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 co