Tipos de estacas e substratos na propagação vegetativa da menta (Mentha arvensis L. ) / Types of cuttings and substrates in the vegetative propagation of mint (Mentha arvensis L. )

O trabalho teve como objetivo avaliar a propagação vegetativa da menta utilizando diferentes tipos de estacas e substratos. O experimento foi conduzido no Horto de Plantas Medicinais da Unimontes, campus Janaúba - MG. O delineamento experimental utilizado foi o inteiramente casualizado, em esquema fatorial 2 x 4 (dois tipos de estacas e quatro diferentes substratos) com quatro repetições, sendo cada parcela representada por seis estacas. Foram analisadas as variáveis comprimento de parte aérea e de raízes, massa seca de parte aérea e de raízes e número total de brotações formadas por planta. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste de Scott-Knott a 5% de probabilidade. A interação entre os fatores estacas e substratos não foi significativa para as variáveis estudadas, passando-se a estudar o efeito isolado de cada fator. A propagação de Mentha arvensis L. pode ser realizada tanto por estacas apicais como medianas, utilizando o substrato solo + areia + esterco bovino (2:1:1) para a produção de mudas de qualidade.

The purpose of the study was to evaluate the vegetative propagation using different types of mint cuttings and substrates. The experiment was conducted in the Garden of Medicinal Plants of Unimontes, in Janaúba - MG. The experimental design was completely randomized (CRD) in 2 x 4 factorial schemes (two types of poles and four different substrates) with four replications and each plot was represented by six cuttings. The variables analyzed were the length of the shoots and roots, the dry matter of the shoots and roots and the total number of shoots per plant. The data were subject to ANOVA and the means were compared by Scott-Knott's test at 5% of probability. The interaction among stem cuttings and substrates was not significant for the variables studied, thus, the isolated effect of each factor was studied. The propagation of Mentha arvensis L. can be performed either by apical cuttings as medians, using the substrate soil + sand + manure bovine (2:1:1) for the production of quality seedlings.

Antifungal activity of tri- and tetra-thioureido amino derivatives against different Candida species.

The in vitro antifungal activity of six thioureido substituted amines (P1-P6) was evaluated against Candida species, including Candida albicans, C. glabrata, C. krusei and C. parapsilosis. These tri- and tetra-thioureido amino derivatives with different methylation levels were synthesised through easy synthetic routes to evaluate their antifungal properties against Candida species. Among all studied derivatives, the tri-(2-thioureido-ethyl)-amine (P1) was the most active compound inhibiting C. albicans and C. glabrata at a concentration of 0.49 µg ml(-1); P3, the N,N',N'',N'''-hexamethyl-derivative, also showed inhibitory activity against C. albicans and C. glabrata, but in higher concentrations (250 µg ml(-1) ). The N,N',N'',N'''-tetramethylated amine (P5) only inhibited the growth of C. glabrata, but its corresponding N,N',N'',N'''-octamethyl derivative (P6) was also active against C. glabrata (125 µg ml(-1)) and it was the only compound active against C. parapsilosis. P2 and P4 showed no significant antifungal activity. The structure-activity relationship of the thioureido-substituted derivatives indicates that the molecular branching and the alkylation levels can influence the antifungal activity. This study demonstrated that thioureido derivatives exhibited significant antifungal activity against Candida species and that they can be considered as a very promising bioactive lead compound to develop novel antifungal agents.

Isolation of clinically relevant fungal species from solid waste and environment of dental health services.

AIMS: This study was undertaken to detect, identify and determine antifungal susceptibility of yeast strains isolated from dental solid waste and to evaluate airborne fungi in the Brazilian dental health care environment and in the waste storage room. METHODS AND RESULTS: A group of 17 yeast strains were identified by macroscopic and microscopic characteristics, API 20C Aux system and Multiplex PCR. All 104 airborne fungal colonies were identified by macroscopic and microscopic morphology. The CLSI broth microdilution method was utilized as the susceptibility test. Candida parapsilosis was the prevailing yeast species recovered from waste, followed by Rhodotorula glutinis. Three strains of Candida guilliermondii presented minimal inhibitory concentration values considered to be susceptible dose dependent (2 µg ml(-1)) to voriconazole. Of all airborne fungal species, 69% were recovered from the waste storage room and 31% were recovered from the clinical/surgical environment. Most of them were identified as Cladosporium spp. CONCLUSIONS: These findings reinforce the potential risk of waste handling and point out the need for safe management to minimize the spread of these agents to the environment. Filamentous fungi isolation in almost all sampled environments indicates that a periodic monitoring of airborne microbiota in the dental health care service environment is required. SIGNIFICANCE AND IMPACT OF THE STUDY: The survival of yeast strains for 48 h suggests that dental waste should be carefully controlled and monitored.

Transcutaneous electrical nerve stimulation at both high and low frequencies activates ventrolateral periaqueductal grey to decrease mechanical hyperalgesia in arthritic rats.

Transcutaneous electric nerve stimulation (TENS) is widely used for the treatment of pain. TENS produces an opioid-mediated antinociception that utilizes the rostroventromedial medulla (RVM). Similarly, antinociception evoked from the periaqueductal grey (PAG) is opioid-mediated and includes a relay in the RVM. Therefore, we investigated whether the ventrolateral or dorsolateral PAG mediates antinociception produced by TENS in rats. Paw and knee joint mechanical withdrawal thresholds were assessed before and after knee joint inflammation (3% kaolin/carrageenan), and after TENS stimulation (active or sham). Cobalt chloride (CoCl(2); 5 mM) or vehicle was microinjected into the ventrolateral periaqueductal grey (vlPAG) or dorsolateral periaqueductal grey (dlPAG) prior to treatment with TENS. Either high (100 Hz) or low (4 Hz) frequency TENS was then applied to the inflamed knee for 20 min. Active TENS significantly increased withdrawal thresholds of the paw and knee joint in the group microinjected with vehicle when compared to thresholds prior to TENS (P<0.001) or to sham TENS (P<0.001). The increases in withdrawal thresholds normally observed after TENS were prevented by microinjection of CoCl(2) into the vlPAG, but not the dlPAG prior to TENS and were significantly lower than controls treated with TENS (P<0.001). In a separate group of animals, microinjection of CoCl(2) into the vlPAG temporarily reversed the decreased mechanical withdrawal threshold suggesting a role for the vlPAG in the facilitation of joint pain. No significant difference was observed for dlPAG. We hypothesize that the effects of TENS are mediated through the vlPAG that sends projections through the RVM to the spinal cord to produce an opioid-mediated analgesia.

In vitro studies of anticandidal activity of goniothalamin enantiomers.

AIMS: The antifungal activity of (R)-goniothalamin (1) and (S)-goniothalamin (ent-1) was evaluated against six Candida species. The in vitro effect of these compounds on yeast adhesion to human buccal epithelial cells (BEC) and Candida albicans and C. dubliniensis biofilms progression were also investigated. METHODS AND RESULTS: Yeast susceptibility was evaluated by broth microdilution assay and showed that ent-1 exhibited higher potency against all fungal clinical isolated when compared to compound 1. Compounds 1 and ent-1 were as potent as fluconazole in inhibiting the adhesion of C. albicans and C. dubliniensis to BEC. XTT-reducing assay and scanning electron microscopy revealed that 1 and ent-1 were twice as potent as fluconazole in the inhibition of yeast biofilms progression. CONCLUSIONS: Our findings indicate that compounds 1 and ent-1 are potent anticandidal agents. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights goniothalamin enantiomers as promising lead compounds for the design of new antifungal with inhibitory activity on yeast adhesion and biofilm progression.

Biofilm formation and effect of caspofungin on biofilm structure of Candida species bloodstream isolates.

Candida biofilms are microbial communities, embedded in a polymeric matrix, growing attached to a surface, and are highly recalcitrant to antimicrobial therapy. These biofilms exhibit enhanced resistance against most antifungal agents except echinocandins and lipid formulations of amphotericin B. In this study, biofilm formation by different Candida species, particularly Candida albicans, C. tropicalis, and C. parapsilosis, was evaluated, and the effect of caspofungin (CAS) was assessed using a clinically relevant in vitro model system. CAS displayed in vitro activity against C. albicans and C. tropicalis cells within biofilms. Biofilm formation was evaluated after 48 h of antifungal drug exposure, and the effects of CAS on preformed Candida species biofilms were visualized using scanning electron microscopy (SEM). Several species-specific differences in the cellular morphologies associated with biofilms were observed. Our results confirmed the presence of paradoxical growth (PG) in C. albicans and C. tropicalis biofilms in the presence of high CAS concentrations. These findings were also confirmed by SEM analysis and were associated with the metabolic activity obtained by biofilm susceptibility testing. Importantly, these results suggest that the presence of atypical, enlarged, conical cells could be associated with PG and with tolerant cells in Candida species biofilm populations. The clinical implications of these findings are still unknown.

Curcumin as a promising antifungal of clinical interest.

OBJECTIVES: The antifungal activity of curcumin was evaluated against 23 fungi strains and its in vitro inhibitory effect on the adhesion of Candida species to human buccal epithelial cells (BEC) was also investigated. METHODS: The antifungal susceptibility was evaluated by broth microdilution assay following the CLSI (formerly the NCCLS) guidelines. The inhibitory effect of curcumin on the cell adhesion was performed with Candida species and BEC. RESULTS: Paracoccidioides brasiliensis isolates were the most susceptible to curcumin while the growth of Aspergillus isolates was not affected. Curcumin was much more efficient than fluconazole in inhibiting the adhesion of Candida species to BEC, particularly those strains isolated from the buccal mucosa of AIDS patients. CONCLUSIONS: The lack of antifungal compounds with reduced side effects highlights the importance of studying natural products for this purpose. Curcumin was a more potent antifungal than fluconazole against P. brasiliensis, the causal agent of the neglected disease paracoccidioidomycosis. Curcumin dramatically inhibited the adhesion of Candida species isolated from AIDS patients to BEC, demonstrating that curcumin is a promising lead compound that warrants further investigation into its therapeutical use in immunocompromised patients.

Antifungal activities of compounds isolated from Piper abutiloides Kunth.

Piperaceae is a family of tropical plants known to have antifungal, antibacterial, tumour-inhibitory, antiviral, antioxidant, molluscicidal and leishmanicidal activities. In this work, extracts and fractions from aerial parts of Piper abutiloides (Piperaceae), a traditional medicinal plant, were evaluated against the fungal species Candida albicans, C. parapsilosis, C. krusei, C. glabrata, C. tropicalis, Cryptococcus neoformans and Sporothrix schenckii. The results have shown that the antifungal activity of this plant can be concentrated in the hexanic fraction after partitioning its hydroalcoholic extract between hexane and 90% aqueous methanol. The chromatographic fractionation of the bioactive part was monitored with a bioautographic assay using C. glabrata, and allowed the isolation of three antifungal compounds: pseudodillapiol, eupomatenoid-6 and conocarpan. These compounds presented different potencies against the fungi tested, with the strongest effect being observed for eupomatenoid-6 against C. glabrata, which presented a minimal inhibitory concentration value of 0.3 microg spot(-1). Conocarpan showed antifungal activity without apparent cytotoxic effect on normal human lymphocytes, as assessed by the proliferation assay with human peripheral blood mononuclear cells stimulated with phytohaemaglutinin. This work reveals for the first time the occurrence of these compounds in P. abutiloides and justifies further studies to clarify their mechanisms of action.

Antifungal suscepitibility profile of candida spp. oral isolates obtained from denture wearers / Perfil de sensibilidade antifungica de isolados de candida sp obtidos de usuários de prótese total

Denture stomatitis is an inflammatory condition that occurs in denture wearers and is frequently associated with Candida yeasts. Antifungal susceptibility profiles have been extensively evaluated for candidiasis patients or immunosupressed individuals, but not for healthy Candida carriers. In the present study, fluconazole, itraconazole, voriconazole, terbinafine and 5-flucytosin were tested against 109 oral Candida spp. isolates. All antifungal agents were effective against the samples tested except for terbinafine. This work might provide epidemiological information about Candida spp. drug susceptibility in oral healthy individuals.

A estomatite protética é uma condição inflamatória que ocorre em usuários de prótese total e está frequentemente associada a leveduras do gênero Candida, Os perfis de suscetibilidade a antifúngicos têm sido extensivamente estudados em pacientes com candidíase ou em indivíduos imunossuprimidos, mas não em portadores sadios de Candida. No presente estudo, fluconazol, itraconazol, voriconazol, terbinafina e 5-flucitosina foram testados contra 109 isolados orais de Candida spp. Todos os agentes antifúngicos mostraram-se eficazes contra as amostras avaliadas, exceto a Terbinafina. O presente trabalho pode fornecer dados epidemiológicos com relação à susceptibilidade a antifúngicos de Candida spp em indivíduos com saúde oral.

Antifungal suscepitibility profile of candida spp. oral isolates obtained from denture wearers.

Denture stomatitis is an inflammatory condition that occurs in denture wearers and is frequently associated with Candida yeasts. Antifungal susceptibility profiles have been extensively evaluated for candidiasis patients or immunosupressed individuals, but not for healthy Candida carriers. In the present study, fluconazole, itraconazole, voriconazole, terbinafine and 5-flucytosin were tested against 109 oral Candida spp. isolates. All antifungal agents were effective against the samples tested except for terbinafine. This work might provide epidemiological information about Candida spp. drug susceptibility in oral healthy individuals.

Antifungal activity of the amyrin derivatives and in vitro inhibition of Candida albicans adhesion to human epithelial cells.

AIMS: The antifungal activity of amyrin pentacyclic triterpene and 15 synthetic derivatives was evaluated against Candida species. Additionally, inhibition of adhesion of Candida albicans to human epithelial cells in vitro was determined. METHODS AND RESULTS: Esterification of alpha- and beta-amyrin with a variety of acyl chlorides produced a series of analogue derivatives. These substances were synthesized to evaluate the antifungal properties against Candida species. Among the 15 derivatives, alpha- and beta-amyrin formiate (2) and alpha- and beta-amyrin acetate (3) were the most active, inhibiting all the Candida species tested in concentrations that ranged from 30 to 250 microg ml(-1). alpha- and beta-amyrin formiate inhibited the adhesion ability of C. albicans to buccal epithelial cells (BEC) in 65.3%. CONCLUSIONS: alpha- and beta-amyrin formiate and alpha- and beta-amyrin acetate derivatives exhibited potential antifungal activity against Candida spp. and amyrin formiate showed inhibition of the adhesion ability of C. albicans to buccal epithelial cells. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated that two derivatives of amyrin pentacyclic triterpene exhibited significant antifungal activity against Candida species. Additionally, alpha- and beta-amyrin formiate was as effective as fluconazole in inhibiting the adhesion of C. albicans to buccal epithelial cells.

Redução do efeito analgésico da estimulação elétrica nervosa transcutânea de baixa freqüência em ratos tolerantes à morfina / Reduction in analgesic effect from low-frequency transcutaneous electrical nerve stimulation in morphine-tolerant rats

OBJETIVO: Investigar o efeito da TENS de baixa (10 Hz) e alta freqüência(130 Hz) aplicadas na pata inflamada do rato após tratamento crônico com morfina. MÉTODO: Foram utilizados 140 ratos Holtzman fêmeas, nos quais a carragenina (Cg 250 æg/0,1ml) foi administrada na pata posterior direita para a indução da inflamação. TENS de baixa e alta freqüência foi aplicada por 20 min, após 2 h e 30 min da Cg e seu efeito medido através do método de Randall-Selitto. O antagonista opióide Naltrexona (3mg/kg,sc), foi administrado 30 minutos antes da TENS para verificar a liberação de substâncias opióides endógenas. A tolerância foi obtida após administração da morfina (10 mg/kg,sc), duas vezes ao dia, durante sete dias. O tratamento com TENS de baixa e alta freqüência foi realizado no oitavo dia às 2 h e 30 min após Cg. A análise estatística foi feita pelo método da análise de variância ANOVA (One Way) seguido de um teste "post hoc" (Teste de Bonferroni), com nível de significância quando p < 0,05. RESULTADOS: TENS de baixa e alta freqüência inibiu em 100 por cento a hiperalgesia induzida pela Cg. Animais tratados previamente com naltrexona mostraram completa reversão da analgesia induzida pela baixa freqüência mas não pela alta freqüência. Após tolerância à morfina, os valores da TENS de baixa freqüência indicaram total ausência de analgesia, ao contrário da TENS de alta freqüência que induziu anti-hiperalgesia. CONCLUSÃO: Conclui-se que a atividade analgésica da TENS de baixa freqüência é reduzida após o desenvolvimento de tolerância a morfina.

OBJECTIVE: To investigate the effects of low (10 Hz) and high-frequency (130 Hz) transcutaneous electrical nerve stimulation (TENS) applied to inflamed paws of rats following chronic treatment with morphine. METHOD: 140 female Holtzman rats were utilized. Carrageenan (250 æg/0.1 ml) was administered to the right hind paws to induce inflammation. Two and a half hours after carrageenan injection, low and high frequency TENS was applied to the inflamed paw for 20 min, and its effect was measured via the Randall-Selitto method. The opioid antagonist naltrexone (3.0 mg/kg, subcutaneously) was administered 30 min before TENS, to verify the release of endogenous opioids. Morphine tolerance (10 mg/kg, subcutaneously) was induced by twice-daily injection over seven days. Low and high frequency TENS treatment was carried out on the eighth day, 2.5 hours after carrageenan injection. Statistical analysis was performed using one-way analysis of variance (ANOVA), followed by the post hoc Bonferroni test, with a significance level of p < 0.05. RESULTS: Both low and high frequency produced 100 percent inhibition of carrageenan-induced hyperalgesia. Naltrexone-treated animals showed complete reversion of analgesia induced by low but not high-frequency TENS. After attaining morphine tolerance, the low-frequency TENS values indicated complete absence of analgesia, whereas high-frequency TENS induced anti-hyperalgesia. CONCLUSION: The analgesic activity of low-frequency TENS is reduced following the development of morphine tolerance.

Estudo da confiabilidade da força aplicada durante a mobilização articular ântero-posterior do tornozelo / Reliability study on the force applied during anteroposterior mobilization of the ankle joint

INTRODUÇÃO: Os fatores que contribuem para a eficiência da manobra de mobilização articular são a intensidade e a reprodutibilidade da força aplicada durante a mobilização, por um ou por diferentes examinadores. OBJETIVO: Avaliar a confiabilidade intra e interexaminador da força de mobilização ântero-posterior da articulação do tornozelo, grau III e IV de Maitland. MÉTODO: Os dois tornozelos de 35 voluntários assintomáticos foram mobilizados por dois examinadores experientes com o procedimento. Uma miniplataforma de força foi posicionada sob a perna do voluntário a fim de captar as forças obtidas durante as manobras de mobilização e seu sinal foi convertido e armazenado em um software DasyLab4.0, o que permitiu o registro dos picos mínimo e máximo das curvas da força aplicada. A análise da confiabilidade foi realizada através do coeficiente de correlação intraclasse (CCI). Para avaliar o erro sistemático das medidas foi utilizado o teste t pareado e o teste t independente, para as condições intra e interexaminadores, respectivamente. O método de Bland e Altman avaliou os limites de concordância das medidas entre os examinadores. RESULTADOS: Os dados demonstraram alta confiabilidade intra-examinador. A confiabilidade inter-examinador foi baixa e moderada para a força máxima e mínima respectivamente, durante a mobilização ântero-posterior da articulação do tornozelo. CONCLUSÃO: Esses dados sugerem que a força aplicada durante a mobilização articular grau III e IV de Maitland no tornozelo, apresenta alta confiabilidade intra-examinador e baixa para interexaminador.

Virulence and molecular diversity in Colletotrichum graminicola from Brazil.

Genetic diversity among 37 isolates of the sorghum anthracnose pathogen Colletotrichum graminicola, from four geographically distinct regions of Brazil, was evaluated by RAPD and RFLP-PCR markers and virulence characters on a set of 10 differential sorghum genotypes. Twenty-two races were identified and race 13B was the most frequent, but present in only two regions. RAPD analysis revealed 143 polymorphic bands that grouped the isolates according to their geographic origin, but not by their virulence phenotypes. RFLP with HaeIII, MspI, HinfI, HhaI, HpaII, EcoRI, HindIII, PstI, RsaI, Taq alphaI, and AluI enzymes over ITS domains and 5.8 rDNA genes of C. graminicola did not show differences among the isolates, indicating high conservation of these restriction sites. Molecular polymorphism was observed among isolates belonging to the same race. No association between virulence phenotypes and molecular profiles was observed.

Chromoblastomycosis caused by Rhinocladiella aquaspersa.

An unusual case is presented featuring chromoblastomycosis lesions localised in three different sites. The patient was a 52-year-old male farm worker from Barra do Corda, State of Maranhão, Brazil, who had had the disease for 2 years. Physical examination revealed extensive plaques situated on the left leg, left arm, forehead, and on the left side of the face. Direct examination of biopsies showed numerous sclerotic cells. The fungus was recovered in culture and identified on the basis of the characteristic conidiation as Rhinocladiella aquaspersa.

Brain abscess caused by Cladophialophora (Xylohypha) bantiana in a renal transplant patient.

Infectious disease is the most significant cause of morbidity and mortality in allotransplantation because of heavy immunosuppression. Brain abscesses caused by melanized fungi have been found occasionally and are an example of this complication. In this paper, we describe a case in a 61-year-old black man, who received a cadaveric kidney transplantation in December 1993, followed by triple therapy with cyclosporine, azathioprine, and prednisone. The patient developed right hemiparesis at the beginning of April 1998. A computed tomography scan showed a mass in the left parieto-temporal region of the brain. The patient underwent surgery and a brown-colored encapsulated brain abscess was resected. Histology of the tissue revealed a large number of pigmented fungal hyphae. Culture in a Sabouraud dextrose medium with cyclohexamide and chloramphenicol at 25 degrees C resulted in the growth of dark-green colonies. The fungus identified was Cladophialophora bantiana, based on characteristic microscopic features and on growth at 40 degrees C. The abscess recurred in spite of treatment with fluconazole. The patient was submitted to a second brain surgical procedure and was treated with amphotericin B in addition to fluconazole. Ten days later the patient's blood cultures became positive for Escherichia coli. After 3 days the patient died due to septic shock.

Reduced production of hyperalgesic substances by mononuclear cells from aged rats incubated with carrageenan: role of interleukin 2 and prostaglandins.

OBJECTIVE AND DESIGN: To compare the production of hyperalgesic substances by cells from aged (A; 24-month) and juvenile (J; 2-month) rats. MATERIAL AND METHODS: 4 x 10(5) purified mononuclear cells from J and A were 2 h-stimulated (test) or not (control) by 250 microg lambda-carrageenan/well. Supernatants (0.1 ml) were intraplantarly (ipl) injected in rat paws and development of mechanical hyperalgesia, in grams, evaluated. Rat interleukin 2 (IL 2) and prostaglandin E2 (PGE2) were also assessed for hyperalgesia development. RESULTS: Test supernatants from A compared with J induced significantly less hyperalgesia (-56 +/- 8.1 and -88.4 +/- 4.6 g, respectively, p < 0.05, ANOVA t test). Local injection of a specific, but not a control, antiserum against IL 2 significantly blocked both pure IL 2- and stimulated supernatants-derived hyperalgesia. In contrast to PGE-like materials, IL 2 content in supernatants was compatible with hyperalgesia development. CONCLUSIONS: Hyperalgesia induced by test supernatants was significantly less intense when derived from aged animals. IL 2 may have accounted for such hyperalgesia.

Prevalence of Candida spp. in hospitalized patients and their risk factors.

A total of 133 Candida spp. strains originating from a group of 100 patients from Santa Casa de Misericórdia of Belo Horizonte, Brazil, between March 1995 and December 1996, were first identified and classified into six different species: Candida albicans (51%), C. tropicalis (33%), C. parapsilosis (8%), C. glabrata (5%), C. krusei (2%) and C. guilliermondii (1%). All C. albicans strains were serotyped and 55% of these were found to belong to serotype A and 45% belonging to serotype B. The medical records of each patient were examined to characterise and survey the main risk factors associated with them. Most of the patients were between 60- and 80-years-old, 53% were males and 47% were females. Most patients were from the intensive care unit (ICU). Only 10 patients were not exposed to antimicrobial agents and 72 patients were not prescribed antifungal agents. Forty patients showed no other clinical condition and all showed some underlying disease that justified hospitalization. Eighty-seven patients had undergone some invasive procedure and 31 patients had been submitted to two different procedures simultaneously.

Hyperalgesia and edema responses induced by rat peripheral blood mononuclear cells incubated with carrageenin.

The aim of this study was to verify the role played by mononuclear cells in an acute (nonimmune) inflammatory reaction. Mononuclear cells purified from rat peripheral blood were incubated for 1, 2, or 24 h with 100 or 250 microg/ml carrageenin (Cg). The resultant donor supernatant was injected into recipient rats to test its ability to induce hyperalgesia (reduction in threshold for paw pressure) and edema (increase in paw volume). Mononuclear cell supernatants (MnS) induced a significant time- and dose-dependent hyperalgesia and edema in rat paws, which reached a maximal effect at 3 h, lasted for 6 h, and returned to basal levels at 24 h of injection. Prostaglandins and cytokines (interleukin 1, 2, 6, 8, and tumor necrosis factor alpha) accounted for the hyperalgesia induced by MnS, as it was reduced (40 to 90%) by synthesis inhibitors such as indomethacin, dexamethasone, rolipram, and cyclosporin added to the cultures at a microgram dose-range. Edema was dependent on serotonin release in rat paws. These results indicate that mononuclear cells may be important contributors to acute inflammatory reactions, especially under those conditions where pain is an important component.

Use of RAPD (random amplified polymorphic DNA) to analyse genetic diversity of dematiaceous fungal pathogens.

Thirteen strains of chromoblastomycosis and phaeohyphomycosis etiologic agent fungi were obtained from different geographical origins. These strains were genotypically compared by means of the RAPD (Random Amplified Polymorphic DNA) technique. The data generated showed a high degree of polymorphism between distinct species and a low polymorphism between strains of the same species. The results generated by these tests were subjected to a numerical taxonomy analysis, using the unweighted pair-group method. A phenogram was constructed for the set of strains studied. Based on its structure, we concluded that genotypical data provide enough information to us to use the unweighted pair-group method to cluster the strains in accordance to their respective species. The phenogram grouped in a single branch the strains of Fonsecaea pedrosoi and F. compacta species, indicating a great similarity between these fungi, and suggesting that the classification as distinct species may not be appropriate for these species of the genus Fonsecaea.