly, when monocyte-derived macrophages (after culture for 7 days) were treated for 3 days with recombinant human gamma-interferon (IFN; 300 U/ml) or CK they restricted multiplication of P . brasiliensis by 65% and 95%, respectively, compared with control macrophages . Antibody to IFN abrogated the effect of IFN or CK treatment . These findings show that ingested P . brasiliensis can multiply in human monocytes or macrophages and that this multiplication can be restricted by activated monocytes or macrophages.

Arq Neuropsiquiatr, 1994 Mar, 52(1), 82 - 6
{Paracoccidioidomycosis evidencing spinal cord involvement treated with success by fluconazole}; de Moura LP et al.; The involvement of central nervous system in paracoccidiodomycosis has rarely been described, with an incidence rate varying from 9.99% to 27.27% . There are two basic forms of clinical presentation: meningeal and tumor-like (abscesses, granulomas, nodules, and cysts) . The Paracoccidioides brasiliensis is preferentially described in cerebral hemispheres, cerebellum, medulla oblonga and meninges, and exceptionally in the spinal cord . The authors present a case of paracoccidioidomycosis which diagnosis was achieved by microscopic examination of material from oral lesions and specific serology . The patient presented clinical signs of spinal cord involvement confirmed by lesions found in magnetic resonance imaging . They emphasize the inedit therapeutic response to a new antifungal agent (fluconazole) used for the first time in this kind of clinical manifestation, and the excellent prognosis when diagnosis is promptly made.

J Investig Allergol Clin Immunol, 1994 Mar-Apr, 4(2), 91 - 5
Classical and alternative complement pathway activation in paracoccidioidomycosis; de Messias IT et al.; The following study presents evidence of complement activation in paracoccidioidomycosis (PCM) . Twenty-eight untreated patients were studied from endemic areas of Parana in southern Brazil . The activation of the classical pathway, evaluated by the C4d/C4 ratio, was significantly elevated in the patients compared to the control population (p < 0.005) . Six patients were examined prospectively with the C4d/C4 assay during treatment and they showed a decrease in this ratio associated with clinical improvement . The activation of the alternative pathway was determined by rocket immunoelectrophoresis of fragment Ba . These levels were also significantly higher in the patient group in comparison to the controls (p < 0.0005) . The prospective study also showed a significant variation in the Ba levels associated with clinical improvement (p < 0.01) . Furthermore, the levels of C3, C4, CH50 and anti-Paracoccidioides brasiliensis IgG were determined in all patients . The anti-P . brasiliensis IgG levels showed a weak positive correlation with the C4d/C4 ratio (r{S} = 0.45; p < 0.03) . The C3, C4 and CH50 levels did not show significant variations from the normal ranges . Our results suggest the involvement of both complement pathways, classical and alternative, in PCM and their association with disease activity.

FEBS Lett, 1994 Feb 14, 339(1-2), 142 - 6
Studies on the proton-translocating NADH:ubiquinone oxidoreductases of mitochondria and Escherichia coli using the inhibitor 1,10-phenanthroline; Finel M et al.; Mitochondrial NADH:ubiquinone oxidoreductase (complex I) is uncompetitively inhibited by 1,10-phenanthroline (OP) . EPR spectroscopy of submitochondrial particles indicates that OP, similarly to rotenone, inhibits electron transfer between the Fe-S clusters of complex I and the ubiquinone pool . The proton-translocating NADH dehydrogenase (NDH1) of E . coli is more sensitive to OP than is NDH1 of Paracoccus . EPR spectroscopy of membranous E . coli NDH1 shows that two slow- and one fast-relaxing Fe-S clusters become detectable upon reduction by NADH in the presence of OP . However, none of them resembles the mitochondrial cluster 2.

J Oral Pathol Med, 1994 Feb, 23(2), 85 - 7
Paracoccidioidomycosis manifesting as oral lesions: clinical, cytological and serological investigation; Sposto MR et al.; Paracoccidioidomycosis (South American blastomycosis) is a systemic mycosis which can be associated with oral lesions . This study on a group of 14 patients showed oral lesions mainly on the gingival or alveolar mucosa, with pulmonary involvement detectable on chest radiography in most . Microscopic detection of the fungus on a direct smear showed positive results in all 14 patients . Serological investigations including immunodiffusion, counterimmunoelectrophoresis and immunoblot were also positive in 100% of cases . The results suggest that direct smear together with serology may obviate the need for lesional biopsy for the diagnosis of oral paracoccidioidomycosis.

J Med Microbiol, 1994 Feb, 40(2), 124 - 8
Synthesis of heat-shock proteins in mycelia and yeast forms of Paracoccidioides brasiliensis; Goldani LZ et al.; The induction of heat-shock proteins has been postulated to play a role not only in thermo-adaptation, but also in phase transition of the dimorphic fungi . In this study, we used yeast and mycelial forms of the thermally dimorphic fungus Paracoccidioides brasiliensis to evaluate the effect of temperature on the induction of the heat-shock response . We also evaluated protein synthesis by P . brasiliensis caused by exposure to low pH and H2O2 . Analysis of protein synthesis by SDS-PAGE disclosed that P . brasiliensis mycelia increased synthesis of all major constitutive proteins when stressed at 37 degrees C and increased synthesis of three non-constitutive proteins of 134, 82 and 28 kDa at 40 degrees C . Yeasts incubated at 40 degrees C showed decreased synthesis of five constitutive proteins (136, 98, 62, 57 and 54 kDa) and the appearance of three new proteins (134, 82 and 28 kDa) . There was a decrease in the synthesis of all major constitutive proteins except for three proteins of 141, 136 and 16 kDa when yeast cells were incubated at 25 degrees C . When stressed by low pH and H2O2, P . brasiliensis yeast increased synthesis of one (134 kDa) and five (134, 104, 82, 52 and 40 kDa) non-constitutive proteins, respectively . P . brasiliensis mycelia and yeast forms disclosed the same profile of protein synthesis when stressed at temperatures that trigger phase transition (37 degrees C for mycelia; 25 degrees C for yeast).(ABSTRACT TRUNCATED AT 250 WORDS)

J Med Vet Mycol, 1994, 32(1), 65 - 9
Gelatinase activity of exoantigens from virulent and non-virulent isolates of Paracoccidioides brasiliensis; Vaz CA et al.; Differences in the occurrence of components with gelatinase activity were detected among four isolates of Paracoccidioides brasiliensis: Pb339 and Pb18 (highly virulent), and Pb265 and Pb18AV (very low virulence) . Culture filtrates from these isolates were electrophoresed in substrate gels and tested for gelatinase activity . Pb339 showed three enzyme bands of apparent molecular masses: 43, 53 and 78 kDa; Pb18 had two bands, one at 59 kDa and another with molecular mass higher than 78 kDa . Isolate Pb18AV showed only one band at 78 kDa and Pb265 exhibited a component of molecular mass which failed to enter the separating gel.

Rev Soc Bras Med Trop, 1994 Jan-Mar, 27(1), 11 - 4
{The evaluation of ketoconazole in mice inoculated with Paracoccidioides brasiliensis by liver and spleen histopathology and by the intradermal paracoccidioidin reaction}; Silva MR et al.; Male albino mice were inoculated intravenously with 0.5 x 10(7) viable yeast forms of P . brasiliensis (strain 2052) . These animals were treated with two doses of ketoconazole (50 and 100 mg/kg) during fifty days and the sacrificed . We studied the presence of P . brasiliensis, the inflammatory granulomatous response of liver and spleen and the anti P . brasiliensis delayed hypersensitivity response measured by the footpad test after 48 hours . It was observed that: 1 . animals infected and treated with ketoconazole showed reduction in the number of fungi in the organs studied; 2 . there was no difference in the number of granulomas among animals treated and non-treated; 3 . the cutaneous delayed tests intensity was similar in all animals.

J Med Vet Mycol, 1994, 32(3), 199 - 203
Chromosome aberrations in lymphocyte cultures from paracoccidioidomycosis patients; Freire-Maia DV et al.; Blood cell lymphocyte chromosomes from untreated (UT) and clinically-cured (CC) patients with paracoccidioidomycosis and from healthy (control) people (CO) were studied . The frequency of aneuploid cells in the UT patients was higher than in the CC and CO individuals . The frequency of metaphase cells with premature centromere division was significantly higher in the UT than in the CC and CO group . No structural aberration and no statistically significant difference in the frequency of polyploidy was observed in the three groups studied . Our findings are indicative of an aneugenic (aneuploidy-inducing) action of infection by Paracoccidioides brasiliensis.

J Med Vet Mycol, 1994, 32(3), 189 - 97
Infection and apparent invasion of Vero cells by Paracoccidioides brasiliensis; Mendes-Giannini MJ et al.; Paracoccidioides brasiliensis probably uses many different mechanisms to establish itself in the host and cause disease . In this work, we assess an in vitro model system which uses cultured mammalian cells to investigate the virulence factors of P . brasiliensis . We were able to demonstrate an invasion process of the yeast form of this fungus in Vero cell cultures . We deduced that the overall invasive process involved three steps: adhesion, followed by invasion of individual epithelial cells and spread to adjacent cells.

J Clin Lab Anal, 1994, 8(6), 424 - 31
Chemiluminescent immunoassays: discrimination between the reactivities of natural and human patient antibodies with antigens from eukaryotic pathogens, Trypanosoma cruzi and Paracoccidioides brasiliensis; Almeida IC et al.; Quantitative chemiluminescent enzyme-linked immunosorbent assay (ELISA) and dot-blotting procedures were developed to evaluate the reactivity of human antibodies with crude antigens and purified molecules of parasites and fungi, mainly Trypanosoma cruzi and Paracoccidioides brasiliensis . Reproducible, highly sensitive, and strictly dose-responding results were obtained, with the specificity depending on the kind of antigen used . Mixed antigens (epimastigote membrane and HIV-1 heptapeptide) applied in dots could be independently recognized by specific sera . Purified antigens (T . cruzi F2/3 and P . brasiliensis gp43) at very small concentrations gave specific reactions with patients' sera diluted > or = 1:1,000 and were very poorly reactive or unreactive with natural antibodies using the chemiluminescent immunoassays . P . brasiliensis crude antigen Fava Netto polysaccharide antigen (FNPA) contained peptide epitopes recognized by natural antibodies and carbohydrate epitopes reactive with sera from histoplasmosis patients . It is very important that sensitive chemiluminescence immunoassays be used with purified antigenic molecules to ensure specificity for the diagnosis and follow-up of parasitic and fungal infections.

J Med Vet Mycol, 1994, 32(5), 381 - 8
Production and regeneration of protoplasts from the Y-phase of the human pathogenic fungus Paracoccidioides brasiliensis; San-Blas F et al.; Protoplasts of Y-cells and partially converted M-cells from several human isolates of Paracoccidioides brasiliensis were obtained with a combined enzyme system containing Novozym 234 and a chitinase . A laboratory made extract from Trichoderma harzianum CBS-345-33 supplemented with chitinase induced the release of protoplasts from partially converted M-cells but not from established Y-cells . A similar yield of protoplasts (1-2 x 10(5) ml-1 after 16 h) was produced by using either enzymatic system . Protoplasts regenerated on nutrient gelatin at 23 degrees C at a frequency of 0.1%.

J Med Vet Mycol, 1994, 32(5), 331 - 41
Phylogeny of dermatophytes and dimorphic fungi based on large subunit ribosomal RNA sequence comparisons; Leclerc MC et al.; The phylogeny of dermatophytes and dimorphic fungi was considered using the large-subunit of ribosomal RNA (25S rRNA) . Aligned sequences of 595 nucleotides covering the two most divergent domains D1 and D2, permitted a comparison of phylogenetic relationships at different levels . The dimorphic species (Onygenaceae) were significantly separated from dermatophytes (Arthrodermataceae) and from a third group including geophilic or very weakly pathogenic species (Onygenaceae and Gymnoascaceae) . On a species level, the varietal status of Histoplasma duboisii and Histoplasma farciminosum, as close relations of Histoplasma capsulatum, was confirmed . The dimorphic fungus Emmonsia parva, in spite of a completely different parasitic form (adiaspores instead of yeast-like cells), clustered with Blastomyces dermatitidis which has a perfect form resembling that of H . capsulatum . From our data, teleomorphs of E . parva, Paracoccidioides brasiliensis and H . farciminosum, three dimorphic fungi known only under their anamorphic states, should belong to the family of Onygenaceae and the genus Ajellomyces . Among Arthrodermataceae, and family containing the most keratinophilic species, it was not possible to establish a clear hierarchy of species . Only Ctenomyces serratus, the species adapted to degrade keratin of feathers, Trichophyton ajelloi and Trichophyton terrestre were significantly separated . The speciation of true dermatophytes resulted most likely from a very recent evolution by adaptation to parasitism . Among species used as outgroups, the two emerging pathogens Pseudallescheria boydii (Scedosporium apiospermum) and Scedosporium prolificans (Scedosporium inflatum) were shown to be closely related to each other.

J Med Vet Mycol, 1994, 32 Suppl 1, 203 - 10
Cytokines in the host response to mycotic agents; Murphy JW et al.; In summary, different approaches have been taken to understand cytokine responses to different fungal infections . Singer-Vermes and co-investigators indirectly examined cytokine responses to paracoccidioidomycosis by studying the types of cellular and humoral immune responses that were induced in resistant and susceptible mouse strains . Their results implicated Th1 cell responses in the resistant mouse strain and Th2 cell responses in the mouse strain susceptible to paracoccidioidomycosis . By measuring cytokine production and through cytokine depletion experiments, Wu-Hsieh showed that besides IFN gamma, TNF alpha was important in host defences against the intracellular pathogen, H . capsulatum . Both cytokines play important roles in the regulation of other cytokines . In histoplasmosis, the dynamics of the complex interactions amongst cytokines govern the efficiency of host clearance of the fungus from tissues . Ferrante and collaborators, examining TNF alpha and TNF alpha receptors on neutrophils presented data showing that TNF alpha plays an important role in the activation of neutrophils for anti-Candida activity . Through the detection of cytokine mRNAs with RT-PCR, Moser and co-workers found that cytokine mRNAs of macrophage origin were produced preferentially in the lungs of mice infected with Histoplasma or Blastomyces . A great challenge still lies ahead of us . It is well understood that the interactions of cytokines are extremely complex at the levels of the induction and expression of the immune responses as well as on effects on natural cellular defences . Work accomplished thus far has laid the ground work for future studies in the effort to dissect host cytokine responses and to understand the roles of cytokines in protection against fungal infections.

J Med Vet Mycol, 1994, 32 Suppl 1, 321 - 8
Fungal polysaccharides; San-Blas G et al.; Fungal polysaccharides are cell wall components which may act as antigens or as structural substrates . As antigens, the role of mannans in Saccharomyces cerevisiae and Candida albicans, and of glycoproteins in Aspergillus fumigatus are discussed . Analyses on beta-glucan synthetase in Paracoccidioides brasiliensis and the inhibitory effect of Hansenula mrakii killer toxin on beta-glucan biosynthesis are also considered.

J Med Vet Mycol, 1994, 32(1), 77 - 81
Nature of the reactive epitopes in Paracoccidioides brasiliensis polysaccharide antigen; Rodrigues EG et al.; Fava Nettos' polysaccharide antigen (FNPA), shown to detect humoral and cellular responses in paracoccidioidomycosis, was investigated . Skin tests with FNPA were negative after alkaline hydrolysis or depletion of gp43 peptide epitopes . Purified antibodies to FNPA or gp43 from paracoccidioidomycosis patients cross-reacted, showing common epitopes and FNPA-specific ones . Normal human sera, in contrast to paracoccidioidomycosis sera, were unreactive with gp43 but recognized epitopes of FNPA susceptible to alkaline hydrolysis and pronase treatment . Histoplasmosis patients sera strongly reacted with FNPA carbohydrate epitopes.

J Med Vet Mycol, 1994, 32(1), 37 - 45
Plasma amylase levels as a marker of disease severity in an isogenic murine model of paracoccidioidomycosis; Xidieh CF et al.; Survival patterns after peritoneal infection with Paracoccidioides brasiliensis vary according to the mouse strain and to the virulence of the fungal isolate . It has previously been observed that a significant increase in plasma amylase levels occurs only when susceptible mice (B10.A) were infected with a virulent isolate (Pb18) . In order to verify if increased amylase levels correlate with susceptibility to P . brasiliensis infection, 12 mouse strains with different susceptibility patterns to this fungus were investigated after infection with Pb18 . When compared with their respective controls, C57BI/6, B10D2/oSn, B10D2/nSn, C3H/HeJ, B10.A and BALB/c mice showed a conspicuous amylase increase and AKR, (NZB x NZW)F1, CBA/J, (A/Sn x B10.A)F1, A/Sn and DBA/2 absence of alteration . The influence of the infecting fungal isolate on this enzymatic parameter was investigated using B10.A mice and fungal isolates with diverse degrees of virulence . When compared with their non-infected controls, mice infected with Pb45 or Pb47 showed a very high amylase increase, with Pb44 or Pb18 a high one and with Pb50 or Pb265 a discrete increase . On the whole, there is an inverse correlation between survival times after infection and the increase in amylase levels . Thus, measurement of plasma amylase is a satisfactory parameter to evaluate the severity of paracoccidioidomycosis in mice.

Otolaryngol Clin North Am, 1993 Dec, 26(6), 1007 - 35
Fungal infections of the nose and paranasal sinuses . Part I; Blitzer A et al.; This article details the incidence, etiology, classification, and radiographic findings of fungal infections . Major infections discussed include aspergillosis, mucormycosis, blastomycosis, candidiasis, chromblastomycosis, coccidioidomycosis, histoplasmosis, paracoccidioidomycosis, and rhinosporidiosis.

Mayo Clin Proc, 1993 Dec, 68(12), 1151 - 7
Rhinoscleroma: a growing concern in the United States? Mayo Clinic experience; Andraca R et al.; Rhinoscleroma is a chronic, progressive, granulomatous infection of the upper airways caused by the bacterium Klebsiella rhinoscleromatis . Although most cases occur in developing countries, recent immigration patterns have led to an increasing number of patients with rhinoscleroma in the United States . Rhinoscleroma may mimic various inflammatory and neoplastic processes, including leprosy, paracoccidioidomycosis, sarcoidosis, basal cell carcinoma, and Wegener's granulomatosis . Current therapy consists of a combination of surgical debridement and prolonged antimicrobial therapy . Rhinoscleroma should be added to the list of opportunistic infections that can occur in patients with human immunodeficiency virus.

Mycopathologia, 1993 Dec, 124(3), 157 - 61
A comparative study of four different staining methods for estimation of live yeast form cells of Paracoccidioides brasiliensis; Sano A et al.; A comparative study of four different staining methods for estimation of live yeast form cells of Paracoccidioides brasiliensis was carried out . The staining methods used were fluorescent staining, vital dye exclusion tests with erythrosin B and by Janus green and lactophenol cotton blue staining . Colony forming units (cfu) of the yeast form of eight P . brasiliensis isolates on brain heart infusion agar (BHIA) supplemented with 4% horse serum plus 5% P . brasiliensis cell extract (BHIA + HS + EXT) were examined for reliability of staining in determining the number of live fungal units in eight different isolates . Cfu on BHIA + HS + EXT plates showed an excellent plating efficiency over 96% in all isolates tested . The percentage of the live cells indicated by fluorescent staining (FL) or vital dye exclusion test with erythrosin B (EB) or Janus green (JG-1) was lower than that of cfu . By contrast, the percentage due to modified dye exclusion test with Janus green (JG-2) and that due to lactophenol cotton blue staining (LPCB) showed a close correration to that of cfu . Our results indicate that the modified dye exclusion test with Janus green and lactophenol cotton blue staining are useful for estimating cell viability of yeast form cells of P . brasiliensis.

J Comput Assist Tomogr, 1993 Nov-Dec, 17(6), 909 - 14
CT-guided paracoccygeal drainage of pelvic abscesses; Longo JM et al.; OBJECTIVE: Using the transrectal/transvaginal routes for the drainage of pelvic abscesses complicating colorectal surgery (anterior resection or abdominoperineal resection) is not always possible . The conventional transgluteal approach through the greater sciatic foramen, although proven to be a valuable access route, can have complications (mainly local pain) . MATERIALS AND METHODS: To avoid these difficulties, a CT-guided paracoccygeal-infragluteal approach was used in the percutaneous drainage of deep pelvic (presacral and ischiorectal) abscesses presenting after colorectal surgery in six patients . RESULTS: Percutaneous drainage through this approach was successful in preventing the need for surgery in all six patients . No complications or recurrences were noted, and catheters were removed an average of 15 days after insertion . CONCLUSION: In comparison with the classical transgluteal approach, the paracoccygeal-infragluteal approach minimizes patient discomfort and minimizes the risk of potential injury to the sciatic plexus or blood vessels . This initial series shows that a CT-guided paracoccygeal-infragluteal approach is well tolerated, safe, and effective for the percutaneous drainage of pelvic abscesses developing after colorectal surgery.

Rev Inst Med Trop Sao Paulo, 1993 Nov-Dec, 35(6), 535 - 41
{Comparative study of the pathogenicity and antigenicity of 6 Paracoccidioides brasiliensis strains}; Finquelievich JL et al.; A comparative study of antigenicity and pathogenicity for rats of six Paracoccidioides brasiliensis strains was carried out . The antigenic capacity "in vitro" of cytoplasmic extract from each strain was determined by immunodiffusion test against 6 serum samples obtained from rats experimentally infected with Paracoccidioides brasiliensis, that had presented positive reactions with a metabolic control antigen . The cytoplasmic extracts were used at final concentration of 100 mg/ml . All of them showed 2 or 3 precipitation bands in this assay . One hundred twenty Wistar rats both sexes weighing approximately 200 g, were inoculated intracardiacally with suspensions of the yeast phase of different P . brasiliensis strains . Two concentrations containing 3 x 10(7) and 5 x 10(7) cells/ml of each isolate were prepared . The inoculated animals were divided in two groups, one was left to its spontaneous outcome and the percentages of deaths were registered and the other rats were sacrificed at 14, 28, 56 and 70 days post-infection . The following parameters were taken into account for evaluation: A) presence of macroscopic granulomas in lung, liver, spleen and kidney; B) presence of P . brasiliensis in microscopic exams of the same organs, in wet preparations and in histologic sections stained by H&E; C) culture of lung and D) immunodiffusion test using pre-mortem serum samples and the homologous antigen . The correlation between the most important parameters studied in each strain are summarized as follow: As no significant differences between the two inocula employed for each strain was observed, the before mention results are the average of those obtained with each inoculation doses.(ABSTRACT TRUNCATED AT 250 WORDS)

Clin Exp Immunol, 1993 Oct, 94(1), 75 - 9
Experimental murine paracoccidioidomycosis: relationship among the dissemination of the infection, humoral and cellular immune responses; Singer-Vermes LM et al.; The dissemination of Paracoccidioides brasiliensis cells to the heart, omentum/pancreas, spleen, liver and lungs, assessed by colony forming unit (CFU) counts, the levels of specific antibodies to this fungal agent (by ELISA), and the specific DTH reaction were studied in susceptible (B10.A) and resistant (A/Sn) mice . The animals were infected intraperitoneally with P . brasiliensis yeast cells and were evaluated 2, 4, 12 and 16 weeks later . The most remarkable differences between the two mouse strains were observed 16 weeks after infection, when B10.A mice displayed high numbers of CFU in all examined organs, except the heart, high antibody titres, and depressed DTH response . At this point, A/Sn mice presented low or absent CFU in all organs, low antibody titres and expressive DTH response . The CFU counts were shown to be a reliable parameter to discriminate susceptible from resistant animals . The fungal load in the most affected organs correlated with the antibody titres and was inversely correlated with the intensity of the DTH reaction . The patterns of immune response in this model mimic human paracoccidioidomycosis, in which high specific antibody levels and depressed DTH reactions are found in multifocal and severe forms of the disease.

Clin Exp Immunol, 1993 Oct, 94(1), 11 - 6
Effect of anti-IL-4, interferon-gamma and an antifungal triazole (SCH 42427) in paracoccidioidomycosis: correlation of IgE levels with outcome; Hostetler JS et al.; Paracoccidioidomycosis is characterized by depressed cellular but enhanced humoral immune responses, which suggests a Th2 type of response to infection . We investigated possible therapeutic roles for anti-IL-4, interferon-gamma (IFN-gamma) and/or SCH 42427 (SCH), a new triazole antifungal agent, and their effect on serum IgE levels in a murine model of chronic Paracoccidioides brasiliensis infection . BALB/c mice infected by the pulmonary route were studied with three programmes . The subacute model and one acute model experiment investigated cytokine secretion by lymph node cells (LNC), and in a second acute experiment mice were given anti-IL-4, IFN-gamma or nothing 24 h post infection, then killed at 4 weeks . In the chronic model, mice began treatment at 4 weeks post infection, receiving either SCH, IFN-gamma alone, SCH+IFN-gamma, or no treatment for 8 weeks . At 2-week intervals lung and spleen burdens of infection and serum polyclonal IgE levels were determined . In the subacute model (non-progressive infection), initially there was dual production of IL-4 and IFN-gamma by antigen-stimulated LNC . In the acute progressive infection model IL-4, but not IFN-gamma, was secreted . Anti-IL-4 treatment of the acute phase resulted in enhanced host resistance to infection, which correlated with decreased serum IgE . The chronic model, in which the in vivo efficacy of SCH against P . brasiliensis was shown, suggests possible synergy between immunomodulation and antimicrobial chemotherapy (IFN-gamma and SCH) . Decreased organ burdens of infection in the chronic model after treatment with SCH, SCH plus IFN-gamma, or anti-IL-4 correlated with decreased serum IgE . These promising novel approaches to treatment of systemic fungal infections suggest a Th2 type of response to P . brasiliensis infection, which can be reversed with successful therapy.

Mycopathologia, 1993 Oct, 124(1), 7 - 11
Immunohistochemical characterization of mononuclear cells in delayed hypersensitivity reactions to Paracoccidiodes brasiliensis (paracoccidioidin test); Marques M et al.; The density and distribution of T cells, T helper cells, macrophages and B cells at the site of skin tests with a cytoplasmic Paracoccidioides brasiliensis antigen (paracoccidioidin) was studied at 24 and 48 h post-challenge in 10 patients with the chronic form of paracoccidioidomycosis and in 5 non-infected individuals . The in situ study was carried out using immunoperoxidase techniques and monoclonal antibodies . The controls showed negative skin test . In the patients, the great majority of the cells in the perivascular foci were T cells (CD43-positive cells) making up 47% and 48.6% of the total number of cells at 24 and 48 h respectively . Most of the T cells showed a T helper phenotype (CD45RO-positive cells) . Approximately 25% of the cells were macrophages (CD68-positive cells) and there were very few B lymphocytes (CD20-positive cells) . The present data on the microanatomy of paracoccidioidin skin test sites were consistent with a delayed type hypersensitivity pattern . Our results were comparable to those reported on skin tests for other granulomatous chronic diseases.

Mycopathologia, 1993 Sep, 123(3), 131 - 3
Agarose gel immunodiffusion tests with the addition of PEG (polyethylene glycol 6000) for the diagnosis and serological follow up of paracoccidioidomycosis infection; Forjaz MH et al.; By using PEG (polyethylen glycol 6000) in the gel immunodiffusion tests (ID), the precipitin lines were increased in 25.5% of the 192 sera reactions and the titers were increased from one to four serial dilutions in 44.6% of the 139 serum samples . Owing to its sensitivity, easy interpretation of the results and low cost, the use of 2% PEG incorporated into the gel in ID tests is recommended for the diagnosis and serological follow-up of paracoccidioidomycosis infections.

Mycoses, 1993 Sep-Oct, 36(9-10), 283 - 7
Specific digestive deficiency of phagocytes in paracoccidioidomycosis . Its absence in peripheral blood neutrophils of members of the nuclear family of patients . An initial report; Urquiola G et al.; Paracoccidioidomycosis (PARA) affects only a minority of individuals, who have presumably been exposed to the causative fungus (Paracoccidioides brasiliensis) . Neutrophils (PMNs) from patients with PARA show a significant and specific digestive deficiency phagocytosed P . brasiliensis in vitro . It is not known whether the defect is acquired after contact with the fungus, or precedes it . We studied the spouses and offspring of three patients with PARA . Individuals studied stayed in the same house as their husband or father . None of the relatives had evidence of PARA, and their PMNs showed no defect in their ability to digest or kill the fungus . Relatives showed no indication of sensitization against P . brasiliensis . These results are compatible with the view that effective contact with P . brasiliensis may occur only under restricted conditions and that the defect in digestive ability is an infrequent and specific occurrence.

Epidemiol Infect, 1993 Aug, 111(1), 121 - 33
Factors associated with Paracoccidiodes brasiliensis infection among permanent residents of three endemic areas in Colombia; Cadavid D et al.; The natural habitat of Paracoccidioides brasiliensis, the aetiologic agent of paracoccidioidomycosis, has not been determined . Consequently, the events leading to the acquisition of infection remain controversial . To identify factors associated with infection in endemic areas we conducted a survey in three rural communities in Colombia where we had previously diagnosed paracoccidioidomycosis in children . Permanent residents were surveyed taking into consideration environmental and occupational variables . Skin tests were used to classify subjects as infected or non-infected . Variables found associated with infection were: (i) community A: previous residence around Porce river and agriculture in vegetable gardens; (ii) community C: frequent use of specific water sources; (iii) community V: housekeeping activities, and (iv) total group: age > 25 years and contact with bats . Residents in communities with higher prevalence of infection were older, had more complex residence history, and referred more contact with armadillos than residents of communities with lower infection.

Clin Exp Immunol, 1993 Aug, 93(2), 189 - 94
Tumour necrosis factor production in vivo and in vitro in response to Paracoccidioides brasiliensis and the cell wall fractions thereof; Figueiredo F et al.; Tumour necrosis factor (TNF) was detected in serum from mice challenged with Paracoccidioides brasiliensis . The serum TNF levels of mice challenged with an avirulent strain were significantly higher than those of mice challenged with a virulent strain, and the same was observed for the TNF levels of mice challenged with a cell wall fraction (F1) from the two fungal strains . Fraction F1 consisted of chitin and beta-glucan; but although the chitin contents were similar for the two strains, the avirulent strain allowed a greater content of beta-glucan . The beta-glucan, purified from both strains, increased serum TNF levels in an identical dose-dependent manner, whereas purified chitin did not induce serum TNF levels . P . brasiliensis, the F1 fractions and beta-glucan induced macrophages to secrete TNF in vitro . The differences in TNF levels, induced by the different fungal strains, were correlated with the beta-glucan concentrations in the cell walls of both the avirulent and virulent strains of P . brasiliensis . These findings support a role for TNF in the pathogenicity of P . brasiliensis.

Rev Soc Bras Med Trop, 1993 Jul-Sep, 26(3), 141 - 3
{The association between glyoxalase I and paracoccidioidomycosis infection}; Balarin MA et al.; With the purpose to study the genetic susceptibility to paracoccidioidomycosis infection we searched for a possible association between glyoxalase I and the intradermic paracoccidioidin reaction . The phenotype GLO 1 was significantly more frequent among positive reactors.

J Nucl Med, 1993 Jun, 34(6), 958 - 60
Successful gallium-67 imaging of North American pulmonary blastomycosis; Dayanikli BF et al.; South American blastomycosis (Paracoccidioidomycosis) has been previously imaged using 67Ga imaging . We present a case of North American pulmonary blastomycosis successfully imaged with 67Ga . Clinical, radiologic and biopsy findings are correlated.

Am J Trop Med Hyg, 1993 May, 48(5), 626 - 9
Recovery of adrenal function after treatment of paracoccidioidomycosis; Do Valle AC et al.; In endemic areas, the systemic mycosis paracoccidioidomycosis (PCM) is the most frequent etiology of Addison's disease . Paracoccidioides brasiliensis, the causative agent of PCM, exhibits a high tropism for the adrenal glands, which results in a low hormone reserve and in more severe cases, in symptoms of primary adrenal insufficiency . In these cases, the hormone deficit is usually treated with replacement corticoid therapy for the rest of the patient's life . Recently, we identified three patients with disseminated PCM who had adrenal insufficiency; one of them had Addison's disease . All showed complete recovery of adrenal function after a 1-2-year period of specific therapy with ketoconazole or sulfonamides . Plasma cortisol levels were determined by radioimmunoassay of baseline blood samples, as well as those taken after the rapid stimulation test with synthetic adrenocorticotrophic hormone, and before and after the antifungal therapy.

Clin Microbiol Rev, 1993 Apr, 6(2), 89 - 117
Paracoccidioidomycosis: an update; Brummer E et al.; This review summarizes knowledge on various aspects of paracoccidioidomycosis . Mycelial propagules, chlamydospores, and arthroconidia exhibit thermal dimorphism; arthroconidia are infectious in animals and, by electron microscopy, appear well provided for survival . The mycelial-to-yeast-phase transformation requires a strict control of glucan synthesis probably mediated by membrane enzymes . Hormonal influences on the transformation of the fungus (mycelium or conidium to yeast phase) have been demonstrated . Estrogen-binding proteins have been detected in the fungal cytosol, and during the transformation novel proteins are produced as a result of estradiol incorporation . Clinical forms have been better defined on the basis of better experimental models . Emphasis has been placed on the lungs as the portal of entry and on the existence of silent pulmonary infections . A specific Paracoccidioides brasiliensis antigen, the 43-kDa glycoprotein (Gp43), has been identified, characterized, and cloned . This has led to improved reproducibility and specificity of serologic tests . The depression of cell-mediated immune responses has been associated with severe disease in humans and in the experimental host . T-cell subsets in patients' tissues were characterized by means of monoclonal antibodies, and a reduced CD4/CD8 ratio was demonstrated . This has been related to alterations in lymphokine and tumor necrosis factor production, production of antigen-antibody complexes, etc . Amphotericin B has provided effective therapy . Azole derivatives have also improved prognosis and facilitated therapy . Itraconazole is presently the drug of choice, yet incapacitating sequelae (mainly pulmonary fibrosis) still constitute major problems.

Oral Surg Oral Med Oral Pathol, 1993 Apr, 75(4), 461 - 5
Oral paracoccidioidomycosis . A study of 36 South American patients; Sposto MR et al.; Paracoccidioidomycosis (South American blastomycosis) is an uncommon, progressive systemic mycosis, virtually only seen in persons who have visited Latin America . Reports of oral lesions are extremely rare in the English-language literature . Thirty-six adults with oral lesions as the first sign of paracoccidioidomycosis are described; this appears to be the largest series in the dental literature . All had chronic proliferative mulberry-like ulcerated oral lesions; the diagnosis was confirmed histologically . The gingiva or alveolar process was the typical site, but lesions were also seen particularly on the palate and lip . Most of the patients proved to have detectable pulmonary involvement . Patients with lesions in the oropharynx, tongue, or floor of mouth all had confirmed pulmonary lesions.

Mycopathologia, 1993 Apr, 122(1), 7 - 13
Two-dimensional immunoelectrophoresis of histoplasmin and a purified polysaccharide-protein antigen of Histoplasma capsulatum; Toriello C et al.; Crude histoplasmin and a polysaccharide-protein complex (PPC-histo) antigens obtained from culture filtrates of Histoplasma capsulatum were analyzed by single and tandem two-dimensional immunoelectrophoresis (TD-IEP) using a rabbit hyperimmune anti-histoplasmin polyvalent serum . Single TD-IEP showed 14 arc precipitates for histoplasmin . Continuity of arcs 2, 6, and 7, and 9 and 10 was observed, suggesting a different polymeric configuration of the same antigen . This was also confirmed in tandem TD-IEP of histoplasmin with homologous (PPC-histo) and heterologous PPC's from Blastomyces dermatitidis, Paracoccidioides brasiliensis and Coccidioides immitis . Tandem TD-IEP of histoplasmin and PPC-histo displayed a similar antigenic pattern to histoplasmin alone, being arcs 1 and 3 more evident and apparently present only in histoplasmin and PPC-histo . Tandem TD-IEP showed common antigens among the other heterologous fungal purified antigens, and seems useful to observe the multiplicity of antigens present in fungal preparations and to identify those precipitates (arcs 1 and 3) that are predominant in the purified preparation.

FEMS Microbiol Lett, 1993 Mar 15, 108(1), 31 - 4
Identification of sialic acids on the cell surface of hyphae and yeast forms of the human pathogen Paracoccidioides brasiliensis; Soares RM et al.; Paracoccidioides brasiliensis, the agent of paracoccidioidomycosis, when grown in a synthetic medium, expresses at the cell surface of both yeast and mycelial forms acidic glycoconjugates containing N-acetylneuraminic acid units . Sialic acids were extracted using mild hydrolytic conditions, and were identified by thin-layer and gas chromatography, standard colorimetry, reaction with periodate-resorcinol and mass spectrometry . Their surface location was inferred from fluorescent-lectin (Limulus polyphemus agglutinin) binding to whole cells abrogated by previous treatment with neuraminidase . Expression of sialic acids on virulent yeast forms of P . brasiliensis (3.7 x 10(6) residues per cell) may inhibit fungal phagocytosis during early infection, when the immunological response is still being built up.

Mycopathologia, 1993 Mar, 121(3), 149 - 56
Dialysable leukocyte extracts modify the course of experimental paracoccidioidomycosis in the Syrian hamster; Peracoli MT et al.; The effect of dialysable leukocyte extracts (DLE) obtained from hamsters immunized with Paracoccidioides brasiliensis (immune DLE) and from non-immunized hamsters (non-immune DLE) was studied in hamsters inoculated with P . brasiliensis by the intratesticular route . Treatment with immune or non-immune DLE was started during the third week of infection and was repeated at 7, 11, 15 and 19 weeks . A group of untreated infected animals was used as control . Animals were submitted to the delayed hypersensitivity skin test to P . brasiliensis antigen (PbAg) in vivo and assayed in vitro by the macrophage migration inhibition test in the presence of Phytohemagglutinin (PHA) and PbAg and by immunodiffusion for specific antibody . The animals were sacrificed at 4, 8, 12, 16 and 20 weeks . The morphology and extension of the lesions were studied at the inoculation site, and in lymph nodes, lungs, liver, spleen and kidneys . In contrast to the controls, animals treated with both DLEs maintained a positive cell-mediated immune response throughout the experiment and developed less extensive infection with a significantly lower number of fungi in the lesions . The results suggest that immune and non-immune DLE preparations modified the evolution of experimental paracoccidioidomycosis with equal efficiency . This similarity may be explained by the immunoregulatory activities of both extracts.

J Clin Microbiol, 1993 Mar, 31(3), 671 - 6
Immunological response to cell-free antigens of Paracoccidioides brasiliensis: relationship with clinical forms of paracoccidioidomycosis; Blotta MH et al.; Sera from patients with the acute (AF) and chronic (CF) forms of paracoccidioidomycosis (PCM) were tested against Paracoccidioides brasiliensis cell-free antigens by Western blot (immunoblot) . The CFA preparation contained components ranging in molecular mass from 18 to 102 kDa . The immunoglobulin G (IgG) reactivity profiles were similar for patients with both forms of the disease, and the 43-kDa component was recognized by 100% of the sera . IgM antibodies from the AF- and the CF-PCM sera recognized 21 and 20 components, respectively, the AF-PCM sera reacting preferentially with components with molecular masses above 50 kDa . None of the AF-PCM sera (IgM) reacted with the 43-kDa component, and only 10% of the CF-PCM sera recognized this molecule . The IgA response was more significant in the CF-PCM group than in the AF-PCM group, and the 43- and 74-kDa components were the most reactive ones (about 40% each) . Our results showed that the cell-free antigen preparation is very appropriate for the immunoblotting analysis of PCM sera, and they also showed that the detection of IgG anti-gp43 is the best marker for the diagnosis and the following up of patients with the acute or the chronic form of the disease.

Rev Hosp Clin Fac Med Sao Paulo, 1993 Mar-Apr, 48(2), 94 - 7
{Paracoccidioidomycosis of the central nervous system: study of 5 cases by magnetic resonance}; Magalhaes AC et al.; Paracoccidioidomycosis is a systemic mycosis, endemic in South and Central America, that affects the central nervous system in almost 10% of the cases . Neurological involvement includes to different clinical forms: meningeal and granulomatous form, also known as pseudotummoral form . Five patients with intracranial paracoccidioidomycosis were submitted to MR examination in a 1.5T system . All patients presented multiple round or lobulated lesions, hyperintense in T1-weighted images and predominantly hypointese in T2-weighted images . The lesions were distributed diffusely, with a slight predominance in the supratentorial compartment, although infratentorial lesions were also observed, mainly in the cerebellum . All lesions had ring or nodular contrast enhancement . Three patients were reexamined for treatment control, with disappearance of the lesions in two, with persistence of hypointense area in T2-weighted images . MR imaging proves to be a good method to evaluate the neurological involvement in paracoccidioidomycosis, mainly due to its high sensitivity for posterior fossa lesions . Further studies are needed to correlate the MR findings and the changes found in MR examination after treatment.

J Med Vet Mycol, 1993, 31(2), 99 - 113
Paracoccidioidomycosis and its etiologic agent Paracoccidioides brasiliensis; San-Blas G; Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis, a systemic mycosis endemic to Latin America . In this paper we review clinical, therapeutic and immunological aspects of the disease, as well as the biology and ecology of the fungus, restricting the review to the period 1989-1992.

J Med Vet Mycol, 1993, 31(2), 155 - 60
Diagnosis of paracoccidioidomycosis by passive haemagglutination assay of antibody using a purified and specific antigen-gp43; Taborda CP et al.; Sheep red blood cells, treated with glutaraldehyde and sensitized with a purified Paracoccidioides brasiliensis antigen of molecular mass 43 kDa (gp43), were used to detect antibodies to P . brasiliensis by passive haemagglutination (PHA) . Sera from 50 patients with paracoccidioidomycosis had PHA titres of > 1/800, and had antibodies detectable by immunodiffusion while antibodies to gp43 were detectable by PHA in sera from patients with histoplasmosis, aspergillosis, candidiasis, Jorge Lobo's disease or healthy people, only at titres of < 1/100 . Considering a cut-off at 1/200, the sensitivity and specificity of PHA for paracoccidioidomycosis was 100% . Thus, the PHA test could be an efficacious method for routine serological diagnosis of paracoccidioidomycosis, with results in 1 to 2 h.

J Med Vet Mycol, 1993, 31(2), 133 - 41
Effect of ajoene on dimorphism of Paracoccidioides brasiliensis; San-Blas G et al.; Ajoene, a compound derived from ethanolic garlic extracts, inhibits the growth of the dimorphic pathogenic fungus Paracoccidioides brasiliensis, yeast cells being more sensitive to its action than mycelial cultures . Sulphydryl protector groups were able to cancel the inhibitory effect on the yeast cells but not on the mycelial cultures . The dimorphic process was also blocked when mycelia were placed to transform to yeast . Synthesis in vitro of cell wall glucan was not affected by ajoene . We discuss the possible involvement of ajoene on sulphydryl metabolism of P . brasiliensis.

J Med Vet Mycol, 1993, 31(1), 17 - 27
The role of the complement system in the neutrophil functions stimulated in vitro by an alkali-insoluble cell wall fraction of Paracoccidioides brasiliensis; Crott LS et al.; We investigated the capacity of an alkali-insoluble cell wall polysaccharide fraction (F1) of Paracoccidioides brasiliensis to induce rat polymorphonuclear neutrophil (PMN) migratory and chemiluminescence (CL) responses . Normal rat serum pre-incubated with F1 induced a chemotactic neutrophil response which was fully abolished by heat-inactivation . The participation of the alternative complement pathway was more effective than that of the classical pathway since depletion of factor B by heating at 50 degrees C reduced PMN migration, whereas blockade of the classical pathway with EGTA left the migratory response practically unchanged . Opsonized serum F1 induced a significant release of oxygen radicals from PMN as measured by CL . The complement system was also found to be involved in this activity since serum inactivation at 56 degrees C altered the CL response . In addition to complement-derived fragments, other serum opsonins, probably cross-reacting antibodies, were required for optimal interaction between PMN and opsonized particles . These results contribute to the understanding of the role of fungal components and of the complement system in the inflammatory response observed in paracoccidioidomycosis.

Mycopathologia, 1993 Jan, 121(1), 1 - 5
Cell wall fractions from Paracoccidioides brasiliensis induce hypergammaglobulinemia; de Oliveira SL et al.; The antibody response against the antigen sheep red blood cells (SRBC) was investigated in mice pre-treated with formalin-killed Paracoccidioides brasiliensis or with cell wall fractions of the fungus . Pre-treatment with P . brasiliensis, as well as with the Fl fraction and beta-glucan significantly increased the anti-SRBC antibody response in the experimental groups as compared to the control group that received only SRBC . This immunomodulatory effect varied with the different doses employed and with pre-treatment time . We conclude that the cell wall fractions of P . brasiliensis might play an important role in the hypergammaglobulinemia associated with Paracoccidioidomycosis.

J Med Vet Mycol, 1993, 31(2), 169 - 74
A preliminary study of in vitro antibiotic activity of saperconazole and other azoles on Paracoccidioides brasiliensis; San-Blas G et al.; Saperconazole, a new triazole related to itraconazole, was tested against Paracoccidioides brasiliensis and results compared with ketoconazole and itraconazole . The fungus was highly sensitive to the action of these compounds, particularly saperconazole, with minimum inhibitory concentrations and minimum fungicidal concentrations ranging from 10(-7) to 10(-10) M (equivalent to 6.7 x 10(-2)-6.7 x 10(-5) micrograms ml-1 for saperconazole), according to the morphological phase and the antifungal tested . The yeast phase was more sensitive than the mycelial phase to any of the azoles . Morphological changes were observed in the cell membranes, particularly when saperconazole was used as the antifungal agent.

J Med Vet Mycol, 1993, 31(3), 249 - 57
Placental involvement in paracoccidioidomycosis; Blotta MH et al.; A case of juvenile-type paracoccidioidomycosis in a pregnant woman is reported . The disease pre-dated pregnancy and antifungal treatment was being administered when she became pregnant . A premature male infant was delivered with no evidence of infection . Microscopic examination of the placenta showed numerous Paracoccidioides brasiliensis yeast forms in the intervillous space, enmeshed in a macrophagic-phagocytic reaction, with damage of the trophoblastic layer . Placental transfer of specific P . brasiliensis antibodies was demonstrated.

J Med Vet Mycol, 1993, 31(3), 201 - 5
An improved culture medium for detecting live yeast phase cells of Paracoccidioides brasiliensis; Kurita N et al.; The plating efficiency of standard mycological media such as brain heart infusion (BHI) agar is poor for Paracoccidioides brasiliensis . We prepared a water-extract of yeast phase cells of P . brasiliensis and examined it for growth-enhancing activity for the fungus . The water-extract, when added to BHI agar to a concentration of 5%, improved the plating efficiency of the medium for the fungus to some extent, but the degree of improvement was considerably varied among P . brasiliensis isolates . By contrast, when the water-extract was added in combination with horse serum (4%), the plating efficiency was highly improved (to 94-99%) for all the P . brasiliensis isolates employed . The growth-enhancing factor(s) in the water-extract was heat-stable and heating at 120 degrees C for 15 min had little, if any, effect on growth-enhancing activity.

Arch Med Res, 1993 Autumn, 24(3), 267 - 8
Biochemical and physiological aspects in the dimorphism of Paracoccidioides brasiliensis; San-Blas G; Paracoccidioides brasiliensis is a dimorphic fungus pathogenic for humans . It shows a yeast like phase at 37 degrees C and a mycelial phase at 23 degrees C . Biochemical aspects of its dimorphism are related to the presence of hormone receptors, regulation of cAMP, or modulation of glucan synthetase activity through cytoplasmic proteinases . These aspects are reviewed herein.

Arch Med Res, 1993 Autumn, 24(3), 239 - 45
Advances in experimental paracoccidioidomycosis using an isogenic murine model; Singer-Vermes LM et al.; A genetically controlled murine model of paracoccidioidomycosis which allowed us to investigate several parameters of the host-parasite interactions was established in our laboratory . Natural resistance and acquired immune responses to P . brasiliensis infection were investigated employing resistant and susceptible mice infected with highly virulent or slightly virulent P . brasiliensis isolates . Resistant mice inoculated with a highly virulent P . brasiliensis isolate present efficient macrophage activation, presence of DTH response, low levels of specific antibody and a tendency to resolution of the infectious process, suggesting that a T helper-1 mode of immune response is mounted . Susceptible mice, on the contrary, seem to mount a predominantly T helper-2 type of immune response activation in which an inefficient macrophage activation, depressed DTH reactions and high levels of antibodies result in progressive disease . The crucial role of the fungal virulence on the outcome of the infection of susceptible and resistant mice is also demonstrated, thus reinforcing the idea that both the innate resistance of the host and the pathogenicity of the fungal cells are determinant on the outcome of the disease . This model is proposed as a framework of our current knowledge of the host-parasite interactions in paracoccidioidomycosis and as a basis for future challenge in continuing analyses.

Mem Inst Oswaldo Cruz, 1993 Jan-Mar, 88(1), 111 - 7
Histopathological alterations induced by non-viable cells and biochemical fractions from Paracoccidioides brasiliensis in mice; Hamdan JS et al.; Non-viable cells and biochemical fractions from Paracoccidioides brasiliensis were obtained for experimental inoculation in mice and posterior histopathological analysis . Dead total fungus, total fungus disrupted by sonorous waves, lipids of the fungus, supernatant of the lipid purification, integral and disrupted fungus free of lipids were obtained . The six preparations arose from masses of lyophilized yeasts of a recent isolate of P . brasiliensis (strain JT-1) and from a "Pool" equitably constituted by four strains maintained in laboratory for a long time (SN, 2, 18 and 192) . Different doses of the 12 preparations were intraperitoneally inoculated and histopathological analysis were done 30 days later . This analysis showed that all the inoculated preparations gave origin to inflammatory foci, except the one designated "supernatant of lipid purification" . The alterations were detected exclusively in the liver of the animals and occurred from the smallest dose tested (1 mg), with exception of the lipids of the fungus, where the foci appeared only from a 3 mg dose onwards . No difference in the capacity of inducing histopathological alterations was found between the preparations obtained from the recent isolate (JT-1) and from the older ones ("Pool") . On the other hand, an increase of the number of inflammatory foci in function of the inoculated dose was observed.

J Clin Microbiol, 1992 Dec, 30(12), 3108 - 11
Comparative evaluation of a chemiluminescent DNA probe and an exoantigen test for rapid identification of Histoplasma capsulatum; Padhye AA et al.; A chemiluminescent DNA probe (Accuprobe) assay developed by Gen Probe, Inc., for the rapid identification of Histoplasma capsulatum was evaluated and compared with the exoantigen test by using 162 coded cultures including Histoplasma capsulatum var . capsulatum, Histoplasma capsulatum var . duboisii, Histoplasma capsulatum var . farciminosum, Blastomyces dermatitidis, Coccidioides immitis, Paracoccidioides brasiliensis, and morphologically related saprobic fungi . Each test uses a chemiluminescent, acridinium ester-labeled, single-stranded DNA probe that is complementary to the rRNA of the target organism . Lysates of the test cultures were prepared by sonication with glass beads and heat treated . After the rRNA was released from the target organism, the labeled DNA probe combined with the target H . capsulatum rRNA to form a stable DNA-RNA hybrid . A hybridization protection assay was used, and the chemiluminescence of hybrids was measured initially with a Leader 1 luminometer as relative light units and later during the investigation with a probe assay luminometer as probe light units . Of the 162 coded mycelial cultures tested by the Accuprobe assay, 105 were identified as H . capsulatum . The test could be performed with an inoculum of a few square millimeters (1 to 2 mm2) of growth . In the primary evaluation, the Accuprobe identified 103 of the 105 cultures as H . capsulatum within 2 h . The remaining two cultures, contaminated with bacteria, had to be purified before the Accuprobe assay identified them correctly as H . capsulatum . Since each coded culture was concurrently tested for H . capsulatum, B . dermatitidis, and C . immitis exoantigens, the identification of all three dimorphic pathogens was provided simultaneously . Of the 162 coded cultures tested, 105 were identified by the exoantigen test as H . capsulatum, 12 were identified as B . dermatitidis, 13 were identified as C . immitis, and 32 were negative for H . capsulatum, B . dermatitidis, and C . immitis . The bacterial contamination in two isolates did not interfere with the exoantigen testing . The exoantigen test required 7- to 10-day-old colonies and required 48 to 72 h of incubation before definitive identification was obtained.

Rev Inst Med Trop Sao Paulo, 1992 Nov-Dec, 34(6), 565 - 7
{Paracoccidioides brasiliensis (Lutz, 1908) isolated by hemoculture in a patient with the acquired immunodeficiency syndrome (AIDS)}; Hadad DJ et al.; A case of an AIDS patient with positive blood culture for Paracoccidiodes brasiliensis is reported . The patient was a 29-years-old male born in Nova Londrina (Parana State, Brazil) who presented splenomegaly and fever of unknown origin . Three blood cultures were performed, each one of them for aerobic bacteria, mycobacteria and fungi . Cultures for aerobic bacteria and mycobacteria were negative . However, the yeast phase P . brasiliensis was isolated from two cultures in BHI agar, 20 days after inoculation in Negroni medium . The patient was classified in group V according to the Centers for Disease Control (CDC) criteria for AIDS, due to a Pneumocystis carinii pneumonia . Treatment was discontinued due to an individual decision of the patient on the second dose of amphotericin B . This antibiotic was replaced by ketoconazole in the daily dose of 800 mg . The patient died one year after the isolation of P . brasiliensis on blood culture.

Rev Inst Med Trop Sao Paulo, 1992 Nov-Dec, 34(6), 511 - 6
Biochemical analysis of the methylic antigen of Paracoccidioides brasiliensis; Hamdan JS et al.; Yeast forms of five strains of Paracoccidioides brasiliensis (SN, 2, 18, 192 and JT-1) were cultured in a synthetic medium for obtaining methylic antigens . These antigens were lyophilized and studied for each strain, to determine their partial biochemical composition, through measurements of total lipid, protein and carbohydrate contents . Lipids of methylic antigens were purified and analysed for sterols, phospholipids, glycolipids, lipoproteins, and partial characterization of sterols . Significant differences were found among antigenic preparations derived from distinct P . brasiliensis strains, in relation to the quantitative determinations . On the other hand, sterol analysis revealed the presence of ergosterol, lanosterol and squalene in all samples . The diversity verified in the biochemical characteristics of antigens derived from different P . brasiliensis strains, confirm the need of using a pool of fungal samples in order to produce antigen preparations for serological procedures without hampering their sensitivity.

Mycoses, 1992 Nov-Dec, 35(11-12), 275 - 80
Haematological alterations induced by biochemical fractions of Paracoccidioides brasiliensis in mice; Hamdan JS et al.; Biochemical fractions of Paracoccidioides brasiliensis were obtained for experimental inoculation in mice followed by haematological analysis . Dead total fungus, total fungus disrupted by ultrasonic waves, lipids of the fungus, supernatant of the lipid purification, and integral and disrupted fungus free of lipids were obtained . The six fractions were obtained from lyophilized yeasts of a recent isolate of P . brasiliensis and from a pool comprising equal amounts of four strains maintained in the laboratory for some time . Different doses of the 12 fractions were intraperitoneally inoculated into mice and haematological analysis was done 30 days later . No significant alterations were detected in the red blood cell series . However, the white blood cell series showed marked alterations, such as leukopenia, with relative neutrophilia and lymphopenia . Thrombocytosis occurred widely . The haematological alterations revealed associations with the injected doses without relation to the biochemical composition of the different fractions . No difference in the capacity for inducing haematological alterations was found between the fractions obtained from the recent isolate and from the older ones.

Mycoses, 1992 Nov-Dec, 35(11-12), 269 - 74
Relationship between digestive and killing abilities of neutrophils against Paracoccidioides brasiliensis; Goihman-Yahr M et al.; Peripheral blood neutrophils (PMN) from patients with paracoccidioidomycosis killed and digested Paracoccidioides brasiliensis much less than did PMN from normal individuals or from patients with other diseases . However, deficiency in killing ability was less specific than digestive deficiency and correlated poorly with it . We conclude that the capacities of PMN to digest and kill P . brasiliensis are not intimately related phenomena, and that in paracoccidioidomycosis the key deficiency of neutrophil function is that of digestion of P . brasiliensis.

Int J Dermatol, 1992 Oct, 31(10), 725 - 9
Saperconazole in the treatment of systemic and subcutaneous mycoses; Franco L et al.; In a 2-year period, 30 patients with culture-proven mycoses (chromoblastomycosis, sporotrichosis, and paracoccidioidomycosis) were treated with the new orally administered triazole, saperconazole (SPZ) (R66905) . The daily dose varied from 100 to 200 mg . All patients responded to treatment; the mean time required to heal the lesions and convert the cultures to negative was 3.5 months for sporotrichosis, 4.6 for paracoccidioidomycosis, and 9.0 for chromoblastomycosis . Evaluation by a scoring system indicated that 36.6% of the patients achieved complete resolution of the pretherapy abnormalities, while the remaining (63.3%) experienced major improvement . No collateral effects were reported; there were no bone-marrow or liver toxicities . SPZ is an effective drug for the treatment of the above-mentioned mycoses and appears to be suitable for the control of chromoblastomycosis.

Arq Gastroenterol, 1992 Oct-Dec, 29(4), 147 - 52
{Gastric paracoccidioidomycosis . A case report and considerations on the pathogenesis of this disease}; Gabellini Gde C et al.; The authors report an unusual case of fungal gastric lesion in a patient with the chronic form of paracoccidioidomycosis . For a period of 8 months the major symptoms of the patient were abdominal pain and body weight loss . The endoscopic evaluation of the upper gastrointestinal tract showed a partial gastrectomy made previously, a great and irregular ulcer in the stomach and a granulated aspect of the duodenal mucosa . A granulomatous process and Paracoccidioides brasiliensis were observed in the histopathological examination of the gastric and duodenal biopsy tissue . New endoscopic evaluation 4 months after treatment with sulfadiazine revealed healing of the gastric ulcer . It was suggested that abdominal lymph nodes involvement, duodenal infection and anatomical and atrophic alterations of the stomach were predisposing factors for the gastric paracoccidioidomycosis.

Mycopathologia, 1992 Sep, 119(3), 141 - 5
Studies on the relationship between the estrous cycle of BALB/c mice and their resistance to Paracoccidioides brasiliensis infection; Sano A et al.; A relationship between the estrous cycle and non-specific host resistance to Paracoccidiodes brasiliensis yeast cells was examined by using both sexes of adult BALB/c mice . They were divided into 6 groups, including a male group and females at proestrus, estrus, metestrus-I, metestrus-II and diestrus . The mice received yeast cells through three different inoculation routes; intravenous, intraperitoneal and intratracheal . In all of the inoculation routes, the clearance of the yeast cells was influenced by the estrous cycle . The female mice at estrus, which might have high blood estrogen levels, showed a marked clearance of the yeast cells from the blood, peritoneal cavity and lungs . These results suggested that non-specific host resistance to the yeast cells was enhanced by estrogen . All female groups inoculated by the three routes showed higher clearance of the yeast cells than the male group.

Mycopathologia, 1992 Aug, 119(2), 65 - 71
Immunosuppressive effect of paracoccidioidomycosis sera on the proliferative response of normal mononuclear cells . Identification of a Paracoccidioides brasiliensis 34-kDa polypeptide in circulating immune complexes; Chequer-Bou-Habib D et al.; In this paper we relate that sera from paracoccidioidomycosis patients inhibited the mitogen-induced proliferative responses of normal mononuclear cells . Treatment of these sera with 2.5% polyethyleneglycol (PEG), a method classically used to precipitate immune complexes, significantly reduced their inhibitory activity . Immunoblot analysis of the PEG precipitates identified a 34-kDa polypeptide, recognized by rabbit anti-P . brasiliensis IgG . Patient mononuclear cells showed partial restoration of their proliferative capacity after 24 h culture in medium alone, which suggests release of membrane-bound molecules in the culture medium . These findings indicate that circulating P . brasiliensis antigens, complexed or not with antibodies, may play a negative immunoregulatory effect in the mitogen-induced proliferative responses of paracoccidioidomycosis patients.

Infect Immun, 1992 Jul, 60(7), 2667 - 71
Isolation and antigenicity of a 45-kilodalton Paracoccidioides brasiliensis immunodominant antigen; Ferreira-da-Cruz MF et al.; In the present study, we analyzed human antibody responses to Paracoccidioides brasiliensis cellular antigens by the immunoblot technique to identify specific cellular components and to investigate the existence of antigen profile differences among serological responses of paracoccidioidomycosis (PCM) patients . Among the 64 PCM serum samples analyzed, a relatively homogeneous immunoglobulin G response to P . brasiliensis antigens was observed . The polypeptide with a mass of 45 kDa was the most clinically important, since antibody to this antigen was detectable in 90.6% of PCM patients studied and the six individuals who did not produce antibody were either at the end of treatment or in the posttherapy period and had shown clinical recovery . These facts suggested that the presence of this antibody may be an indicator of active disease . The 45-kDa antigen was also the most specific antigen of the PCM humoral immune response, since it reacted with only 2 of 79 (2.5%) heterologous serum samples tested: 1 histoplasmosis case and 1 tuberculosis case . This polypeptide was isolated from gels by electroelution and, when tested by an immunoradiometric assay and immunoblotting, maintained its reactivity with PCM sera and also with anti-P . brasiliensis polyclonal antibodies raised in rabbits at the same sensitivity levels as those obtained in immunoblotting with a crude antigen . Since in our assays the 45-kDa polypeptide was the major P . brasiliensis antigen and seemed to be specific for PCM, its use in alternative diagnostic methods is promising, especially in patients suspected of having the juvenile clinical form of PCM often associated with negative double-immunodiffusion results.

Allergol Immunopathol (Madr), 1992 Jul-Aug, 20(4), 145 - 51
Cellular immunity in paracoccidioidomycosis; Oliveira ZN et al.; Paracoccidioidomycosis was studied in 62 patients from Brazil in the 10 year period between 1978 and 1988 . In 46 patients included in a first group, the disease was active and in 16 patients included in a second group, the disease was cured . The study was conducted according to both the clinical form of the disease and the response to paracoccidioidin in both groups . In the first group, 10 patients presented the acute form, 12 presented the chronic unifocal form and 24 had the chronic multifocal form of the disease . As to the response to paracoccidioidin, in the first group 16 patients were negative and 30 were positive; in the second group, 11 were positive and 5 were negative . An immunological study was performed in all patients using in vivo methods such as skin tests and sensitization to DNCB and in vitro techniques such as total lymphocyte counts, T and B cell counts, leukocyte migration inhibition test, chemotaxis of total leukocytes and mononuclear leukocyte phagocytosis.

Mycopathologia, 1992 Jul, 119(1), 1 - 9
Pulmonary paracoccidioidomycosis in immunized mice; Defaveri J et al.; Paracoccidioidomycosis was induced in immunized (IM) and non-immunized (NI) mice . The histopathology, the number of fungi in the lungs, the cellular (footpad test--FPT and macrophage inhibition factor assay--MIF) and humoral (immunodiffusion test) immune response were investigated serially post-infection . In the IM mice, at days 1 and 3, there was intense and predominant macrophagic-lymphocytic alveolitis with loose granulomatous reaction; at day 30, inflammation was mild . In the NI group, up to day 3, the lesions were focal; later there was formation of extensive epithelioid granuloma . The number of fungi in IM mice were always smaller than those of NI group . Immunization alone induced positive FPT and MIF indices with low titer of antibody . After infection, there was a significant decrease of the FPT indices in the IM group, which we interpreted as desensitization due to trapping of sensitized lymphocytes in the lungs . In conclusion, (1) The lesional pattern of pulmonary paracoccidioidomycosis in IM mice was similar to that of a hypersensitivity pneumonitis . This reaction was probably effective in reducing the extension of the infection and decrease the number of fungi . (2) In this model, pulmonary resistance against P . brasiliensis seems to be related to local and systemic delayed-type hypersensitivity reaction.

Rev Soc Bras Med Trop, 1992 Jul-Sep, 25(3), 195 - 200
{Pulmonary lymph node in acute juvenile paracoccidioidomycosis (a case report)}; de Campos EP et al.; The primary complex like Ghon was observed in a child's clinical roentgenographic study . C.S., white, male, 6 years old, was born in Curitiba (PR), Brazil and living in Guaratingueta (SP), Brazil, developed "common cold", bimodal diary fever, chills, shake and sweats . Dyspnea, cough with general lymphadenopathy . Foot and right shoulder arthralgias . Six months ago visited a cave, equitation practice, dog and cat contacts and no transfusion, frontal sweats, fever (38.4 degrees C) . T.A . was 8/6, tachycardia in generalized lymphadenopathy . Cardiopulmonary system was normal, mesogastric tumoral mass, hepatosplenomegaly and no ascites . Bone marrow with eosinophilia; nodule demonstrated presence of P . brasiliensis, hypoalbuminemia; hyperglobulinemia; anemia; leukocytosis with eosinophilia . Immunodiffusion with exoantigen 43 kd of P . brasiliensis was 1/32 . Primary complex like Ghon was observed in interstitial pneumonia followed by mediastinal and mesogastric mass (35 to 40 days) . Clavicular osteolytic lesions (45 to 60 days) appeared during paracoccidioidomycosis therapy . Recovery was observed 2 months after treatment of acute infantile paracoccidioidomycosis.

Rev Soc Bras Med Trop, 1992 Jul-Sep, 25(3), 171 - 5
{The clinical parameters relevant for the differential diagnosis between mucocutaneous leishmaniasis and paracoccidioidomycosis}; Colombo AL et al.; We retrospectively evaluated the medical records of 15 patients diagnosed with leishmaniasis and of 28 patients diagnosed with paracoccidioidomycosis presenting with involvement of the oral-nasal-pharyngeal mucosa seen at Escola Paulista de Medicina from 1986 through 1990 . These patients were compared in regard to the following variables: sex, age, time since disease onset, location of lesion, and clinical complaints . Sex, time since disease onset, lesion at the nasal septum, palate, tongue, and lips, and history of oral pain, dysphagia/odynophagia, and nasal obstruction were significantly different in the two groups of patients . The authors point out to the relevance of these findings in the differential clinical diagnosis of leishmaniasis and paracoccidioidomycosis.

Clin Endocrinol (Oxf), 1992 Jun, 36(6), 545 - 51
Adrenocortical dysfunction in paracoccidioidomycosis: comparison between plasma beta-lipotrophin/adrenocorticotrophin levels and adrenocortical tests; Moreira AC et al.; OBJECTIVE: Paracoccidioidomycosis is an important cause of Addison's disease in South America . We have carried out an extensive and prospective study on paracoccidioidomycosis comparing glucocorticoid, mineralocorticoid and androgen function with adrenal regulators, ACTH/beta-LPH and plasma renin activity (PRA) . PATIENTS AND METHODS: Forty-seven male patients with active paracoccidioidomycosis were studied consecutively together with 20 healthy controls . On day 1, plasma aldosterone and PRA levels were measured in blood samples obtained from patients in the supine and erect position . On day 2 at 0900 h, baseline plasma samples were taken for ACTH, beta-lipotrophin (beta-LPH), cortisol, corticosterone, aldosterone, androstenedione (delta 4-A) and dehydroepiandrosterone sulphate (DHEA-S) . ACTH 1-24 (250 micrograms) was given i.v . and blood samples for these steroid assays were taken at 1 and 2 hours . RESULTS: Five patients (10%) had Addison's disease with high basal plasma ACTH and beta-LPH, and low cortisol levels after the ACTH test . In the remaining 42 patients, baseline ACTH and beta-LPH levels and plasma cortisol after ACTH were within the normal range . A high percentage of patients presented with reduced corticosterone (21% of patients) and aldosterone (23%) secretion and increased PRA (31%) . Plasma delta 4-A (19%) and DHEA-S (50%) levels were also reduced . CONCLUSIONS: The frequency of Addison's disease among our patients with paracoccidioidomycosis was 10% . In addition, a subset of patients presented with adrenal dysfunction detected by mineralocorticoid or androgen tests . In parallel to pathological lesions a functional adaptation may occur during adrenal involvement in paracoccidioidomycosis.

Infect Immun, 1992 May, 60(5), 2096 - 100
Fate of conidia of Paracoccidioides brasiliensis after ingestion by resident macrophages or cytokine-treated macrophages; Cano LE et al.; Conidia ingested by resident macrophages had an enhanced percentage of transformation to yeast cells compared with those in culture medium without macrophages . The yeast cells subsequently grew intracellularly by budding . Macrophages treated with cytokines from antigen-stimulated spleen cells from immunized mice significantly inhibited transformation of ingested conidia.

Clin Exp Immunol, 1992 Apr, 88(1), 119 - 23
Specific recognition pattern of IgM and IgG antibodies produced in the course of experimental paracoccidioidomycosis; Vaz CA et al.; Specific IgM and IgG responses to Paracoccidioides brasiliensis produced in resistant and susceptible mice during experimental paracoccidioidomycosis were examined by the immunoblotting procedure . Sera from infected mice recognized 51 antigen bands with apparent molecular masses from 8 to 86 kD . Sixteen of these were defined as major antigen bands because of almost universal presence of antibodies to them, and their intense staining . All sera, including those from normal control mice, tested for both IgM and IgG antibody reacted with the major E antigen which appeared as a large diffuse band from 43 to 47 kD . Comparisons between resistant and susceptible mice showed some significant differences in IgM responses to many antigen bands . While IgG responses were quite similar for both strains, differences were apparent in the response to the antigens at 62 and 68 kD.

Eur J Epidemiol, 1992 Mar, 8(2), 206 - 10
Histoplasmosis in northwestern Argentina . Epidemiological survey of Chuscha and La Higuera in the province of Tucumán; van Gelderen de Komaid A et al.; The present work was undertaken to obtain epidemiological data on the extent and distribution of Histoplasma capsulatum infections in the Chuscha and La Higuera areas . Skin test surveys of the human population with histoplasmin and paracoccidioidin were carried out in 40% of the permanent population of both localities, between 3 and 88 years old . It was found that 53.57% of the population were histoplasmin reactors and 1.86% of the population were paracoccidioidin reactors.

Clin Infect Dis, 1992 Mar, 14 Suppl 1, S68 - 76
A Pan-American 5-year study of fluconazole therapy for deep mycoses in the immunocompetent host . Pan-American Study Group; Diaz M et al.; Eighty-eight immunocompetent patients with deep mycoses from eight countries were evaluated with the same protocol for efficacy of fluconazole monotherapy . Entry doses were raised from 100 to 400 mg as safety was shown in initial cohorts, and dosages up to 2,400 mg daily and durations up to 44 months were studied . Results were very similar in different countries . Twenty-seven of 28 evaluable patients with paracoccidioidomycosis, 13 of 19 with sporotrichosis, 14 of 16 with coccidioidomycosis, and eight of eight with histoplasmosis demonstrated objective responses to therapy, as did one patient each with zygomycosis and alternariosis . For these patients, relapses have been unusual thus far . In contrast, one patient with chromoblastomycosis responded but relapsed, and six did not respond; one patient with mycetoma responded but relapsed, and two did not respond . The drug was well tolerated by patients, including six who received intravenous therapy . In vitro susceptibility tests suggested that clinical response was correlated with susceptibility but that resistance did not preclude clinical response . Fluconazole therapy appears efficacious for several deep mycoses; dosages of greater than 200 mg daily may be needed for some diseases . The further evaluation of fluconazole for these entities is warranted.

Rev Inst Med Trop Sao Paulo, 1992 Mar-Apr, 34(2), 85 - 90
{Bone marrow involvement and eosinophilia in paracoccidioidomycosis}; Shikanai-Yasuda MA et al.; The authors described three acute paracoccidioidomycosis patients with bone marrow involvement . P . brasiliensis yeast forms were observed in bone marrow smears of all them, and in one case, culture also revealed fungus growth . The mononuclear phagocytic system involvement, the blood eosinophilia and the negative skin hypersensibility responses were emphasized in all of them, as well as the severity of the disease in one case, with disseminated bone lesions and 20.260 eosinophils/mm3 in peripheral blood . The authors discuss the possible role of eosinophil in the host-parasite interaction in paracoccidioidomycosis, suggesting that TH 2 subpopulation activation and increased IL 5 and GM-CSF secretions may be responsible by eosinophilia in the most severe case.

Rev Inst Med Trop Sao Paulo, 1992 Mar-Apr, 34(2), 171 - 6
Activity of two different triazoles in a murine model of paracoccidioidomycosis; Restrepo S et al.; A new orally absorbable triazole (Schering 39304) with a long serum half-life in man (60 hours), was tried in a murine model of progressive paracoccidioidomycosis and compared with itraconazole, another triazole which has proven effective in this mycosis . Only 15% of the infected, untreated mice survived while 53 to 75% of the animals receiving itraconazole survived . Mice treated with Schering 39304 exhibited higher (86-100%) survival rates . Statistically, the 5 mg/kg Sch 39304 was superior to the 50 mg/kg itraconazole dose . Lung cultures showed that 20 mg/kg/day of Sch achieved sterilization of the infectious foci . These results indicate that the new triazole will have a place in the treatment of paracoccidioidomycosis.

Rev Inst Med Trop Sao Paulo, 1992 Mar-Apr, 34(2), 107 - 15
{Thoracic radiologic aspects in paracoccidioidomycosis}; do Valle AC et al.; In this paper the authors analyse 159 radiographs from paracoccidioidomycosis patients seen at the Evandro Chagas Hospital/Fiocruz in the period between January 1960 to December 1988 . Twenty four cases (15.09%) of association with tuberculosis were observed; one with pneumoconiosis; one with aspergillosis, and two with carcinoma . Twenty cases were excluded from the radiologic analysis: in 8 of these the diagnosis of tuberculosis occurred concomitantly, and in 12 patients, lung fibrosis due to previous treatment for tuberculosis or paracoccidioidomycosis was present in the 139 remaining cases, the radiographic abnormalities encountered were grouped according to the predominance of lesions at the various lung sites, if alveolar or interstitial, according to Magalhaes' (1982) classification modified by the authors: infiltrate 55 cases (39.6%); mist 28 (20.1%); pneumonic 23 (16.6%); nodular 16 (11.5%); micronodular 10 (7.2%), and fibrotic 7 (5.0%) . In 113 cases it was possible to follow the regression of the pulmonary process radiologically . In 85 (75.2%) patients, regression took place within 6 months; in 17 (15.0%) cases between 7 and 12 months; in 4 (3.5%) between 13 and 24 months, and in 7 (6.1%) cases no changes in the radiographic pattern were noted.

Rev Hosp Clin Fac Med Sao Paulo, 1992 Mar-Apr, 47(2), 103 - 5
{Kidney failure as a complication of the treatment of pulmonary paracoccidioidomycosis}; Fiks IN et al.; Sulfadiazine is one of the drugs of choice in the treatment of paracoccidioidomycosis . Side effects are uncommon . Cutaneous and gastrointestinal reactions and rarely leucopenia and jaundice may occur . A patient on sulfadiazine 6 g daily exhibiting a good response is reported . On the 12th day, he presented an intense abdominal pain and reduced urinary output . An ultrasonography revealed diffuse calculosis in urinary tract . Peritoneal dialysis, urinary alkalinization and rehydration were instituted with complete restoration of renal function . This case illustrates the importance of an adequate hydratation of patients receiving sulfadiazine, a simple measure in the prevention of this uncommon complication which carries a high morbidity.

Rev Saude Publica, 1992 Feb, 26(1), 12 - 6
{The relationship between paracoccidioidomycosis and alcoholism}; Martinez R et al.; The relationship between alcoholism and paracoccidioidomycosis was evaluated by the case-control method . The alcohol consumption of 4 groups of patients was compared: 50 patients with chronic paracoccidioidomycosis, 20 patients with the acute or subacute form of this mycosis and their respective control groups of hospitalized patients, each case matched by sex and age . Between September 1986 and July 1988 the cases and their controls were interviewed by one and the same investigator using a questionnaire on drinking habits: quantity and type of beverage consumed, time of onset and frequency of use and whether they had manifested symptoms of inebriation or of alcohol dependence previously . As compared with control patients, the mean daily ingestion of alcohol in excess of 60 ml was more frequent in the chronic paracoccidioidomycosis group (50.0% x 30.0%) . These patients also preferred to drink sugar cane brandy more frequently (89.4% x 68.3%) . When the average daily consumption of ethyl alcohol exceeded 100 ml, most patients presented a recurrence of infection during or after antifungal therapy . In the acute-subacute paracoccidioidomycosis group, 64.3% of the patients reported inebriation on one or more occasions, versus 17.6% in the respective control group . The results suggest that alcoholism can be a predisposing factor to paracoccidioidomycosis and, probably, accounts for a worse prognosis for this infection.

J Med Vet Mycol, 1992, 30(2), 161 - 8
Killing of Paracoccidioides brasiliensis conidia by pulmonary macrophages and the effect of cytokines; Cano LE et al.; The ability of conidia, the infectious form of the dimorphic fungus Paracoccidioides brasiliensis, to be killed in vitro by murine pulmonary macrophages was studied . Mice were immunized by intravenous injection of killed conidia, which resulted in cellular immunity demonstrated by delayed type hypersensitivity in vivo and macrophage migration inhibition factor production in vitro . Resident pulmonary macrophages from non-immune mice were able to significantly kill the conidia (28%) . Such macrophages treated with supernatants (cytokines) from antigen-stimulated immune mononuclears had a markedly enhanced ability to kill conidia (73%) . These results show that activated pulmonary macrophages are potent killers of conidia of P . brasiliensis and that immune mononuclears play a role in activation of macrophages . Activated macrophages may be important for pulmonary defense against the initial stages of infection with this fungus.

J Med Vet Mycol, 1992, 30(1), 51 - 60
Mutants of Paracoccidioides brasiliensis strain IVIC Pb9 affected in dimorphism; San-Blas F et al.; Morphological mutants were isolated after nitrosoguanidine treatment of Paracoccidioides brasiliensis strain IVIC Pb9 . Two of these mutants, Pb257 and Pb258, developed a typical mycelia at 23 degrees C, however, the yeast cells which developed at 37 degrees C were indistinguishable from those of the parental strain . A third mutant, strain Pb267, was thermosensitive, grew as yeast-like cells at 23 degrees C, but was unable to survive at 37 degrees C . Morphological observations as well as serological and segregation tests confirmed that the mutant strains originated from P . brasiliensis . Cell wall chemical analyses of the mutant strains grown at 23 degrees C indicated the presence of alkali-soluble, acid-insoluble polysaccharides absent in the parental wild-type strain Pb9 grown under the same conditions . The phenotypes shown by the mutant strains may be related to deficiencies in the proper synthesis of cell wall components of the mycelial phase of this fungus.

J Med Vet Mycol, 1992, 30(3), 261 - 4
The source of the growth-promoting factor(s) affects the plating efficiency of Paracoccidioides brasiliensis; Singer-Vermes LM et al.; We studied the influence of the growth factor (GF) source, concentration and production time on the plating efficiency of Paracoccidioides brasiliensis yeast cells . The highest plating efficiencies were achieved when the GF was derived from a fast growing P . brasiliensis isolate which was not homologous to the plated samples.

J Med Vet Mycol, 1992, 30(3), 173 - 84
Development of pulmonary fibrosis in mice during infection with Paracoccidioides brasiliensis conidia; Restrepo S et al.; Pulmonary fibrosis is a feared sequelae of paracoccidioidomycosis . We sought to determine if mice exposed to Paracoccidioides brasiliensis conidia would develop pulmonary fibrosis . BALB/c mice were infected intranasally with P . brasiliensis conidia and sacrificed at regular intervals . One lung was sectioned for histopathology and sections were stained with haematoxylin and eosin, trichromic and argentic stains; the other lung was homogenized and cultured to determine the viability of the fungus . One week post-challenge, only small peribronchial foci were apparent . After 4 weeks, reticular fibres appeared disorganized and disrupted . Six to 8 weeks later peribronchial infiltrates were larger and appeared surrounded by reticular fibres; thick collagen I fibres were noticed in the infiltrated areas at this time . On weeks 10-12, infiltrates were confluent and reticular fibres were concentrated around the inflammatory foci; collagenization was apparent . Observations up to 16 weeks revealed diffuse involvement of the lung parenchyma with extensive collagenization . Lung cultures were always positive . We suggest that inhalation of P . brasiliensis conidia induces adverse lung responses leading to changes in the proportion of collagen fibres I and III.

J Med Vet Mycol, 1992, 30(4), 317 - 21
Activation of human complement system in paracoccidioidomycosis; Munk ME et al.; Plasma samples of 14 patients with paracoccidioidomycosis were analysed for components that represent activation of the complement system . Most patients (12/13) showed significant titres of complement-fixing antibodies and 14/14 had increased C4d/C4 ratios . There was no conclusive correlation between these two immunological indices, however . Factor B values of patients were similar to normal donors and fragment Ba was not detected in any of the patients . These results indicate a classical complement pathway activation in the plasma of patients with paracoccidioidomycosis.

Acta Cient Venez, 1992, 43(6), 355 - 9
{Characterization of Paracoccidioides brasiliensis antigens . Serological immunodiagnosis using western blotting}; Ortega G et al.; The electrophoretic characterization on SDS-PAGE gels of Paracoccidioides brasiliensis antigens, strains 2511 and 6688; and the additional use of immunoblotting has permitted in this study to identify the immunogenic, sensitive and specific antigen fractions for the diagnosis of Paracoccidioidomycosis . The antigenic preparations showed differences depending on the morphologic form of the fungus and the method utilized . The procedure revealed heterogeneity in the humoral immune response of the patients studied and permitted us to establish indices of activity of Paracoccidioidomycosis by the sequential analysis of sera obtained during different stages of the disease . The high sensitivity of this method makes it useful as an additional technique for the serologic immunodiagnosis of Paracoccidioidomycosis.

Acta Cient Venez, 1992, 43(1), 3 - 10
{Biochemical regulations in the dimorphism and virulence of pathogenic fungi for humans}; San-Blas G; Some biochemical mechanisms involved in the processes of virulence and dimorphism in fungi pathogenic for humans are reviewed . Among them, the participation of sulphydryl and disulfide groups, hormone receptors and intra- and extracellular proteinases in Histoplasma capsulatum, Paracoccidioides brasiliensis and Coccidioides immitis.

Bull Soc Pathol Exot, 1992, 85(2), 118 - 9
{Keloid paracoccidioidomycosis assessed with gallium}; Braga FJ et al.; This rare mycosis is caused by Paracoccidiodis loboii and is found mainly among indians of the Amazon forest . It is a cutaneous disease . No visceral affection was ever described, though generalized skin disease has been reported . One case of cheloid paracoccidioidomycosis was studied with gallium-67 . The radiopharmaceutical has accumulated in the affected area and in a lymph node, although adenomegaly be rare . Degree of accumulation was discrete/moderate but the patient had been under irregular treatment for 10 years . As no laboratorial data are evaluable for this affection, gallium-67 may represent a means of disease extent and therapy efficacy evaluation.

Bull Soc Pathol Exot, 1992, 85(2), 115 - 7
{Paracoccidioidomycosis assessed with 4 isotopic studies}; Braga FJ et al.; Paracoccidioidomycosis (south-american blastomycosis) is a consumptive granulomatous disease that is found in all Latin-America . The ethiological agent is a fungus (Paracoccidiosis brasiliensis) and mechanisms of disease transmission are not clear . Affection is frequent in the lungs, liver, spleen, ganglia, oropharynx and brain . Disease's response to therapy (Amphotericin, Ketoconazole) is good but recurrence is frequent . We have studied 12 patients with gallium-67, bone scintigraphy, bone marrow scintigraphy and lymph-scintigraphy . Six of these patients were re-studied after variable time of therapy . Gallium-67 accumulated in all patients's lungs . Other abnormal areas included ganglia, liver, spleen and bone . Bone scintigraphy was abnormal in all bone sites that were abnormal at the gallium study . Bone marrow scintigraphy showed expansion to the periphery in 6/12 cases and lymph nodes accumulated the radiopharmaceutical in 5 patients . Patients re-studied after the beginning of therapy presented decreased degrees or normalization of the radio-pharmaceutical's uptake (gallium and MDP), a regression of the bone marrow periphery expansion (bone marrow scintigraphy) and normalization of the lymph node aspects . These four radioisotopical studies may be useful for staging and for therapy follow-up . The sensitivity of the method is greater than the one noted for the conventional radiological studies.

Chemotherapy, 1992, 38 Suppl 1, 27 - 34
Tropical mycoses; Bayles MA; The most common tropical subcutaneous and deep mycoses include chromomycosis, sporotrichosis and mycetoma . All are commonly found in Natal and in other subtropical countries . Although blastomycosis is endemic in North America, only 4 cases have been identified in Natal during the last 25 years, and all presented with atypical clinical features . African histoplasmosis, caused by Histoplasma capsulatum var . duboisii and limited mainly to central and western Africa, has been found in only 1 patient in Natal . Paracoccidioidomycosis, though the most common deep mycosis in Latin America, is limited to that area and there is no experience of this disease in South Africa . Over the past 8 years, itraconazole has been used in clinical trials for all 3 mycoses . The results in sporotrichosis, non-meningeal blastomycosis and paracoccidioidomycosis suggest that for these diseases itraconazole may be the drug of choice . The results in histoplasmosis are encouraging, as are the results in chromomycosis, particularly those cases associated with Cladosporium carrionii . Where Fonsecaea pedrosoi is the causal agent and in mycetomas, however, successful management still remains a therapeutic problem . Enhanced efficacy by combining flucytosine and itraconazole was seen in 3 patients . Even over prolonged periods, itraconazole has an impressive safety profile . In the present series of 41 patients, no side-effects were observed, no adverse reactions occurred, and serum chemistry values remained within normal limits . It appears, therefore, that itraconazole, though not the final answer to management of deep mycoses, is certainly a major improvement on previous drugs.

J Med Vet Mycol, 1992, 30(6), 481 - 4
Activation of human complement system Paracoccidioides brasiliensis and its deposition on the yeast form cell surface; Munk ME et al.; The yeast form of Paracoccidioides brasiliensis strain Pb18 was able to activate C3 of normal human serum diluted in phosphate-buffered saline or EGTA-MgCl2 in vitro . C3 convertase was also permissive when Pb18 cells were pre-treated with a pool of immune serum from patients with paracoccidioidomycosis and incubated in serum diluted in EDTA-CaCl2 . The components C3, and fragments C3c, C3d, C3g, factor H, factor B, C4 and C5b-9 were demonstrated on the Pb18 cell surface by immunofluorescence although no effect was seen on fungal viability.

Semin Respir Infect, 1991 Dec, 6(4), 235 - 46
Pulmonary infections in immigrants and refugees; Iralu JV et al.; Immigrants and refugees are susceptible to a wide array of pulmonary diseases, including familiar infections that are acquired after entry and unusual infections that are imported from abroad . The challenges are great; tuberculosis (TB) is now 13 times more prevalent among immigrants than among the general US population . Melioidosis, paracoccidioidomycosis, and paragonimiasis are often mistaken for TB . Pulmonary disease caused by parasites is uncommon and particularly perplexing unless a peripheral blood eosinophilia suggests helminthic infection . TB, melioidosis, fungal infections, and strongyloidiasis may remain inapparent for years and then produce devastating illness in the setting of immunosuppressive therapy or the acquired immunodeficiency syndrome . Proper health care of immigrants requires an understanding of unusual exposures and infections and should include preventive measures as well.

Med Klin (Munich), 1991 Dec, 86 Suppl 1, 13 - 9
{Tropical mycoses}; Bayles MA; The most common tropical subcutaneous and deep mycoses include chromomycosis, sporotrichosis and mycetoma . All are commonly found in Natal and in other sub-tropical countries . Although blastomycosis is endemic in North America, only four cases have been identified in Natal during the last 25 years and all presented with atypical clinical features . African histoplasmosis caused by H . capsulatum var . duboisii and limited mainly to central and western Africa has been found in only one patient in Natal . Paracoccidioidomycosis, although the most common deep mycosis in Latin America, is limited to that area and we have no experience of this disease in South Africa . Over the past eight years itraconazole has been used in clinical trials for all these mycoses . The results in sporotrichosis, non-meningeal blastomycosis and paracoccidioidomycosis suggest that for these diseases itraconazole may be the drug of choice . The results in histoplasmosis are encouraging as are the results in chromomycosis particularly those cases associated with C . carrionii . However, where F . pedrosoi is the causal agent and in mycetomas, successful management still remains a therapeutic problem . In our own experience and that of other using itraconazole, even over prolonged periods, this drug has an impressive safety profile . In our present series of 42 patients, no side effects were observed, no adverse reactions occurred and serum chemistry values remained within normal limits . It appears, therefore, that itraconazole, although not the final answer to management of the deep mycoses, is certainly a major improvement on previous drugs.

Rev Inst Med Trop Sao Paulo, 1991 Nov-Dec, 33(6), 503 - 8
Comparison on the performance of Leishmania major-like and Leishmania braziliensis braziliensis as antigen for New World leishmaniasis IgG-immunofluorescence test; Guimaraes MC et al.; The performance of an antigen of L . major-like promastigotes for the serological diagnosis of mucocutaneous leishmaniasis in the IgG-immunofluorescent test was compared to that of an antigen of L.braziliensis braziliensis . Each antigen was used to test two hundred and twenty-four sera of etiologies such as mucocutaneous leishmaniasis, deep mycoses, toxoplasmosis, malaria . Chagas' disease, visceral leishmaniasis, anti-nuclear factor, schistosomiasis, rheumatoid factor and normal controls . Agreement between responses to each antigen was high: 77.2% of leishmaniases sera agreed on a positive or a negative result to both antigens and 91.1% of control sera . Cross reactivity was restricted to Chagas' disease sera, visceral leishmaniasis, anti-nuclear factor and paracoccidioidomycosis . The quantitative response of leishmaniasis and Chagas' disease sera to both antigens was evaluated by a linear regression; although the y-intercept and the slope were different for each antigen, neither was better than the other in the disclosure of anti-Leishmania antibodies . In the case of Chagas' disease sera the L.major-like antigen was better than L.b.braziliensis' to disclose cross-reacting antibodies.

Mycoses, 1991 Nov-Dec, 34(11-12), 493 - 7
Paracoccidioidomycosis in Quillabamba, Peru, and its surroundings; Garcia R et al.; Three cases of paracoccidioidomycosis observed in 1990-1991 in Quillabamba, Peru, and its surroundings are described . The climate and the living conditions of the inhabitants favour paracoccidioidomycosis . It can be assumed that a majority of cases could not be identified, because the symptoms are disguised as other tropical diseases . The application of ketoconazole to the patients showed different results, from optimal to scarce, while temporary remission of the disease was obtained with itraconazole . The best therapy for paracoccidioidomycosis appears to be the application of amphotericin B plus sulphamidics.

Oral Surg Oral Med Oral Pathol, 1991 Oct, 72(4), 430 - 5
Oral manifestations of paracoccidioidomycosis (South American blastomycosis); de Almeida OP et al.; Paracoccidioidomycosis (South American blastomycosis) is an uncommon, progressive systemic mycosis, potentially fatal if untreated . It is virtually restricted to persons spending time in Latin America . Reports of oral lesions are extremely rare in the English-language literature . Three adults with oral lesions as the first sign of paracoccidioidomycosis are described; this appears to be the largest series in the dental literature . The oral lesions had a characteristic appearance with a granular purpuric surface . The upper gingiva was a typical site, but lesions were also seen in the palate, tongue, and buccal mucosa . Two of the patients proved to have detectable pulmonary involvement . Long-term systemic ketoconazole therapy produced resolution of oral lesions in all cases.

Rev Soc Bras Med Trop, 1991 Oct-Dec, 24(4), 253 - 5
{Calcified abdominal lymph node paracoccidioidomycosis}; da Silva AL et al.; The calcified form of the South-american blastomycosis is rarely mentioned in the literature . An observation of intra-abdominal lymphatic and visceral spread with abdominal calcification is accompanied by a radiological record . Are done comments . The pathogenesis and rarity of this complication is discussed.

Arch Biochem Biophys, 1991 Sep, 289(2), 298 - 302
The 43-kDa glycoprotein from the human pathogen Paracoccidioides brasiliensis and its deglycosylated form: excretion and susceptibility to proteolysis; Puccia R et al.; Biochemical properties of the concanavalin A-binding 43-kDa glycoprotein (gp43) of Paracoccidioides brasiliensis and its deglycosylated form were compared . Deglycosylation was achieved by treatment with trifluoromethanesulfonic acid, endoglycosidase H, N-glycanase, or metabolically, by growing cells with tunicamycin . The resulting antigen in all cases had Mr 38,000, and probably derived from the gp43 by loss of N-linked high-mannose oligosaccharide chains . The presence of galactopyranose units in the carbohydrate chains was suggested by antigen binding to peanut lectin . Pulse and chase experiments using {35S}methionine metabolic labeling of P . brasiliensis growing in the presence of tunicamycin showed that the N-linked chains of gp43 are not required for antigen secretion . The 38-kDa antigen was more susceptible than the native antigen to the action of papain and pronase, thus indicating a protective role of the carbohydrate moiety against proteolysis . Both forms are equally resistant to endogenous proteases at neutral pH . The gp43, itself, has a proteolytic activity at pH 5-6, but not at neutral pH . Deglycosylation with endoglycosidase H or tunicamycin preserved epitopes in the 38-kDa molecule reactive with (a) antibodies from patients with paracoccidioidomycosis, or rabbit immunized with the gp43 and (b) mouse monoclonal antibodies against the gp43 antigen . The present results provide a basis for the understanding of diagnostic reactions and fungal virulence involving the gp43 exocellular antigen of P . brasiliensis.

Rev Inst Med Trop Sao Paulo, 1991 Sep-Oct, 33(5), 407 - 11
{Paracoccidioidomycosis among the indian group Suruí of Rondonia, Amazonia, Brazil . A case report}; do Valle AC et al.; The Authors report a case of paracoccidioidomycosis in a Surui Indian patient from the state of Rondonia, Brazilian Amazon . The subject is an adult male, having been diagnosed on the basis of mycologic, serologic, and radiographic exams . The prescribed therapy was sulphamethoxazole 800 mg associated with trimethoprim 160 mg every 12 hours . A reevaluation of the patient conducted six months after the beginning of chemotherapy indicated overall improvement of his physical condition and of the radiologic picture, negativeness of the test of precipitin in capillary tube, and positiveness of the skin test with paracoccidioidin . The Authors also reviewed the cases of paracoccidioidomycosis described in the region, especially among the Surui Indian population.

Rev Hosp Clin Fac Med Sao Paulo, 1991 Sep-Oct, 46(5), 219 - 22
{Incidence of adrenalitis in necropsy material}; Fernandes VS et al.; In the necropsy material of the Medical School of the University of Sao Paulo, collected between 1931 and 1981, consisting of 131,466 post-mortem examinations, there were 254 (0.2%) cases of adrenalitis, 185 male and 69 female . In 161 of these cases the affection was bilateral, in 46 it was unilateral and in 47 cases this information is lacking . In 111 cases (43.7%), the adrenalitis was caused by tuberculosis, in 86 cases (33.8%), by South American blastomycosis, in three cases (1.2%) by histoplasmosis, in two (0.8%) by Salmonellosis, in two (0.8%) by cytomegalovirus infection and in one (0.4%) by leishmaniasis . The high number of cases with tuberculosis and paracoccidioidomycosis of adrenal glands is due to high incidence of these infections in Brazil.

Mycopathologia, 1991 Aug, 115(2), 73 - 81
Studies on the relationship between paracoccidioidomycosis in ddY mice and their estrous cycle; Sano A et al.; The relationship between paracoccidioidomycosis in ddY mouse and its estrous cycle was studied . Adult ddY mice of both sexes were used as experimental animals . Estrous cycle of female mice was examined before inoculation of Paracoccidioides brasiliensis yeast cells and mice were divided into 5 groups such as proestrus, estrus, metestrus-I, metestrus-II and diestrus . Each mouse was inoculated intravenously with 10(6) P . brasiliensis yeast cell units and sacrificed on day 28 after inoculation . Their internal organs were cultured, and in addition, their histopathologies were studied . As a result, there was no difference in the organ cultures among the male and the female mice of 5 groups . However, histopathologically, the female groups at estrus, metestrus-I and metestrus-II were affected more severely than the male group, and the susceptibility of the female mice to the fungus was closely related to their estrous cycles.

J Clin Microbiol, 1991 Aug, 29(8), 1610 - 5
43-kilodalton glycoprotein from Paracoccidioides brasiliensis: immunochemical reactions with sera from patients with paracoccidioidomycosis, histoplasmosis, or Jorge Lobo's disease; Puccia R et al.; Sera from patients with paracoccidioidomycosis (PCM), histoplasmosis (HP), or Jorge Lobo's disease (JL) were titrated against purified gp43 from Paracoccidioides brasiliensis by using both enzyme-linked immunosorbent assay (ELISA) and immunoprecipitation (IPP) reactions with 125I-labeled antigens . In IPP, PCM sera and other sera could be distinguished on the basis of serum titers, whereas in ELISA, 53% of the HP sera and 29% of the JL sera reacted similarly to the PCM sera . To investigate the possible role of the carbohydrate epitopes in these reactions, we compared the reactivities of sera from several patients with native and deglycosylated gp43 . Competition experiments were carried out with monosaccharides as inhibitors . The results suggest that greater than 85% of the reactions of the PCM sera with gp43 involved peptide epitopes . Cross-reactions with HP and JL sera in ELISA were predominantly attributed to periodate-sensitive carbohydrate epitopes containing galactosyl residues . HP and JL sera which reacted strongly with gp43 in ELISA were only weakly reactive or did not react in IPP with labeled antigens in solution . Moreover, ELISA reactions could be significantly inhibited either by monosaccharides or by periodate treatment . Apparently, carbohydrate epitopes in gp43 are more accessible to the antibodies when the molecule is bound to a plastic substrate than when it is in solution . Structural changes in the gp43 antigen arising by N deglycosylation abolish reactivity with PCM sera and support the existence of conformational peptide epitopes.

Rev Inst Med Trop Sao Paulo, 1991 Jul-Aug, 33(4), 319 - 24
{Utilization of aminoacids in the study of the growth of Paracoccidioides brasiliensis . Influence on dimorphism}; Cano MI et al.; Fifteen Paracoccidioides brasiliensis strains, in the mycelial (M) and yeast like (Y), were cultivated in minimal medium (MM) and subcultivated to be adapted to the same medium supplemented with a pool of amino acid in solution (MMS) . Each of the amino acids were studied separately of the solution to provide the auxological study . The prototrophism was demonstrated by nine strains in both M and Y forms, and the auxotrophism by the remaining strains . The heterologous results has not allowed us to draw an auxological characterization of the P . brasiliensis . As far as we could observe none of the amino acid studied in this piece of research can be considered of absolute importance for to the growth and the morphogenesis maintainance of the fungus . Morphological alterations were only verified in the prototroph strains, which suggest that there could have been adaptative metabolism activity due to the absence of organic nitrogen compounds in the minimal medium (MM).

Rev Inst Med Trop Sao Paulo, 1991 Jul-Aug, 33(4), 277 - 80
The sensitivity, specificity and efficiency values of some serological tests used in the diagnosis of paracoccidioidomycosis; Del Negro GM et al.; This work reports on the results of double immunodiffusion (ID), counterimmunoelectrophoresis (CIE), complement fixation (CF) and indirect immunofluorescence (IIF) techniques in the serodiagnosis of paracoccidioidomycosis . The study was undertaken on four groups of individuals: 46 patients with untreated paracoccidioidomycosis, 22 patients with other deep mycoses, 30 with other infectious diseases (tuberculosis and cutaneous leishmaniasis) and 47 blood donors as negative controls . Data were obtained using Paracoccidioides brasiliensis antigens, i.e., a yeast culture filtrate for ID, CIE and CF, and a yeast cell suspension for IIF . The sensitivity, specificity and efficiency values were measured according to GALEN & GAMBINO . The gel precipitation tests (ID and CIE) showed the greatest sensitivity (91.3 and 95.6%, respectively), maximum specificity (100%) and the highest efficiency values when compared to the CF and IIF tests.

Rev Inst Med Trop Sao Paulo, 1991 Jul-Aug, 33(4), 267 - 76
{Paracoccidioidomycosis: radiologic and pulmonary study in 58 cases}; Campos EP et al.; Spirometric and haemogasimetric tests were performed in 52 patients with unifocal and multifocal progressive Paracoccidioidomycosis pulmonary diseases, in 5 patients with juvenile and 1 female internal genital forms . The age ranged from 20 to 74 years old and the duration of disease from 3 to 25 years . Dyspnea, cough, tegumentary and lymphatic manifestations were the most predominant signs observed . Fourteen of them was developed Cor pulmonale . Impaired renal function was recorded in 19 patients, cardiac symptoms were described in 7 and hyporeactor adrenal activity was showed in 13 patients . Radiological studies demonstrated mild in 16, moderate in 24 and severe pulmonary lesions types in 18 patients . The pulmonary function revealed: normal spyrographic findings in 17, pure obstructive type in 32 and mixed form in 9 of them . Hyperventilation was described in 54 individuals and all of them showed an increasing of the alveolo-arterial difference . PaO2 less than 80 mm/Hg observed in 36 of them . Statistical analysis demonstrated significative association between clinical evolution and radiological interpretation . Similar data were obtained in radiology evaluations, clinical evolutive studies and pulmonary functions described in these patients . The granulomatous reaction due to Paracoccidioidomycosis, in heavy smokers patients, gave origin to the alterations in small airways predisposing the interalveolar dissemination an impaired alveolo-arterial diffusion.

Histol Histopathol, 1991 Jul, 6(3), 395 - 402
Leukocyte immunophenotypes in bronchoalveolar lavage fluid and peripheral blood of paracoccidioidomycosis, sarcoidosis and silicosis; Tapia FJ et al.; Leukocyte subsets in bronchoalveolar lavage (BAL) fluid and peripheral blood of patients with paraccoccidioidomycosis, sarcoidosis and silicosis were characterized using monoclonal antibodies and an immunoperoxidase technique . In paraccocidioidomycosis, the number of T-helper/inducer CD4-positive lymphocytes was lower in peripheral blood than in BAL fluid . Additional analysis showed that the expression of HLA-DR was very similar in alveolar macrophages, lung and blood T-cells . In sarcoidosis and silicosis there were higher proportions of T-helper/inducer cells in peripheral blood than in BAL fluid . The alterations in the T-helper/inducer/T-suppressor/cytoxic CD4/CD8 ratio in sarcoidosis and silicosis were more appreciable in peripheral blood than in BAL fluid, contrasting with the results in paracoccidioidomycosis . The expression of HLA-DR by alveolar macrophages in sarcoidosis was the highest of all the disease studied . No statistically significant differences were observed between chronic multifocal and chronic unifocal paracoccidioidomycosis disease, stage II and stage III sarcoidosis, and chronic and accelerated silicosis . The three granulomatous diseases analyzed had a few alveolar macrophages expressing the CD4 molecule on their surface . These findings and the technique of analyzing both peripheral blood and BAL leukocyte subsets may help to understand the pathogenesis of interstitial lung diseases.

J Clin Microbiol, 1991 Jun, 29(6), 1202 - 5
Sensitive immunoradiometric assay for the detection of Paracoccidioides brasiliensis antigens in human sera; Ferreira-da-Cruz MF et al.; In the present study we report the standardization of an immunoradiometric assay (IRMA) for detection of Paracoccidioides brasiliensis circulating antigens that could be useful in the diagnosis and prognosis of paracoccidioidomycosis . For this purpose we studied the reactivities of P . brasiliensis and other mycotic antigens with rabbit polyclonal anti-P . brasiliensis antibodies (immunoglobulin G) in order to evaluate the sensitivity and specificity of an IRMA for detecting P . brasiliensis antigens . The results were compared with those obtained by the double immunodiffusion test, the standard technique for the serodiagnosis of paracoccidioidomycosis . By using the immunoglobulin G fraction of rabbit antisera (900 ng per well), it was possible to detect up to 3.6 ng (0.12 micrograms/ml) of cellular antigen and 360 ng (12 micrograms/ml) of metabolic antigen in contrast to the double immunodiffusion test that could detect only 12 micrograms (1.2 mg/ml) of both antigens . IRMA was shown to be feasible and very sensitive and may therefore help, together with clinical data, in establishing early diagnosis and assessing disease activity . It could also allow the study of relationships between P . brasiliensis circulating antigens and host defense mechanisms during the disease.

Mycopathologia, 1991 Jun, 114(3), 187 - 91
Sputum cytology in the diagnosis of pulmonary paracoccidioidomycosis; de Mattos MC et al.; The presence of Paracoccidioides brasiliensis was determined in sputum samples from 50 patients with paracoccidioidomycosis using four different techniques: (a) cell-block preparations stained with silver methenamine, (b) direct microbiologic examination, (c) smears stained with Shorr, and (d) smears stained with silver methenamine . Overall, cell-block preparations and smears stained with silver methenamine proved to be the most sensitive techniques, followed by smears stained with Shorr and direct microbiologic examination in decreasing order of sensitivity . Sputum cytology tended to be less positive in patients with interstitial pulmonary lesions as determined by chest X-ray than in patients with alveolar lesions . In addition to its high sensitivity, cell-block preparation technique allows storage of blocks and slides for further studies.

Mycopathologia, 1991 Jun, 114(3), 169 - 77
Electron microscopic study of conidia produced by the mycelium of Paracoccidioides brasiliensis; Edwards MR et al.; The ultrastructure of asexual spores (conidia) produced by the mycelial form of Paracoccidioides brasiliensis was studied for the first time with transmission electron microscopy, using thin sections of aldehyde-osmium-fixed and epoxy-resin-embedded samples . The various types of conidia observed in the sections correlated well with previous light-microscopic descriptions . These types were intercalary or apical conidia, depending on their location along the originating hyphae . As in previous studies they were characterized as arthroconidia, aleuriospores and sessile or pedunculate pyriform conidia . The sporogenous cells were clearly distinguished from hyphal cells by the thickness and appearance of their cell walls . Copious fibrillar material (glycocalyx) detected at the cell surface was stained with ruthenium red during the fixation process . Typical subcellular organelles (nucleus, nucleolus, mitochondria, ribosomes, etc) were found in most of the sections . It was concluded that the spores produced by the mycelial phase of P . brasiliensis possess all attributes of viable and physiologically competent eukaryotic cells.

J Neurol Sci, 1991 Jun, 103(2), 179 - 81
Tapia's syndrome caused by Paracoccidioidis brasiliensis; de Freitas MR et al.; Tapia's syndrome is due to extracranial involvement of the XIIth nerve and the recurrent laryngeal branch of the Xth nerve . There is ipsilateral paralysis of vocal cords, soft palate and tongue . The main causes are parotid and other tumors or injuries to the high neck . We describe here a Brazilian female patient who presented with a lesion in the nasal mucosa, and soon after had dysphagia and dysphonia . Examination revealed paralysis of the soft palate, vocal cords and tongue ipsilaterally . Microscopic examination of the lesion in the nasal mucosa revealed the presence of the fungus . The patient was treated with sulfonamide and ketoconasol.

Arq Neuropsiquiatr, 1991 Jun, 49(2), 192 - 7
{Paracoccidioidomycosis of intramedullar and cerebral localization}; Morato-Fernandez RN et al.; It is reported our diagnosis and therapeutical experience with a patient that presented clinical, radiological and tomographical manifestations of paracoccidioidomycosis with involvement of lungs, brain and spinal cord . Besides being a rare case the authors call attention for the extreme difficulty in defining the final diagnosis, which was achieved only after laminectomy and surgical biopsy of the intra spinal cord tumor at the C5 level . The patient received two series of treatment with ketoconazole (400-600 mg/day) . After both the patient presented tonic and clonic convulsive disorders . We speculated about a possible perigranulomatous inflammatory response related to the therapeutical intervention, not yet described in literature.

J Investig Allergol Clin Immunol, 1991 Jun, 1(3), 185 - 93
Suppression of antibody response to an unrelated antigen in experimental murine paracoccidioidomycosis: effect of cyclophosphamide and indomethacin; Amarante-Mendes JG et al.; A suppressive effect of experimental paracoccidioidomycosis on the IgE antibody response to an unrelated antigen (ovalbumin) has been previously observed in mice . This effect was restricted to a short period, reaching maximum levels when OA was administered on the third day of Pb-infection . In order to study possible mechanisms involved in the establishment of this suppression, resistant (A/SN) and susceptible (B10.A) mice were treated with either a low dose of cyclophosphamide (CY) or indomethacin (INDO), a potent inhibitor of prostaglandin synthesis . While treatment with the first drug in A/SN mice induced only a recovery of the IgE anti-OA antibody response, in B10.A mice this effect was extended to IgG1, IgG2a and total levels of anti-OA antibodies . On the other hand, treatment with INDO reverted the anti-OA antibody suppression regarding each antibody class tested and in both strains of mice . These results suggest the participation of prostaglandins and of a cyclophosphamide-sensitive mechanism in the induction of the suppressive phenomenon in the human system.

Pathologica, 1991 May-Jun, 83(1085), 311 - 6
Fine needle aspiration cytology in the diagnosis of non-neoplastic diseases of head and neck masses; Oyafuso MS et al.; In this study, 3 patients with head and neck masses, which were suspicious of neoplastic compromise, were submitted to FNAC investigation . Two cases showed specific infection by Paracoccidioidomycosis and one by Actinomycosis . The cytologic findings were in agreement with histopathologic diagnosis and clinical follow-up; that results showes that FNAC importance in the diagnosis of non-neoplastic masses could be explored in ambulatorial routine.

Mycopathologia, 1991 May, 114(2), 89 - 91
HLA antigens in Brazilian patients with paracoccidioidomycosis; Goldani LZ et al.; Eighty patients with paracoccidioidomycosis were typed for 43 HLA specificities from loci A, B, C and DR . A highly significant increased frequency of HLA-B40 (relative risk 29.2) and HLA-Cw1 (relative risk 8.8) were found in patients compared to control subjects . The frequencies HLA-A2, B7 and B21 were also increased in patients and haplotypes-B40-Cw1 and -A2-B40 were positively correlated with the disease . DR antigen frequencies were not significantly altered in the patients and evidence of a protective effect was not found for any of the 43 antigens tested . These findings further support the involvement of the HLA system in the genetic susceptibility to paracoccidioidomycosis and the importance of ethnic variability in this association.

Mycopathologia, 1991 May, 114(2), 93 - 101
Studies on the relationship between the pathogenicity of Paracoccidioides brasiliensis in mice and its growth rate under different oxygen atmospheres; Sano A et al.; We performed comparative studies of the pathogenicity of six strains of Paracoccidioides brasiliensis (Bt-9, Bt-4, Pb-9, Pb-18, Bt-7 and B-1183) for young adult male ddY mice and the growth rate of each strain under different oxygen atmospheres (aerobic, micro-aerobic and anaerobic atmospheres) at 37 degrees C . 10(6) units of yeast cells were intravenously injected into each mouse . The pathogenicity of each isolate was determined by a scoring system based on organ culture and histopathological findings . The growth rates under different oxygen atmospheres were determined by a scoring system in which 300 fungal units per strain were counted . The strain Bt-9 showed the greatest pathogenicity, followed by Bt-4, Pb-9 and Pb-18 had on intermediate rank of pathogenicity . Bt-7 and B-1183 were the least pathogenic of the strains tested . Except for strain Bt-7 all strains showed an excellent growth under an aerobic atmosphere . Bt-4 and Bt-9 also showed excellent growth under a micro-aerobic atmosphere, followed by Pb-9, whereas the growth of Pb-18, Bt-7 and B-1183 was limited . There was a correlation between the growth rate under a micro-aerobic atmosphere and the pathogenicity of a strain . The growth rate of P . brasiliensis under a micro-aerobic atmosphere strongly correlated to its pathogenicity.

Mycopathologia, 1991 Apr, 114(1), 9 - 15
Scanning electron microscopy of the conidia produced by the mycelial form of Paracoccidioides brasiliensis; Samsonoff WA et al.; The conidia produced by the mycelial form of Paracoccidioides brasiliensis were examined by scanning electron microscopy for the first time . Several different conidial types were characterized . These included intercalary arthroconidia, several types of septate conidia that are formed from other conidia, pedunculate conidia, and terminal hyphal conidia . In addition, the ultrastructure of the supporting pedestal of the pedunculate conidium was found to be separated from the mother conidium by a septum in some instances, and at other times it was not.

Rev Soc Bras Med Trop, 1991 Apr-Jun, 24(2), 111 - 4
Isoniazid acetylating phenotype in patients with paracoccidioidomycosis and its relationship with serum sulfadoxin levels, glucose-6-phosphate dehydrogenase and glutathione reductase activities; Barraviera B et al.; The authors evaluated the isoniazid acetylating phenotype and measured hematocrit, hemoglobin, glucose-6-phosphate dehydrogenase and glutathione reductase activities plus serum sulfadoxin levels in 39 patients with paracoccidioidomycosis (33 males and 6 females) aged 17 to 58 years . Twenty one (53.84%) of the patients presented a slow acetylating phenotype and 18(46.16%) a fast acetylating phenotype . Glucose-6-phosphate-dehydrogenase (G6PD) activity was decreased in 5(23.80%) slow acetylators and in 4(22.22%) fast acetylators . Glutathione reductase activity was decreased in 14(66.66%) slow acetylators and in 12(66.66%) fast acetylators . Serum levels of free and total sulfadoxin were higher in slow acetylator (p less than 0.02) . Analysis of the results permitted us to conclude that serum sulfadoxin levels are related to the acetylator phenotype . Furthermore, sulfadoxin levels were always above 50 micrograms/ml, a value considered therapeutic . Glutathione reductase deficiency observed in 66% of patients may be related to the intestinal malabsorption of nutrients, among them riboflavin, a FAD precursor vitamin, in patients with paracoceidioidomycosis.

Mycoses, 1991 Mar-Apr, 34(3-4), 133 - 40
Efficiency of crude and purified fungal antigens in serodiagnosis to discriminate mycotic from other respiratory diseases; Toriello C et al.; Mycotic immunodiagnosis was performed in 186 hospitalized patients with different respiratory diseases, mostly considered as tuberculosis and others with a doubtful diagnosis . Crude histoplasmin, coccidioidin, paracoccidioidin, blastomycin, candidin, aspergillin, and sporotrichin, as well as purified polysaccharide-protein complexes (PPC) of Histoplasma capsulatum, Coccidioides immitis, and Paracoccidioides brasiliensis were used as antigens . Immune tests used included skin test (ST), gel immunodiffusion (ID), counterimmunoelectrophoresis (CIE), complement fixation (CF), and ELISA . A possible association with candidosis was observed in 17% of patients with tuberculosis and diabetes; one presumptive paracoccidioidomycosis, one confirmed aspergillosis, and six cases of active histoplasmosis were determined . Candidin ST showed 29% of positive reactions with an increased frequency in patients between 31 and 55 years of age . CF test showed the highest positivity percentages with crude antigens, specially for Candida antigen (26.3%) and histoplasmin (18.2%) . Cross reactions were evident with crude antigens but decreased when PPC's were used in ELISA.

J Med Vet Mycol, 1991, 29(1), 57 - 60
Purification of the 43 kDa glycoprotein from exocellular components excreted by Paracoccidioides brasiliensis in liquid culture (TOM medium); Puccia R et al.; The 43 kDa glycoprotein (gp43) antigen has been purified by affinity chromatography and Sephacryl S-200 gel filtration from concentrated and dialysed culture supernatant fluids of Paracoccidioides brasiliensis grown in a tomato juice-enriched complex medium (TOM) . In the TOM medium, there was rapid and extensive growth of the fungus, with an abundant production of gp43 in relation to other exocellularly accumulated components . The purified gp43 can be used for immunochemical and structure determination studies.

J Med Vet Mycol, 1991, 29(5), 331 - 4
The use of a mixture of somatic and culture filtrate antigens in the evaluation of the immune response to Paracoccidioides brasiliensis; da Silva MI et al.; Three Paracoccidioides brasiliensis antigens, namely a culture filtrate preparation, a somatic antigen and a mixture of equal parts of the two, were tested by two serological techniques against sera from patients with paracoccidioidomycosis, and in an in vivo delayed hypersensitivity model in mice . The antigen mixture was more sensitive than the two individual antigens for the evaluation of the humoral and cellular immune response to P . brasiliensis, both in man and in experimental animals.

J Med Vet Mycol, 1991, 29(1), 31 - 8
The use of cell-free antigens of Paracoccidioides brasiliensis in serological tests; Camargo ZP et al.; A simple and rapid method for extracting specific cell-free antigens (CFA) from the yeast form of Paracoccidioides brasiliensis grown on agar slants was developed . Extracts were analysed mainly by immunodiffusion (ID) tests, for the immunodiagnosis of paracoccidioidomycosis . Extracts were obtained from 2 to 7-day-old cultures . The extract obtained after the third day, which contained 200-300 micrograms of protein/ml, gave the best ID reactions, with 100% sensitivity and specificity . The main precipitation band was intense and showed total identity with that formed by the specific 43 kDa glycoprotein . Other tests such as counterimmunoelectrophoresis and immunoelectrophoresis were also used successfully with CFA . The 3-day CFA extract showed a pattern of polypeptides in the 10-110 kDa range after SDS-polyacrylamide gel electrophoresis . Five components of 20, 43, 45, 55 and 70 kDa reacted on immunoblots with sera from patients with active chronic paracoccidioidomycosis . The specific diagnostic antigen of 43 kDa always predominated in all CFA preparations . The present method is thus very useful for the rapid production of an antigenic extract which can be readily characterized and used in the serodiagnosis of paracoccidioidomycosis.

Am J Trop Med Hyg, 1991 Jan, 44(1), 83 - 92
Computed tomography and ultrasonography of the adrenal glands in paracoccidioidomycosis . Comparison with cortisol and aldosterone responses to ACTH stimulation; Tendrich M et al.; Fifteen patients with proven disseminated paracoccidioidomycosis (PCM) had computed tomography (CT) and ultrasonography (US) performed to evaluate the form, shape, density and size of their adrenal glands . Plasma and urinary cortisol were determined and adrenal reserve assessed by measuring the cortisol and aldosterone responses to synthetic ACTH . The adrenal CT showed unilateral lesions in two cases and bilateral in another four . The US study showed more frequent alterations, unilateral in seven and bilateral in three subjects . Combining both methods increased the sensitivity to 85% of the cases . All patients had normal plasma cortisol concentrations and normal or increased urinary cortisol excretion . Plasma aldosterone concentration was also normal except in one patient with hypokalemia . Seven patients showed diminished cortisol responses, five had subnormal aldosterone responses and in five plasma aldosterone concentration increased more than normally after stimulation by ACTH . There was an incidence of limited adrenal reserve in 53% of the patients on ACTH stimulation . No correlation was evident between the disorders in adrenal steroid responses to ACTH and changes in morphology revealed by CT and/or US.

Rev Inst Med Trop Sao Paulo, 1991 Jan-Feb, 33(1), 37 - 43
In vitro action of some disinfectants on Paracoccidioides brasiliensis yeast forms; Shikanai-Yasuda MA et al.; The fungicidal action of sodium hypochlorite (0.3, 1, 2.5, 5 and 10%); formaldehyde (2, 5, and 10%); and ethyl alcohol (70%) on yeast forms of Paracoccidioides brasiliensis Pb 18 and a newly-isolated Goiana strain was described . Contact between the fungus and the disinfectants was maintained for 1, 2, 24, 48 and 72 hours at room temperature . Viability was evaluated by the fluorescein diacetate-ethidium bromide treatment, culture in solid and liquid media (36 degrees C and 26 degrees C); yeast to mycelial germination at room temperature; and radiometric study of metabolic activity . All concentrations of disinfectants were found to be effective in inactivating Pb 18 and Goiana strains, except for the 1-hour contact with 2% formaldehyde, in which fluorescein diacetate-ethidium bromide treatment was found to reveal 40 and 27% of viable cells, respectively . The yeast to mycelial germination method was considered to reveal faster and similar results as compared to culture in solid and liquid media.

J Med Vet Mycol, 1991, 29(6), 373 - 80
Studies of natural killer cells in patients with paracoccidioidomycosis; Peracoli MT et al.; The number and activity of natural killer (NK) cells were studied in 34 untreated patients with paracoccidioidomycosis, 20 with the chronic form of the disease and 14 with the acute form . NK cells were detected with monoclonal antibody Leu-11c and the cytotoxic activity was measured using a single cell assay against K562 target cells . Both groups of patients had an increased number of circulating NK cells, their cytotoxic activity being significantly lower than in the healthy controls . These findings may be of importance in the immunological disturbances associated with paracoccidioidomycosis since NK cells exert important immune effector functions and may play a role in resistance against Paracoccidiodes brasiliensis.

Complement Inflamm, 1991, 8(5-6), 288 - 93
Association of major histocompatibility complex class III complement components C2, BF, and C4 with Brazilian paracoccidioidomycosis; de Messias IJ et al.; A genetic influence of the major histocompatibility complex (MHC) on the susceptibility and the development of the different clinical forms of paracoccidioidomycosis (PCM) has been postulated . In the present investigation allotypes of MHC-coded class III gene products (complement components C2, BF, C4A, and B) were determined in 69 Brazilian PCM patients and 225 healthy control individuals matched for ethnic and geographic origin . The frequency of the non-expressed C4B allele (C4B*Q0) was significantly elevated in comparison to the controls (p less than 0.01; Fisher's exact test) . Three out of 69 patients had a complete C4B deficiency as against 2 among 223 control individuals . The C4A*Q0 allele was also more frequent in the patients . Other C4 alleles were not seen to differ between the two groups . The analysis of BF allotypes showed a non-significant predominance of the rarer allele BF*S07 in the patients, whereas no difference in the distribution of C2 alleles was seen . The data on MHC class III association may support the hypothesis of immune response modulation in PCM and suggest a functional genetic role of complement action against the fungus and in the outcome of PCM infection . We conclude that MHC class III products, especially C4B*Q0, are associated with chronic uni- or multifocal PCM and may influence the course of the infection.

Braz J Med Biol Res, 1991, 24(9), 901 - 7
Experimental paracoccidioidomycosis in ethyl alcohol-fed rats; Goldani LZ et al.; 1 . The course of experimental paracoccidioidomycosis was studied in alcohol-fed rats and compared with pair-fed and ad libitum controls . The alcohol-fed rats were fed a regular commercial solid diet plus a 32% (v/v) ethyl alcohol-25% sucrose (w/v) solution 4 weeks prior to the infection and throughout the study . The inoculum was standardized in terms of virulence, age of culture, percentage of multicellular fungal units containing 1-3 cells and viability . 2 . The extent of infection was measured by quantitative counts of fungal units (FU), colony-forming units (CFU) of Paracoccidioides brasiliensis from infected pulmonary tissue and histopathological examination of the main organs involved . The specific humoral response and cell-mediated immunity of the animals were assessed by measuring the specific serum antibody levels by counterimmunoelectrophoresis (CIE) and by the footpad swelling test (FST), respectively . 3 . Alcohol-fed rats and pair-fed controls did not differ significantly in mean body weight or the extent of disease after 5 and 10 weeks of infection . In contrast, the alcohol-fed group when compared with the ad libitum group presented a significantly lower mean body weight (301.6 +/- 0.6 vs 389.6 +/- 0.7 g) and a larger number of fungi recovered from the lung: 81.1 +/- 4.1 x 10(4) vs 13.4 +/- 8.0 x 10(4) FU; 24.3 +/- 8.6 x 10(4) vs 2.9 +/- 0.5 x 10(4) CFU after 5 weeks of infection . There were no significant differences in the specific antibody levels, FST indices, histopathological pattern of frequency of disseminated lesions between groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Braz J Med Biol Res, 1991, 24(8), 807 - 13
Serologic characterization of Paracoccidioides brasiliensis E2 antigen; Siqueira AM et al.; 1 . The behavior of the specific E2 antigen of Paracoccidioides brasiliensis was studied by agarose gel counterimmunoelectrophoresis . When the gel was read immediately after the electrophoretic run no precipitation band was visible . Visualization of the complex was possible only after incubation of the gel at room temperature overnight . 2 . At alkaline pH, the E2 antigen migrates in the direction of the cathode, as do the immunoglobulins . The higher sensitivity of counterimmunoelectrophoresis when compared to double immunodiffusion for the diagnosis of paracoccidioidomycosis is due to the presence of antibodies directed against antigens which migrate to the anode . 3 . The use of specific antiserum to E2 antigen as reference permits the double immunodiffusion method to be a very sensitive test for the specific serodiagnosis of paracoccidioidomycosis.

J Med Vet Mycol, 1991, 29(4), 243 - 53
Antigens of diagnostic value in three isolates of Paracoccidioides brasiliensis; Casotto M et al.; Yeast cellular extracts of three isolates of Paracoccidioides brasiliensis were tested by the Western-blot technique against 53 sera from patients with paracoccidioidomycosis (PCM) . Numerous antigens were recognized by the sera but only five were specific for (58, 57, 48, 44 and 23 kDa) . These specific antigens had the same relative molecular mass in the different isolates but their reactivity with sera from PCM patients was variable . An antigen in isolate 688, which was specific for PCM, had an apparent molecular mass of 48 kDa . Although a 48 kDa antigen was also present in isolates B339 and 1789.88 it was not specific for PCM, demonstrating antigenic variability among isolates . The 44 kDa antigen in isolate B339 and the 44 and 48 kDa antigens in isolate 688 reacted with a rabbit antiserum raised against a 43 kDa glycoprotein, a specific antigen used in the diagnosis of PCM . There was no correlation between the specific antigen(s) detected and the clinical form of the disease.

Cell Immunol, 1991 Jan, 132(1), 236 - 45
Kinetics and requirements for activation of macrophages for fungicidal activity: effect of protein synthesis inhibitors and immunosuppressants on activation and fungicidal mechanism; Brummer E et al.; Peritoneal-and pulmonary macrophages can be activated in vitro with lymphokines (LK) or IFN-gamma, without exogenous lipopolysaccharide, for fungicidal activity against several pathogenic fungi . However, neither the biochemical nor metabolic events of the activation process or of the effector phase have been defined . In the present work we sought to elucidate these events with time-course studies using inhibitors of protein synthesis as well as immunosuppressive agents . We found that protein synthesis inhibitors abrogated the activation process, because cycloheximide (CHX) (1-2 micrograms/ml) prevented activation of macrophages for fungicidal activity against Candida albicans, Blastomyces dermatitidis, and Paracoccidioides brasiliensis . Blocking of the activation process by CHX was not due to macrophage cytotoxicity, and CHX did not impair the ability of nonactivated macrophages to kill Candida parapsilosis . In kinetic studies we showed that activation of macrophages was induced in 4 hr of LK treatment and that CHX had no effect if added after this time . In contrast to CHX, therapeutic concentrations of hydrocortisone (HC), such as less than or equal to 5 micrograms/ml, or cyclosporin A (CsA), 5 micrograms/ml, did not significantly inhibit LK activation of macrophages for killing of fungi . In the effector phase, the fungicidal capacity of activated macrophages in short-term (less than or equal to 4 hr) killing assays could not be abrogated by CHX (5 micrograms/ml), HC (100 micrograms/ml), or CsA (10 micrograms/ml) . These results demonstrate that the activation but not the effector mechanism of macrophages for fungicidal activity is blocked by inhibition of protein synthesis . In contrast, therapeutic concentrations of HC or CsA may not interfere with activation of macrophages or their killing mechanisms, thus providing a rationale for antifungal immunotherapy in certain clinical situations (e.g., infection in the immunosuppressed patient).

Microbiol Immunol, 1991, 35(3), 167 - 74
Lymphocyte subpopulations and cytokine production in paracoccidioidomycosis patients; Bava AJ et al.; Despite the postulated role of the immune system in the control of the infection by Paracoccidioides brasiliensis, only a few studies have addressed this point in patients . The determination of total lymphocytes and their subpopulations in 6 untreated patients with the chronic form of paracoccidiodomycosis showed that half of them were lymphopenic, because of low number of CD4+ T-lymphocytes . All patients had low CD4/CD8 ratios . On the contrary, B-lymphocytes were normal in all patients . An additional patient, studied on treatment with ketoconazole, had normal lymphocyte counts in all subpopulations, as did one of the patients previously studied at diagnosis when he received specific antimycotic treatment . The production of interferon and tumor necrosis factor, determined by bioassay in supernatants of mononuclear blood cells of the patients, induced by interleukin 2 in vitro was significantly lower than that of normal subjects . These results show that patients with paracoccidioidomycosis have a defect in blood lymphocyte subsets as well as in the ability to produce regulatory cytokines.

Rev Soc Bras Med Trop, 1990 Oct-Dec, 23(4), 213 - 5
{Epidemiologic survey with histoplasmin and paracoccidioidine in Arapiraca-Alagoas}; Santos MC et al.; Paracoccidioidin and histoplasmin intradermic tests were performed in 107 inhabitants in Arapiraca, Alagoas . The epidemiological survey was performed in both sexes and the ages varied from 2 to 71 years . Positive results with paracoccidioidin test were recorded in 11% and the histoplasmin test was positive in 14% . With the positivity of paracoccidioidin in the histoplasmin positive population the result shows a percentage of 33.33%, while with the positivity of histoplasmin in the paracoccidioidin positive population was 41%.

Trans R Soc Trop Med Hyg, 1990 Sep-Oct, 84(5), 734 - 7
Preparation of murine monoclonal antibodies against the yeast phase of the dimorphic fungus Sporothrix schenckii; Hamilton AJ et al.; Three murine monoclonal antibodies (Mabs) were raised against a cytoplasmic antigen of the yeast phase of the pathogenic fungus Sporothrix schenckii using a modification of standard hybridoma technology incorporating the immunosuppressive drug cyclophosphamide . When tested for species-specificity within the pathogenic dimorphic fungi one of these Mabs (S5) showed little cross-reactivity by enzyme-linked immunosorbent assay and Western blot, though there was some recognition of Paracoccidioides brasiliensis antigen . This Mab recognized a 70-75 kDa molecule on reduced Western blots of S . schenckii antigen . The other two Mabs (S12 and S15) showed cross-reactivity with all dimorphic fungal antigens tested, though they appeared to recognize a molecule of similar molecular weight . This is the first report of any attempt to raise species-specific Mabs against this important causative agent of dermatological disease.

Mycopathologia, 1990 Sep, 111(3), 173 - 80
Alterations in the pathogenicity of one Paracoccidioides brasiliensis isolate do not correlative with its in vitro growth; Kashino SS et al.; The in vitro subcultivation of some microorganisms for long periods causes measurable loss of their pathogenicity, which can be reverted by reisolation from infected hosts . We compared the pathogenicity and the in vitro growth pattern of one P . brasiliensis isolate (Pb 18) in its yeast phase, using the following samples: 1) The original pathogenic Pb 18 (OP) . 2) Pb 18 attenuated by continuous in vitro subcultivation (AT) . 3) Pb 18 (AT) reisolated from susceptible B10.A mice (RS) . 4) Pb 18 (AT) reisolated from resistant A/SN mice (RR) . Pathogenicity was evaluated by anatomopathology and mortality of mice infected i.p . with 5 x 10(6) fungi . Median survival times of mice infected with OP ranged from 74 to 117 days during the first 51 months of subculturing; with more cycles of subculturing the median survival time increased, reaching 250 days at the 64th month . This indicated decreasing virulence of OP during this period of subculturing . Survival of mice infected with RS and RR was respectively 112 and 123 days, which is similar to the behavior of the OP variant . The in vitro growth curve profile of RR showed significantly higher numbers of total and viable yeasts than the other studied variant . These results show that: 1) Pb 18 isolate loses its pathogenicity by continuous subcultivation . This phenomenon is reverted by reisolation from mice, independently from their susceptibility to the fungus; 2) the in vitro growth patterns of Pb 18 do not correlate with alterations in pathogenicity but are influenced by the host's environment.

Cutis, 1990 Sep, 46(3), 227 - 34
Lobomycosis (keloidal blastomycosis): case reports and overview; Fuchs J et al.; Lobomycosis is a deep fungal disease of the skin without involvement of internal organs or mucous membranes . The disease is characterized by skin nodules and plaques resembling keloid involving the earlobes, distal parts of the upper and lower extremities, and buttocks . In severe cases, large skin areas can be covered by disseminated or grouped and confluent nodules . Most cases are reported from South and Central America . The fungus Paracoccidioides (Glenosporella) loboi is abundant in lesions but is extremely difficult to culture . Lobomycosis is resistant to chemotherapy, but in some cases it can successfully be treated by excision . Although the diagnosis is easily established by its typical clinical, histologic, and microbiological features, it is often misdiagnosed by physicians not familiar with the disease . We describe here five patients and present an overview of this rare disease.

Mycopathologia, 1990 Sep, 111(3), 169 - 72
Adenosine triphosphatase in yeast phase Paracoccidioides brasiliensis; Campo-Aasen I et al.; We have previously detected various enzymatic activities in P . brasiliensis . In the present study we have examined Adenosine Triphosphatase (ATPase) activity in yeast phase cultures of P . brasiliensis of increasing age . We employed Wachstein and Meisel's method and electron microscopy and found specific electron-dense deposits indicating ATPase activity to be present in the cytoplasm around vacuoles . Their distribution varied according to age . Deposits decreased or became absent in old cultures . We assume that in P . brasiliensis, ATPase is involved (as in other systems) with transport of Na+ and K+.

Rev Inst Med Trop Sao Paulo, 1990 Sep-Oct, 32(5), 370 - 4
{Obtention of Histoplasma capsulatum exoantigens in neopeptone, glucose, thiamine and asparagine (NGTA) media}; Garcia NM et al.; The purpose of this work is obtaining exocellular antigens H and M from 4 H . capsulatum strains using NGTA medium (neopeptone, glucose, thiamine and asparagine) for periods of 1, 2 and 3 months, at 36 degrees C and continuously shaken . The exocellular antigens were evaluated by double immunodiffusion test against H . capsulatum rabbit antiserum, 7 histoplasmosis sera, 4 paracoccidioidomycosis sera and a reference antigen and antibody furnished by C.D.C . (Atlanta--USA) . Except for the exocellular antigen from strain B.679 with 1 month of culture, all exocellular antigens obtained from the strains B.679, 58 and O187 showed the H and M bands . The A.811 strain demonstrated only the fraction H . All the exocellular antigens reacted positively with sera from histoplasmosis patients, except those obtained from strains 58 and B.679 with 1 month of culture . With regard to paracoccidioidomycosis patients sera, the exocellular antigens from strains 58 and O187 did not cross-react with them.

J Clin Microbiol, 1990 Aug, 28(8), 1766 - 9
Preparation of species-specific murine monoclonal antibodies against the yeast phase of Paracoccidioides brasiliensis; Figueroa JI et al.; A panel of four murine monoclonal antibodies showing species specificity for the yeast phase of the pathogenic dimorphic fungus Paracoccidioides brasiliensis was produced by using a modification of the standard monoclonal antibody technology . This involved the use of the immunosuppressive drug cyclophosphamide to suppress the immune response of test animals to fungi showing cross-reactivity, i.e., to Histoplasma capsulatum . One monoclonal antibody, P4, which had a high titer by enzyme-linked immunosorbent assay, was shown to recognize a linear antigenic epitope of P . brasiliensis at a molecular size of 70,000 to 75,000 daltons by Western blot (immunoblot) analysis . The potential use of these monoclonal antibodies, which are the first species-specific probes to P . brasiliensis that have been produced, in the field of serodiagnosis is discussed.

Am J Trop Med Hyg, 1990 Aug, 43(2), 200 - 6
Antibody response to the 43 kDa glycoprotein of Paracoccidioides brasiliensis as a marker for the evaluation of patients under treatment; Giannini MJ et al.; Sera of patients with paracoccidioidomycosis contained IgG-, IgA-, and IgM-specific antibodies to a 43 kDa antigen contained in the filtrate of a culture of Paracoccidioides brasiliensis . IgG- and IgA-specific antibodies were present in all observed patients . The IgM response was more frequent in acute cases, and the mean titers of IgG- and IgM-specific antibodies were higher in the acute forms . By the fourth month of chemotherapy, there was a decay of IgG, IgA, and IgM antibody titers to this antigen in acute cases, correlating with clinical improvement . The detection of IgG and IgA antibodies and the sequential determination of antibodies to the 43 kDa glycoprotein may be useful tools for serodiagnosis and evaluation of therapeutic efficacy.

Infect Immun, 1990 Aug, 58(8), 2628 - 36
Ultrastructure of phagocytosed Paracoccidioides brasiliensis in nonactivated or activated macrophages; Brummer E et al.; Transmission electron microscopy was used to study ultrastructures in Paracoccidioides brasiliensis yeast cells after ingestion by nonactivated or cytokine-activated murine peritoneal macrophages . Yeast cells ingested by nonactivated macrophages had typical bi- and trilayered cell walls, plasma membranes, mitochondria, nuclei, vacuoles, etc., which remained intact for 24 h of coculture . In contrast, yeast cells ingested by activated macrophages exhibited abnormal mitochondrial ultrastructures within 4 h of interaction . Subsequent events that occurred were the formation of several clear vacuoles per cell, disintegration of the cytoplasm, and development of empty cells with intact walls . These findings provide, for the first time, insights into stepwise damage to fungal cells by activated macrophages (of particular interest in this instance because of prior evidence that the damage is due to nonoxidative mechanisms) and give possible clues regarding fungicidal mechanisms.

J Biol Chem, 1990 Jul 5, 265(19), 11185 - 92
The sequence of the cyo operon indicates substantial structural similarities between the cytochrome o ubiquinol oxidase of Escherichia coli and the aa3-type family of cytochrome c oxidases; Chepuri V et al.; The cytochrome o complex is one of two ubiquinol oxidases in the aerobic respiratory system of Escherichia coli . This enzyme catalyzes the two-electron oxidation of ubiquinol-8 which is located in the cytoplasmic membrane, and the four-electron reduction of molecular oxygen to water . The purified oxidase contains at least four subunits by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and has been shown to couple electron flux to the generation of a proton motive force across the membrane . In this paper, the DNA sequence of the cyo operon, containing the structural genes for the oxidase, is reported . This operon is shown to encode five open reading frames, cyoABCDE . The gene products of three of these, cyoA, cyoB, and cyoC, are clearly related to subunits II, I, and III, respectively, of the eukaryotic and prokaryotic aa3-type cytochrome c oxidases . This family of cytochrome c oxidases contain heme a and copper as prosthetic groups, whereas the E . coli enzyme contains heme b (protoheme IX) and copper . The most striking sequence similarities relate the large subunits (I) of both the E . coli quinol oxidase and the cytochrome c oxidases . It is likely that the sequence similarities reflect a common molecular architecture of the two heme binding sites and of a copper binding site in these enzymes . In addition, the cyoE open reading frame is closely related to a gene denoted ORF1 from Paracoccus dentrificans which is located in between the genes encoding subunits II and III of the cytochrome c oxidase of this organism . The function of the ORF1 gene product is not known . These sequence relationships define a superfamily of membrane-bound respiratory oxidases which share structural features but which have different functions . The E . coli cytochrome o complex oxidizes ubiquinol but has no ability to catalyze the oxidation of reduced cytochrome c . Nevertheless, it is clear that the E . coli oxidase and the aa3-type cytochrome c oxidases must have very similar structures, at least in the vicinity of the catalytic centers, and they are very likely to have similar mechanisms for bioenergetic coupling (proton pumping).

J Biol Chem, 1990 Jun 15, 265(17), 9793 - 9
Complete amino acid sequence of the cytochrome subunit and amino-terminal sequence of the flavin subunit of flavocytochrome c (sulfide dehydrogenase) from Chlorobium thiosulfatophilum; Van Beeumen J et al.; The complete amino acid sequence of the 86-residue heme subunit of flavocytochrome c (sulfide dehydrogenase) from the green phototrophic bacterium Chlorobium thiosulfatophilum strain Tassajara has been determined as follows: APEQSKSIPRGEILSLSCAGCHGTDGKSESIIPTIYGRSAEYIESALLDFKSGA- RPSTVMGRHAKGYSDEEIHQIAEYFGSLSTMNN . The subunit has a single heme-binding site near the N terminus, consisting of a pair of cysteine residues at positions 18 and 21 . The out-of-plane ligands are apparently contributed by histidine 22 and methionine 60 . The molecular weight including heme is 10,014 . The heme subunit is apparently homologous to small cytochromes c by virtue of the location of the heme-binding site and its extraplanar ligands . However, the amino acid sequence is closer to Paracoccus sp . cytochrome c554(548) (37%) than it is to the heme subunit from Pseudomonas putida p-cresol methylhydroxylase flavocytochrome c (20%) . The flavocytochrome c heme subunit is only 14% similar to the small cytochrome c555 also found in Chlorobium . Secondary structure predictions suggest N- and C-terminal helices as expected, but the midsection of the protein probably folds somewhat differently from the small cytochromes of known three-dimensional structure such as Pseudomonas cytochrome c551 . Analyses of the residues near the exposed heme edges of the cytochrome subunits of P . putida and C . thiosulfatophilum flavocytochromes c (assuming homology to proteins of known structure) indicate that charged residues are not conserved, suggesting that electrostatic interactions are not involved in the association of the heme and flavin subunits . The N-terminal sequence of the flavoprotein subunit of flavocytochrome has also been determined . It shows no similarity to the comparable region of the p-cresol methylhydroxylase flavoprotein subunit from P . putida . The flavin-binding hexapeptide, isolated and sequenced earlier (Kenney, W . C., McIntire, W., and Yamanaka, T . (1977) Biochim . Biophys . Acta 483, 467-474), is situated at positions 40-46.

J Clin Microbiol, 1990 Jun, 28(6), 1188 - 93
Characterization of the cellular antigens of Paracoccidioides brasiliensis yeast form; Casotto M; Antigenic components of the yeast extract of Paracoccidioides brasiliensis Linder 2511 cultured for 3, 8, 20, 30, and 60 days were examined by the Western blot (immunoblot) technique . The 3-day extract was chosen for characterization of the antigenic components because its stability did not vary with time and it contained all antigens identified by patient sera . Antibodies to cross-reacting antigens of P . brasiliensis extracts were detected in sera from patients with histoplasmosis, candidiasis, and aspergillosis . The 58-, 57-, 21-, and 16-kilodalton (kDa) antigens were specific for P . brasiliensis, while the 48- and 45-kDa antigens were specific for paracoccidioidomycosis . The Western blot technique is a useful tool for the diagnosis of disease and revealed heterogeneity in the responses of patient sera . The combination of the 58-, 57-, and 45-kDa proteins confirmed a diagnosis of paracoccidioidomycosis (87% of the cases).

Rev Cubana Med Trop, 1990 May-Aug, 42(2), 272 - 8
{The use of exoantigens for identifying Histoplasma capsulatum}; Fernandez Andreu C et al.; The exoantigen test has been successfully used in the last years for the immunological identification of pathogenic fungi . Twenty seven straing presumptively identified as Histoplasma capsulatum by morphologic methods, were studied by such test . The exoantigens obtained from each strain were studied by immunoprecipitation technique in front of histoplasmosis control sera experimentally obtained in rabbits . Reference strains of Histoplasma capsulatum . Histoplasma duboisii and Paracoccidioides brasiliensis were used . Of the strains studied, 96% was identified as H . capsulatum by the exoantigen test, a simple, rapid and highly specific technique greatly useful in the laboratories of Microbiology.

Mycoses, 1990 Mar, 33(3), 111 - 5
Influence of serum on in vitro digestion of Paracoccidioides brasiliensis by neutrophils; Goihman-Yahr M et al.; Serum from patients with paracoccidioidomycosis (PARA) did not block digestive abilities of neutrophils (PMNs) from healthy individuals against Paracoccidioides brasiliensis . Conversely, serum from healthy donors did not enhance digestive capacities of PMNs from patients with PARA vis a vis the causative organism . We conclude that the specific digestive defect present in PMNs from patients with PARA is not mediated by serum factors.

Rev Inst Med Trop Sao Paulo, 1990 Mar-Apr, 32(2), 116 - 20
{Histoplasmin and paracoccidioidin reactions in Serra de Pereiro . (Ceara State--Brazil)}; Diogenes MJ et al.; Intradermal tests using histoplasmin and paracoccidioidin antigens were performed in 138 persons from Pereiro, Ceara, Brazil . The results were positive in 61.5% and 32.5% with histoplasmin and paracoccidioidin antigens respectively . These results suggest infection by H . capsulatum and P . brasiliensis in the people living in the studied area . New studies are necessary to detect histoplasmosis and paracoccidioidomycosis as clinical diseases in that region.

J Gen Microbiol, 1990 Feb, 136 ( Pt 2), 331 - 5
A murine monoclonal antibody exhibiting high species specificity for Histoplasma capsulatum var . capsulatum; Hamilton AJ et al.; A monoclonal antibody (mAb) exhibiting a high degree of species specificity for the yeast phase of the dimorphic fungus Histoplasma capsulatum was produced by a modification of the standard mAb production protocol . The technique for generating mAbs involved the use of the immunosuppressive drug cyclophosphamide to diminish the response in mice to immunodominant cross-reactive epitopes . This mAb exhibited clear specificity and did not react by ELISA with the closely related genera Blastomyces, Paracoccidioides and Sporothrix . In Western blots it recognized a linear determinant on a 70-75 kDa molecule in H . capsulatum antigen, with an extremely faint reactivity to antigens of identical molecular mass derived from Sporothrix and Paracoccidioides, and no reactivity against Blastomyces antigen.

J Med Vet Mycol, 1990, 28(3), 253 - 6
Isolation and characterization of a Paracoccidioides brasiliensis strain from a dogfood probably contaminated with soil in Uberlândia, Brazil; Ferreira MS et al.; We report the isolation of a strain of Paracoccidioides brasiliensis from a dogfood, probably contaminated with soil, in a Brazilian city . The fungus was isolated on appropriate culture media, and when inoculated into a guinea-pig testis produced orchitis with abundant fungal elements . Histopathology of sections of the testicle showed an inflammatory reaction with P . brasiliensis inside monocytes . Immunological identification confirmed the identity of the isolate.

Mem Inst Oswaldo Cruz, 1989 Jan-Mar, 84(1), 53 - 60
{Detection of amastigotes in cutaneous and mucocutaneous leishmaniasis using the immunoperoxidase method, using polyclonal antibody: sensibility and specificity compared with conventional methods of diagnosis}; Salinas G et al.; The indirect immunoperoxidase method was evaluated in 265 biopsies with the purpose of increasing the sensitivity of the diagnostic histopathology of tegumentary lesions caused by subspecies of the Leishmania braziliensis complex . A diagnosis of leishmaniasis was established by parasitological methods (181) or clinical criteria (12) in 193 patients (72.8%) . In the latter group of confirmed cases standard histochemistry and immunoperoxidase were compared with direct examination of tissue scraping and culture of lesion aspirates . The detection and localization of amastigotes was more efficient using the immunoperoxidase method (61.3%) than conventional histopathology with hematoxilin and eosin (34.6%) or direct examination of tissue scraping (43.9%) . However, culture of lesion aspirates was the most sensitive procedure (89.8%) . The efficiency of the immunoperoxidase method was greater in recent lesions, being positive in 75% of cases with less than 3 months evolution, while 55.6%, 37.5%, and 21.1% of cases with lesion evolution of 3-5.9, 6-11, and 12 months or greater, respectively, were positive . The combined use of the direct examination of lesion scraping and immunoperoxidase applied to histological sections of the biopsy from the lesion border allowed an etiologic diagnosis of 72% of confirmed cases . Cross-reactivity was observed with Paracoccidioides braziliensis but not with Mycobacterium leprae, Sporothrix schenckii, or Histoplasma capsulatum.

J Med Vet Mycol, 1990, 28(1), 67 - 76
Treatment of paracoccidioidomycosis with itraconazole; Naranjo MS et al.; Forty-seven patients with active paracoccidioidomycosis were treated with itraconazole . The majority of the patients (45) had the chronic adult form of the disease while the remaining exhibited the juvenile form . All of the patients received itraconazole, 100 mg day-1, with the exception of two for whom the clinical condition necessitated a larger dose . The mean duration of therapy was 6 months, with a range of 3-24 months . Clinically, the skin and mucous membrane lesions as well as abdominal pain, auscultatory alterations and dysphonia improved markedly during treatment with a concomitant increase in weight . However, other symptoms (cough, expectoration and dyspnoea) persisted as sequelae in some cases . As expected adrenal insufficiency also persisted . The radiological lesions showed a gradual decrease of both scattered and confluent infiltrates, present in 67% of patients before, and in 13% at the end of treatment . On the other hand, fibrosis became more evident at the termination of therapy, increasing from 62% of patients at the beginning of therapy to 81% at the end . The mycological tests (direct examination and cultures) became negative during the first month of treatment in 42 patients (87%) . A decline in specific antibody titres was observed in 72% of patients by the end of treatment . Evaluation of therapy by means of a scoring system indicated complete resolution of the disease in one patient (2%), marked improvement in 42 (89%) and minor improvement in four (8.5%); none of the patients showed a deterioration of their disease during therapy . Of fifteen patients who were followed up for 12 months post-therapy none showed clinical relapse during this period . The results of this study confirm the efficacy of itraconazole in the treatment of paracoccidioidomycosis.

Mycopathologia, 1990 Jan, 109(1), 13 - 7
Virulence of Paracoccidiodes brasiliensis: the influence of in vitro passage and storage; Brummer E et al.; Stability of virulence in P . brasiliensis isolates was studied with respect to the in vitro culture history and methods used for storage . Virulence in yeast-form P . brasiliensis isolates was tested in a chronic pulmonary murine model of paracoccidiodomycosis where progression of disease was quantitated in terms of colony forming units recoverable from lungs . Four isolates of P . brasiliensis, including recently isolated form patients or experimental animals, caused chronic progressive disease . Two isolates with a history of subculturing showed attenuation by causing resolving but chronic disease . An attenuated isolate became avirulent subsequent to 15 more years of subculturing . These findings suggest that virulence of P . brasiliensis can be attenuated or lost subsequent to cycles of subculturing over long periods . Our data suggest that the use of fresh P . brasiliensis isolates may be needed to provide reproducible virulence for experimental systems.

Braz J Med Biol Res, 1990, 23(2), 151 - 61
Reticulin antibodies: relationship with endocardium-vessel-interstitium antibodies and heterophilic antibodies; Viero RM et al.; 1 . Indirect immunofluorescence was used to compare reticulin antibodies, endocardium-vessel-interstitium (EVI) antibodies and heterophilic antibodies in serum samples from patients with degenerative and inflammatory diseases, Chagas' disease, or paracoccidioidomycosis and from healthy blood donors . 2 . The antigen substrates used were rat, mouse and human liver for reticulin antibodies, mouse and human heart and skeletal muscle for EVI factor, and rat heart and rat, mouse and human kidney and stomach for heterophilic antibodies . 3 . The three antibodies produced extremely similar fluorescent patterns and were present simultaneously in a large proportion of reticulin-positive serum samples . 4 . The three antibodies were significantly absorbed by heterophilic antigens such as sheep and rat red blood cells and guinea pig kidney, by Trypanosoma cruzi (an antigen associated with EVI factor) and by Paracoccidioides brasiliensis . However, they did not react with several human tissues . 5 . These results suggest that reticulin and EVI antibodies can be considered to be heterophilic antibodies.

J Med Vet Mycol, 1990, 28(2), 151 - 7
Study of antibodies in paracoccidioidomycosis: follow-up of patients during and after treatment; Ferreira-da-Cruz MF et al.; The relevance of the humoral response in the prognosis of paracoccidioidomycosis was assessed by measuring the serological responses of individual patients to Paracoccidioides brasiliensis by double immunodiffusion (DID) . Sixty-six patients with paracoccidiodomycosis were studied . Sera from 31 individuals were tested before and during treatment with sulfonamide (Group I) . Sera from a further 35 individuals were tested after completion of a 2-year course of treatment (Group II) . In Group I, clinical improvement was associated with a decrease in antibody titer in all patients . The only patient in this group who had a clinical relapse during specific treatment presented with a 4-fold increase in antibody titer immediately before relapse . In Group II, nine patients remained antibody positive at follow-up (61.9 +/- 40.0 months), despite their good physical health, indicating that the detection of antibodies to P . brasiliensis by the DID test does not necessarily indicate active disease . These data suggest that changes in antibody titers to P . brasiliensis detected by DID may be useful indicators of the extent of active disease . Measurement of antibody titers may be valuable for determining the prognosis of the infection and for deciding on a suitable treatment protocol.

J Med Vet Mycol, 1990, 28(1), 35 - 46
Transfer of cell-mediated immunity to Paracoccidioides brasiliensis in hamsters with dialysable leukocyte extracts; Peracoli MT et al.; Dialysable leukocyte extracts (DLE) were obtained from lymph nodes and spleen cells of hamsters immunized with Paracoccidioides brasiliensis (immune DLE) or from non-immunized hamsters (non-immune DLE) . Sensitivity to P . brasiliensis antigen (PbAg) was transferred by immune DLE to normal recipient hamsters and could be detected by the macrophage migration inhibition (MIF) assay on day three and by a positive skin test to PbAg on day seven after DLE inoculation . This reactivity persisted for 120 days . The specificity of DLE was studied by inoculation of immune or non-immune DLE into normal hamsters and transfer was evaluated 7 days later by the MIF assay and by skin tests to PbAg, Candida albicans antigen (CaAg), sonically treated bacille Calmette-Guerin (BCG) (SBCG) and ovalbumin (OVA) . The immune DLE recipients showed a reactivity to PbAg that was significantly stronger than that of the nonimmune DLE recipients although both groups of recipient animals showed reactivity to CaAg and SBCG . The results suggest that both DLE preparations contain a non-specific antigen augmenting factor . This factor may stimulate the minimal background response to the antigens in nonimmunized recipient hamsters.

Mycoses, 1989 Dec, 32(12), 603 - 8
Functions of polymorphonuclear leukocytes and individuality of Jorge Lobo's disease: absence of the specific leukocyte digestive defect against Paracoccidioides brasiliensis; Goihman-Yahr M et al.; Peripheral blood neutrophils (PMNs) from a patient with Jorge Lobo's disease (JLD) digested well phagocytosed Paracoccidioides brasiliensis . We found no circulating antibodies against P . brasiliensis in the patient's serum . Such neutrophils showed myeloperoxidase activity and also digested normally phagocytosed Candida albicans . We had previously reported the presence of a specific digestive deficiency of PMNs from patients with paracoccidioidomycosis (PARA) vis a vis P . brasiliensis . Current findings provide new information about leukocyte functions in JLD and bolster the view that JLD, PARA and their respective causative microorganisms are distinct.

J Clin Microbiol, 1989 Dec, 27(12), 2842 - 5
Detection of the 43,000-molecular-weight glycoprotein in sera of patients with paracoccidioidomycosis; Mendes-Giannini MJ et al.; The 43,000-molecular-weight (43K) soluble glycoprotein was detected in sera of patients with paracoccidioidomycosis by the immunoblot technique by using as the probe rabbit monospecific antisera to this fraction . The 43K antigen was present before treatment in sera of patients with the acute (juvenile) form; it started to disappear from circulation after 10 months of chemotherapy, and it was undetectable after 2 years of treatment . In the chronic cases, the 43K antigen was detected in patients without treatment, and it was absent in the healed cases . The detection of the 43K protein specific to Paracoccidioides brasiliensis may be important for its diagnostic value as well as for modulation of the host immune response.

Mycopathologia, 1989 Nov, 108(2), 89 - 93
Polyclonal B cell activation in paracoccidioidomycosis . Polyclonal activation in paracoccidioidomycosis; Chequer-Bou-Habib D et al.; Paracoccidioidomycosis (PCM) is often associated with hypergammaglobulinemia and increased serum levels of circulating immune complexes (CIC) . In order to investigate whether polyclonal B lymphocyte activation (PBA) is a current process in PCM, we measured the numbers of IgG secreting cells (IgG SC) in the peripheral blood of 16 patients and of 8 healthy controls . The numbers of IgG SC were found to be significantly elevated in PCM patients . We also observed increased serum levels of IgG, IgA and CIC . These data reflect an activation of B lymphocytes in PCM patients.

Mycopathologia, 1989 Nov, 108(2), 107 - 12
Evaluation of acetylator phenotype, renal function and serum sulfadiazine levels in patients with paracoccidioidomycosis treated with cotrimazine (a combination of sulfadiazine and trimethoprim); Barraviera B et al.; The authors evaluated the relationships among renal function, acetylator phenotype and serum sulfadiazine levels in 22 patients with paracoccidioidomycosis treated with 1 tablet of cotrimazine (a combination of 820 mg sulfadiazine and 180 mg trimethoprim) administered orally every 12 hours . Fifteen patients (68.18%) presented free sulfadiazine levels above 50 micrograms/ml, 6(27.28%) presented serum levels above 40 micrograms/ml, and 1(4.54%), levels lower than 40 micrograms/ml, this being the patient in which treatment failed . The highest free sulfadiazine levels were obtained in slow acetylator patients with reduced renal function . One patient with neuroparacoccidioidomycosis presented free sulfadiazine levels in cerebrospinal fluid corresponding to 55% of the serum levels . Finally, the authors consider cotrimazine to be an important therapeutic alternative for neuroparacoccidioidomycosis and conclude that administration every 12 hours can provide therapeutic sulfadiazine levels . They also suggest that when the sulfadiazine-trimethoprim combination is used, the therapeutic levels of sulfadiazine should be above 40 micrograms/ml.

Antimicrob Agents Chemother, 1989 Sep, 33(9), 1641 - 4
Inhibition of growth of the dimorphic fungus Paracoccidioides brasiliensis by ajoene; San-Blas G et al.; Ajoene, a garlic-derived compound that prevents platelet activation, inhibited the growth of Paracoccidioides brasiliensis, a fungal pathogen for humans, by affecting the integrity of the fungal cytoplasmic membrane . This action may be the basis for the study of ajoene as a possible specific antifungal drug.

Rev Prat, 1989 Sep 1, 39(19), 1675 - 82
{Histoplasmosis and other imported mycoses in 1989}; Drouhet E et al.; The most common systemic mycoses imported into France are the histoplasmosis caused by H . capsulatum, which comes from Black Africa and from the French overseas departments and territories (Guyane, Martinique, Guadeloupe, New Caledonia), and the histoplasmosis caused by the larger H . duboisii, which is exclusively Central African . In recent years, histoplasmosis has become an opportunistic infection in immunocompromised patients, particularly those with AIDS . Blastomycosis, imported from North America as well as from North Africa and Central Africa, and paracoccidioidomycosis, imported from Latin America, are pulmonary mycoses with cutaneous manifestations on the face and extremities and with various deep localizations which often follow a chronic course . Coccidioidomycosis, strictly limited to the desertic regions in the western part of the American continent, is also a pulmonary mycosis with multiple granulomas in the skin, bones, lymph nodes and meninges . Penicilliosis caused by Penicillium marneffei is a mycosis from South-East Asia which has recently been observed in European AIDS patients who had travelled in that part of the world.

Rev Med Panama, 1989 Sep, 14(3), 135 - 8
{Prevalence of Paracoccidioides brasiliensis infection in Panamanian children . Preliminary report}; de Martin MC et al.; The prevalence of Paracoccidioides brasiliensis infection in Panamanian children was demonstrated to be 12.73% (14/110) by skin reactivity to 50 micrograms of yeast derived antigen . Boys were infected more frequently than girls - 1.45:1 . The earliest age of infection was 2 years, and a direct relationship was noted between age and frequency of infections (as the children became older, more of them became infected); boys more so than girls . Thirteen of 14 (93%) of the infected juvenile population live in a rural environment.

Infect Immun, 1989 Aug, 57(8), 2289 - 94
Intracellular multiplication of Paracoccidioides brasiliensis in macrophages: killing and restriction of multiplication by activated macrophages; Brummer E et al.; The effect of coculturing yeast-form Paracoccidioides brasiliensis with murine cells was studied . Coculture of resident peritoneal or pulmonary macrophages with P . brasiliensis for 72 h dramatically enhanced fungal multiplication 19.3 +/- 2.4- and 4.7 +/- 0.8-fold, respectively, compared with cocultures with lymph node cells or complete tissue culture medium alone . Support of P . brasiliensis multiplication by resident peritoneal macrophages was macrophage dose dependent . Lysates of macrophages, supernatants from macrophage cultures, or McVeigh-Morton broth, like complete tissue culture medium, did not support multiplication of P . brasiliensis in 72-h cultures . Time course microscopic studies of cocultures in slide wells showed that macrophages ingested P . brasiliensis cells and that the ingested cells multiplied intracellularly . In sharp contrast to resident macrophages, lymphokine-activated peritoneal and pulmonary macrophages not only prevented multiplication but reduced inoculum CFU by 96 and 100%, respectively, in 72 h . Microscopic studies confirmed killing and digestion of P . brasiliensis ingested by activated macrophages in 48 h . These findings indicate that resident macrophages are permissive for intracellular multiplication of P . brasiliensis and that this could be a factor in pathogenicity . By contrast, activated macrophages are fungicidal for P . brasiliensis.

Rev Saude Publica, 1989 Aug, 23(4), 285 - 91
{Anatomo-clinical diagnosis correlation . Retrospective assessment of the clinical diagnosis in necropsies}; de Almeida MC et al.; The post mortem findings in 200 autopsies were compared with the clinical diagnoses . Twenty diseases were analysed with respect to clinico-pathological correlation . In relation to the respective totals the diseases most frequently missed clinically were (false negative) pyelonephritis (100%), pulmonary embolus (87.50%) and bronchopneumonia (58.16%) . In relation to the respective totals the clinical diagnoses less frequently confirmed (false positive) were tuberculosis (69.56%), paracoccidioidomycosis (57.14%), sepsis (53.13%) and Chagas' disease (44.44%) . There was clinicopathological agreement in 97 autopsies (48.50%) . In 19 cases (9.50%) if the diagnostic error had been detected during life this probably would have changes the prognosis . The findings are discussed in the light of previous studies . The importance of routine post-mortem examination and clinico-pathological correlation is stressed.

Biochemistry, 1989 Jul 25, 28(15), 6318 - 22
Redox protein electron-transfer mechanisms: electrostatic interactions as a determinant of reaction site in c-type cytochromes; Cheddar G et al.; The effect of ionic strength on the rate constant for electron transfer has been used to determine the magnitude and charge sign of the net electrostatic potential which exists in close proximity to the sites of electron transfer on various c-type cytochromes . The negatively charged ferricyanide ion preferentially reacts at the positively charged exposed heme edge region on the front side of horse cytochrome c and Paracoccus cytochrome c2 . In contrast, at low ionic strength, the positively charged cobalt phenanthroline ion interacts with the negatively charged back side of cytochrome c2, and at high ionic strength at a positively charged site on the front side of the cytochrome . With horse cytochrome c, over the ionic strength range studied, cobalt phenanthroline reacts only at a positively charged site which is probably not at the heme edge . These inorganic oxidants do not react at the relatively uncharged exposed heme edge sites on Azotobacter cytochrome c5 and Pseudomonas cytochrome c-551, but rather at a negatively charged site which is away from the heme edge . The results demonstrate that at least two electron-transferring sites on a single cytochrome can be functional, depending on the redox reactant used and the ionic strength . Electrostatic interactions between charge distributions on the cytochrome surface and the other reactant, or interactions involving uncharged regions on the protein(s), are critical in determining the preferred sites of electron transfer and reaction rate constants . When unfavorable electrostatic effects occur at a site near the redox center, less optimal sites at a greater distance can become kinetically important.

Microbiol Rev, 1989 Jun, 53(2), 186 - 209
Dimorphism in Histoplasma capsulatum: a model for the study of cell differentiation in pathogenic fungi; Maresca B et al.; Several fungi can assume either a filamentous or a unicellular morphology in response to changes in environmental conditions . This process, known as dimorphism, is a characteristic of several pathogenic fungi, e.g., Histoplasma capsulatum, Blastomyces dermatitidis, and Paracoccidioides brasiliensis, and appears to be directly related to adaptation from a saprobic to a parasitic existence . H . capsulatum is the most extensively studied of the dimorphic fungi, with a parasitic phase consisting of yeast cells and a saprobic mycelial phase . In culture, the transition of H . capsulatum from one phase to the other can be triggered reversibly by shifting the temperature of incubation between 25 degrees C (mycelia) and 37 degrees C (yeast phase) . Mycelia are found in soil and never in infected tissue, in contrast to the yeast phase, which is the only form present in patients . The temperature-induced phase transition and the events in establishment of the disease state are very likely to be intimately related . Furthermore, the temperature-induced phase transition implies that each growth phase is an adaptation to two critically different environments . A fundamental question concerning dimorphism is the nature of the signal(s) that responds to temperature shifts . So far, both the responding cell component(s) and the mechanism(s) remain unclear . This review describes the work done in the last several years at the biochemical and molecular levels on the mechanisms involved in the mycelium to yeast phase transition and speculates on possible models of regulation of morphogenesis in dimorphic pathogenic fungi.

J Gen Microbiol, 1989 Jun, 135 ( Pt 6), 1607 - 17
Influence of oestradiol on protein expression and methionine utilization during morphogenesis of Paracoccidioides brasiliensis; Clemons KV et al.; The temporal sequence of cytosolic protein expression during phase transition of Paracoccidioides brasiliensis was examined . Electrophoretic analysis of cytosol proteins by one-dimensional SDS-PAGE revealed numerous differences between the mycelial and yeast forms as well as alterations induced by 17 beta-oestradiol . Using either protein staining or fluorography of {35S}methionine-labelled proteins 30 phase-specific bands were detected, 12 mycelial-associated bands (range 30 to 140 kDa) and 18 yeast-associated bands (range 22 to 127 kDa) . In cells undergoing mycelial to yeast transition after a shift from 25 degrees C to 37 degrees C, the protein patterns showed a temporal progression toward the yeast profile with the accumulation of yeast bands prior to observable morphogenesis . Five novel protein bands (range 23 to 50 kDa) were detected by silver staining during transition . Treatment of temperature-shifted mycelial cultures with 2.6 x 10(-7) M-oestradiol altered observed profiles; 4 of 12 mycelial-associated bands were maintained whereas the appearance of the 5 novel transition bands and 9 of 18 yeast-associated bands was blocked or delayed . Analysis of {35S}methionine-labelled proteins revealed that oestradiol induced label uptake by mycelial cells, blocked the synthesis of a 92 kDa yeast-specific band 72 h into transition, and diminished label incorporation 120 h into transition . In conjunction with these steroid-induced alterations of protein expression, little or no morphological transformation occurred . These results support our hypothesis that, analogous to mammalian steroid receptor action, the functional responses of P . brasiliensis to oestradiol are related to regulation of protein expression, presumably mediated via a specific binding protein-ligand complex.

Arq Neuropsiquiatr, 1989 Jun, 47(2), 224 - 9
{Medullary paracoccidioidomycosis: report of a case}; Marchiori E et al.; Paracoccidioidomycosis of the central nervous system presents either as meningeal or pseudotumoral lesions . Although occurring more frequently in the brain and meninges, they can occasionally involve the spinal cord . A case of paracoccidioidomycosis in the cervical spinal cord is reported in this paper . Difficulties in establishing the etiologic diagnosis, the importance of radiologic examination of the thorax, and the treatment of the patient are commented . In an extensive review of the literature on the subject, only three other cases have been found, which are also discussed.

Rev Med Panama, 1989 May, 14(2), 112 - 5
{Infection caused by Paracoccidioides brasiliensis in people living in Cocle and Veraguas, Republic of Panamá}; de Martin MC et al.; Intradermal application of yeast phase paracoccidioidin antigen obtained by sonication and lysis from the fungus to 111 residents of Veraguas province (68 men and 43 women) and 130 Cocle province residents (57 men and 73 women) elicited a positive reaction in 9.01% (1/111) and 26.15% (34/30) of the Veraguas and Cocle residents respectively . Positive reactions ranged between papules of 5 and 27 mm in diameter, the majority being between 11 and 20 mm . Infection was more frequent in men (1:1.47 for Veraguas and 1:1.28 for Cocle) and in those involved in farming . A prevalence of 21.95% P . brasiliensis infection is demonstrated for the Republic of Panama.

Rev Inst Med Trop Sao Paulo, 1989 May-Jun, 31(3), 151 - 7
Chlamydospore formation by Paracoccidioides brasiliensis mycelial form; Franco M et al.; To investigate the role of some adverse environmental conditions in chlamydospore formation by the mycelial form of P . brasiliensis, we cultured four P . brasiliensis isolates (18, Bt4, 1183, Pb9) at 25 degrees C within solid agar medium either rich or poor in nutrients . Isolates 18 and 1183 were also cultured under anaerobiosis in a nitrogen atmosphere . Isolate 18 produced great number of terminal and intercalary chlamydospore after 7-10 days of culture in a medium poor in nutrients (2% agar with 0.1% dextrose and polypepton) . The three other isolates also produced chlamydospores under the same conditions, but in lower numbers . Chlamydospore production by isolate 18 was abolished when the fungus was cultured in two agar media rich in nutrients (brain heart infusion and potato dextrose agar) . Anaerobic incubation of isolate 18 under an atmosphere of N2 showed small mycelial outgrowth with numerous chlamydospores . At the electron microscopical level, the chlamydospores showed one or various nuclei and numerous mitochondria, indicating great potential for further development . Accordingly, chlamydospores produced multiple budding after only 24 h incubation at 35 degrees C . The results demonstrate that under adverse environmental conditions P . brasiliensis mycelial form produces chlamydospores within a short period of time.

Mycopathologia, 1989 Apr, 106(1), 53 - 8
Digestion of killed Paracoccidioides brasiliensis by neutrophils; Goihman-Yahr M et al.; We previously described an in vitro assay showing that neutrophils (PMNs) from patients with paracoccidioidomycosis (PARA) have a specific digestive deficiency against suspensions of live Paracoccidioides brasiliensis . We now report that this defect is equally detectable against autoclaved, but not Amphotericin B-killed P . brasiliensis . The use of autoclaved suspensions facilitates the use of our in vitro assay . It might allow the development of an in vitro intradermal test for digestion of fungi . Differential digestive ability of phagocytes against live (or autoclaved) and Amphotericin-B killed fungi is of conceptual interest . It may be relevant in understanding therapeutic effect of Amphotericin B.

Infect Immun, 1989 Mar, 57(3), 896 - 901
Production and characterization of murine monoclonal antibodies to Histoplasma capsulatum yeast cell antigens; Kamel SM et al.; Four monoclonal antibodies (MAbs) were produced by immunizing mice with a disrupted yeast cell homogenate of Histoplasma capsulatum . MAbs 1 and 2 reacted only with the yeast cell antigens of H . capsulatum and Blastomyces dermatitidis, whereas MAbs 3 and 4 showed broader cross-reactivity . MAb 3 cross-reacted with B . dermatitidis, Paracoccidioides brasiliensis, Sporothrix schenckii, and Candida albicans, and MAb 4 cross-reacted with B . dermatitidis, C . albicans, Coccidioides immitis, Aspergillus fumigatus, and Mycobacterium tuberculosis . All four MAbs exhibited unique specificity when reacted with three different strains of H . capsulatum (G217B, A811, and P-IN) . MAb 1 belonged to the IgG2b subclass, MAb 3 belonged to the IgG1 subclass, and MAbs 2 and 4 belonged to the IgG3 subclass . MAbs 1, 2, and 3 formed bands in the Western immunoblot assay; the two dominant distinct bands had apparent molecular masses of 72 and 62 kilodaltons.

Rev Inst Med Trop Sao Paulo, 1989 Mar-Apr, 31(2), 119 - 25
{Paracoccidioidomycosis and infection by the human immunodeficiency virus}; Pedro Rde J et al.; We present two cases of paracoccidioidomycosis, one occurring in an AIDS patient and the other in an HIV infected man . This is the first report of such association . The first patient, which was already followed for HIV infection (group IV-A) presented with high fever and hepatosplenomegaly . Plain X-ray, ultrasound and CT-scan of the abdomen showed solid nodules in the spleen, some of them with calcification . Both the direct smear and the culture of a bone marrow aspiration revealed Paracoccidioides brasiliensis . The patient died of acute disseminated Paracoccidioidomycosis . The second patient, a man anti-HIV seropositive presented with a mass on the right lower abdomen and inguinal region . A biopsy of the mass showed the association of Hodgkin's disease of the mixed cellularity type and paracoccidioidomycosis . With the expanding AIDS epidemic we believe this report emphasizes the need to consider Paracoccidioidomycosis in HIV infected persons in countries where this mycosis is endemic . We also suggest the inclusion of Paracoccidioidomycosis as a potential opportunistic infection in these areas.

Mycopathologia, 1989 Feb, 105(2), 71 - 4
Paracoccidioidomycosis in a patient with acquired immunodeficiency syndrome; Goldani LZ et al.; The authors report a case of paracoccidioidomycosis and other opportunistic diseases in a patient with acquired immunodeficiency syndrome.

Rev Inst Med Trop Sao Paulo, 1989 Jan-Feb, 31(1), 18 - 22
Inapparent lung involvement in patients with the subacute juvenile type of paracoccidioidomycosis; Restrepo A et al.; Three patients with the diagnosis of subacute juvenile paracoccidioidomycosis who, at the time of their first visit, had no signs or symptoms of lung involvement were studied . Initially the diagnosis was confirmed by the observation of P . brasiliensis in biopsy material obtained from clinically involved lymphadenopathies . The lung X-rays done in all patients, did not reveal pathologic changes, although it was possible to observe and isolate the fungus from sputum samples obtained from the three patients . This fact reinforces the pulmonary genesis of the mycosis and proofs the existence of a pulmonary primary infection, even in patients with the juvenile manifestations, in whom the lung component is obscured by the predominant lymph node involvement.

J Med Vet Mycol, 1989, 27(4), 243 - 52
Suppression of IgE antibody production against an unrelated antigen in experimental murine paracoccidioidomycosis; Amarante-Mendes JG et al.; A suppression of the IgE antibody response to ovalbumin was obtained in susceptible mice infected with Paracoccidioides brasiliensis yeast cells a few days prior to immunization with the former antigen plus adjuvant . A direct relationship between the number of injected fungi and the suppressive effect was established . When infection with a pathogenic isolate of P . brasiliensis (Pb 18) was compared to a non-pathogenic isolate (IVIC Pb267), the IgE anti-ovalbumin response was reduced by both . A similar effect was observed if mice were injected with dead yeast cells prior to immunization . Two strains of mice with completely opposite susceptibilities to infection with Pb18 cells (B10.A--susceptible and A/SN--resistant) both showed suppressed IgE anti-ovalbumin antibody production when infected 3 days prior to immunization . Injection of both strains of mice with P . brasiliensis antigen on the same day as immunization also had the same suppressive effect . These results suggest that the suppression of IgE response to an unrelated antigen in experimental murine paracoccidioidomycosis could be due to antigenic competition or to a suppressive component present in P . brasiliensis cells.

Braz J Med Biol Res, 1989, 22(2), 205 - 12
The possible role of circulating immune complexes in the deficiency of cell-mediated immunity in paracoccidioidomycosis; Chequer-Bou-Habib D et al.; 1 . The nature and extent of immune abnormalities was studied in 28 untreated patients with a chronic moderate form of paracoccidioidomycosis (PCM) . 2 . The patients presented hyporeactivity to skin tests, diminished lymphocyte transformation by mitogens such as phytohemagglutinin-P and concanavalin A and by Paracoccidioides brasiliensis protein antigen . They also presented peripheral blood leukocytosis but normal absolute numbers of T-cell and T-cell subsets . 3 . The patients had increased serum levels of C3d, as well as high levels of circulating immune complexes (CIC) detected by C1q-binding and protein A-binding assays . 4 . There was a significant negative correlation between lymphocyte transformation by mitogens and CIC levels which suggested that CIC may be involved in the genesis of the depressed cell-mediated immunity in PCM patients.

J Med Vet Mycol, 1989, 27(3), 141 - 8
Virulence of a variant of Paracoccidioides brasiliensis that exists in the yeast form at room temperature; Villar LA et al.; Paracoccidioides brasiliensis (ATCC 60885) produces conidia that, when plated in enriched media and incubated at 21-25 degrees C, give rise to a yeast-form variant (YRT) (ATCC 46678) . The virulence of this variant for BALB/c mice has been compared with that of the yeast and conidia produced by the parent isolate . Adult female mice were inoculated intravenously with 10(7) viable cells of YRT or yeast, or 5 x 10(5) viable conidia . Mice were weighed weekly, and three mice were sacrificed at 1, 2, 3, 4 and 8 weeks post-inoculation and their organs weighed and prepared for determination of viable counts (colony forming units; c.f.u.) . Animals infected with yeast cells or with conidia exhibited little weight loss in comparison with that seen in the YRT-infected mice which began to lose weight 1 week post-challenge . Organ weights increased in the YRT group, while the changes in the remaining two groups were minor . The c.f.u . from these organs also increased in the YRT-infected mice, whereas the infection in the yeast-and conidia-infected mice was self-limiting . A proportion (16.6%) of the YRT-infected animals died during the observation period . Conversely, no deaths occurred in the remaining groups . These results indicate that the YRT variant possesses increased virulence.

J Med Vet Mycol, 1989, 27(2), 71 - 82
Evaluation of the pathogenicity and immunogenicity of seven Paracoccidioides brasiliensis isolates in susceptible inbred mice; Singer-Vermes LM et al.; In this investigation the pathogenicity and immunogenicity of seven isolates of Paracoccidioides brasiliensis were compared . The pathogenicity of each isolate was determined by 50% lethal dose estimations and histopathology analysed during a 24-week-period . Four basic patterns of virulence could be defined after intraperitoneal infection of susceptible, genetically homogenous, B10.A mice, namely, slightly virulent (isolates Pb 265 and IVIC Pb 267), intermediate (isolates Pb 192, IVIC Pb 9 and Pb SN), virulent (isolate Pb 2052) and highly virulent (isolate Pb 18) . The granulomas induced by the individual isolates were similar although the evolution of inflammation and the organs affected varied according to the isolate used . The immunogenicity of each P . brasiliensis isolate was evaluated by measuring IgG titres with an ELISA method . The intermediate and the slightly virulent isolates induced weak antibody production whereas isolates Pb 18 and Pb 2052 induced stronger specific humoral responses . Differences in the kinetics of antibody production elicited by the different fungal isolates were also observed.

Mycopathologia, 1989 Jan, 105(1), 53 - 8
Histological and ultrastructural study of the inflammation evoked by Paracoccidioides brasiliensis antigen in previously immunized mice; Defaveri J et al.; Bentonite particles uncoated and coated with soluble antigen of Paracoccidioides brasiliensis (Pb) were intravenously injected into mice with and without previous immunization with Pb antigen . The inflammatory reaction around the bentonite emboli in small lung vessels was quantitated and morphologically studied by light and electron (EM) microscopy, 2 to 8 days after challenge . In control nonimmunized animals, coated and uncoated bentonite particles caused mild and nonspecific inflammation made up by macrophages . By EM, they formed loosely aggregated clusters with cytoplasm containing few organelles and borders without interdigitation . In immunized mice injected with coated bentonite particles, the inflammatory area was significantly greater than that in nonimmunized animals in all periods of study with maximum difference at day 2 . The inflammatory process at days 2 and 4 was characterized as mature granulomata, composed of macrophages with great number of organelles in the cytoplasm, large euchromatic nuclei and prominent nucleoli . Altogether these findings indicated a lesion with high metabolic activity, compatible with a granulomatous hypersensitivity reaction . At days 6 and 8, there was a change from mature to epithelioid granulomata, well demonstrated by EM which showed macrophages with characteristically interdigitated cytoplasmic borders . The results strengthen the importance of cellular immunity in the genesis of epithelioid granuloma in paracoccidioidomycosis and reinforce the usefulness of the present model in studies of the inflammatory cellular sequency and events in this mycosis.

J Med Vet Mycol, 1989, 27(3), 133 - 40
The interface of mycology and endocrinology; Stevens DA; Message molecules such as hormones or pheromones have been described in non-pathogenic fungi and largely control mating . In one instance, a fungal hormone has a sequence homologous with a mammalian hormone, binds to a mammalian receptor and produces a functional response in a mammalian cell . Some mammalian hormones have also been shown to inhibit or stimulate pathogenic and non-pathogenic fungi, although until recently fungal binders (receptors) had not been demonstrated and the concentrations studied were not physiologic . We studied Paracoccidioides brasiliensis, for human sex hormone binding and functional effect, because of the predominance of paracoccidioidomycosis in males . We found a cytosolic 17-beta-estradiol (E2) binding protein in mycelia with Kd = 13 nM and a capacity of 78 fmol mg-1 protein . E2 prevents mycelium-to-yeast or conidium-to-yeast conversion in vitro at close to physiologic concentrations . In vivo this action could inhibit or delay conversion to the pathogenic form, allowing sufficient time for stimulation of an effective immune response . More recent studies have demonstrated E2 block of fungal proteins associated with mycelium-to-yeast conversion, and effects on methionine uptake . In other studies, a progesterone binder in dermatophytes has been described . Progesterone inhibits the growth of these fungi, a finding which may also relate to the relative resistance of females to dermatophyte infection . A corticosteroid-binding protein has been described in Candida species.(ABSTRACT TRUNCATED AT 250 WORDS)

J Med Vet Mycol, 1989, 27(6), 407 - 12
Identification of antigenic polypeptides of Paracoccidioides brasiliensis by immunoblotting; Camargo ZP et al.; Exoantigens from cultures of Paracoccidioides brasiliensis were separated by SDS-PAGE and subsequently blotted onto nitrocellulose paper . Immunologically reactive components were detected by reaction with sera from patients with paracoccidioidomycosis (PCM) . Peroxidase-conjugated anti-human IgG was used as a detector antibody to reveal immunocomplexes . Anti-P . brasiliensis IgG reacted with four major components of 70, 52, 43 and 20-21 kDa and with eight minor components . The 43 kDa glycoprotein (gp) was the predominant IgG reactive antigen, recognized by 100% of the patient's sera . The antibody titre to the 70 kDa glycoprotein, recognized by 96% of the sera tested, decreased significantly in patients undergoing antimycotic therapy . Cross-reactions with the immunoblotting technique could be minimized by serum dilution . Both gp 43 and gp 70 can be considered to be markers for human PCM under the test conditions used in our immunological assay.

J Med Vet Mycol, 1989, 27(3), 149 - 58
In situ localization of T lymphocyte subsets in human paracoccidioidomycosis; Moscardi-Bacchi M et al.; Immunohistochemical techniques using monoclonal antibodies to T lymphocyte subsets were used to characterize granulomas caused by Paracoccidioides brasiliensis . Punch biopsies of skin or mucosa from eight patients and of lymph nodes from two patients with disseminated paracoccidioidomycosis were studied . The T lymphocytes were distributed either in a localized pattern related to epithelioid granulomas or in a diffuse arrangement, predominantly around the vessels . In the granulomas, T cells formed a peripheral mantle surrounding central aggregates of macrophages . The majority of lymphocytes were T-helper cells with few suppressor cells . In contrast, patients presented with a decreased number of peripheral T-helper lymphocytes and a corresponding decrease in helper-suppressor cell ratios . There was no clear-cut relationship between tissue helper-suppressor cell ratios and the level of cellular immunodepression of the patients . The lowest P . brasiliensis antibody titers were detected in patients with the highest tissue helper-suppressor cell ratios . The distribution pattern of T lymphocytes in P . brasiliensis granulomas, with a predominance of helper phenotype, suggests that these cells are actively involved in the disease process.

Rev Inst Med Trop Sao Paulo, 1989 Jan-Feb, 31(1), 40 - 3
{Agar gel microdiffusion reaction in the serologic diagnosis of paracoccidioidomycosis}; da Silva MI et al.; We used the micro- and macroimmunodiffusion test for the qualitative and quantitative measurement of anti--P . brasiliensis antibodies in serum of patients with paracoccidioidomycosis . All 103 paracoccidioidomycosis sera (100%) were positive in the microtest versus 87% positivity index in the macrotest . All 83 control sera from patients with other diseases were negative in both tests . Titers of the positive sera tended to be higher in the microtest, which revealed sharper and easier to read precipiting bands . Microimmunodiffusion is simple to be performed, requires a minimum amount of reagents and allows the simultaneous testing of 102 sera . It may replace the macrotest specially in laboratories dealing with great serologic routine.

J Med Vet Mycol, 1989, 27(2), 93 - 104
Hypersensitivity pneumonitis to Paracoccidioides brasiliensis antigens in mice; Defaveri J et al.; Hypersensitivity pneumonitis was induced in mice immunized with Paracoccidioides brasiliensis and challenged, one week later, with soluble (SPbAg) or particulate (PPbAg) antigen (formalin-killed yeast cells), administered by the intratracheal route . Between 24 and 48 h post-challenge, animals developed an interstitial and intra-alveolar pneumonitis . Macrophages and lymphocytes arranged focally into loose or mature granulomata were observed by light and electron microscopy . The distribution and fate of antigens was studied by immunofluorescence . Three hours after challenge with SPbAg the lungs showed linear fluorescent deposits, whereas after challenge with PPbAg the pattern was globular, corresponding to the particulate antigen . After 24 and 48 h, the pattern was diffuse and finely granular in both groups, with a decreasing number of animals showing detectable fluorescence . Immunization induced a positive footpad swelling test (FPT) in all animals . After pulmonary challenge, there was a significant decrease in FPT indices, interpreted as desensitization due to trapping of specifically sensitized lymphocytes in the lungs . In conclusion, immunization induced a marked cellular immune response, the inflammatory pattern and the tempo of the induced pneumonitis being compatible with delayed hypersensitivity in the lungs . Immunized mice also cleared the injected antigens rapidly . These data suggest that hypersensitivity pneumonitis may be an expression of pulmonary resistance to infection with P . brasiliensis.

Laryngoscope, 1988 Dec, 98(12), 1330 - 3
Trigeminal trophic syndrome; Arasi R et al.; Trigeminal trophic syndrome is an unusual condition also known as trigeminal neurotrophic ulceration or trigeminal neuropathy with nasal ulceration . The diagnosis is suggested when ulceration of the face, especially of the ala nasi, occurs in a dermatome of the trigeminal nerve that has been rendered anesthetic by a surgical or other process involving the trigeminal nerve or its central sensory connections . A history of paresthesias and self-induced trauma to the area further support the diagnosis . Neurological deficits causing trigeminal trophic syndrome may result from surgical trigeminal ablation, vascular disorders and infarction of the brainstem, acoustic neuroma, postencephalitic parkinsonism, and syringobulbia . The following etiologies of nasal ulceration should be excluded: postsurgical herpetic reactivation and ulceration, syphilis, leishmaniasis, leprous trigeminal neuritis, yaws, blastomycosis, paracoccidioidomycosis, lethal midline granuloma, pyoderma gangrenosum, Wegener's granulomatosis, and basal cell carcinoma . In the case reported here, the diagnosis of TTS was made primarily as a result of previous experience with the syndrome, underscoring the importance of physician recognition of this unusual disorder.

Infect Dis Clin North Am, 1988 Dec, 2(4), 913 - 24
Systemic fungal infections: diagnosis and treatment . I . Paracoccidioidomycosis; Sugar AM; Paracoccidioidomycosis is the designation for the disease caused by the dimorphic fungus Paracoccidioides brasiliensis . It is a disease of great importance in Latin America and constitutes an important diagnosis for consideration in patients from endemic areas who have pulmonary or systemic disease compatible with fungal infection . In the laboratory, basic work has uncovered important interactions between the fungus and the human sex steroid system . These studies may have important implications in our approach to this and other infectious diseases.

What Is Bioreactor?, What Is Botulism?, What Is Genetics?, What Is Pcr?, What Is Biofilm?, n, Bacterium, o, Bacteria, r, Microbe, i, Microbes, n, Microbiology, a, Haemophilus, r, Candida tropicalis, i, Biofilms, a, Biodegradation, s, Clostridia, n, Bacillus subtilis, n, Pseudomonas, a, Salmonella, s, Yeasts, r, Escherichia coli, o, Pasteurella, r, Streptococcal, e, Bacteria, n, Cell suspensions, c, Microbial, o, Eubacter, i, Enterobacters, n, Microbial, r, Bacillus, a, Lactobacillus, i, Bacteriological

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005