In vitro effects of lapachol and ß-lapachone against Leishmania amazonensis.

Leishmaniasis is a neglected disease that affects millions of people worldwide, and special attention should be given to treatment because the available drugs have limitations, which can lead to low therapeutic adherence and parasitic resistance. This study evaluated the activity of the bioactive naphthoquinones, lapachol and ß-lapachone, against Leishmania amazonensis. The cell alterations were evaluated in vitro on promastigote and amastigote forms. The lethal dose (LD50) at 24, 48, and 72 h on the promastigote's forms using lapachol was 75.60, 72.82, and 58.85 µg/mL and for ß-lapachone was 0.65, 1.24, and 0.71 µg/mL, respectively. The naphthoquinones significantly inhibited the survival rate of L. amazonensis amastigotes at 83.11, 57.59, and 34.95% for lapachol (82.28, 41.14, and 20.57 µg/mL), and 78.49, 83.25, and 80.22% for ß-lapachone (3.26, 1.63, and 0.815 µg/mL). The compounds on the promastigote's forms led to the loss of mitochondrial membrane potential, induced changes in the integrity of the membrane, caused damage to cells suggestive of the apoptotic process, and showed inhibition of tumor necrosis factor (TNF)-α and interleukin (IL)-6 production. The results showed that these naphthoquinones are promising candidates for research on new drugs with anti-Leishmania activity derived from natural products.

In vitro effects of lapachol and β-lapachone against Leishmania amazonensis

Leishmaniasis is a neglected disease that affects millions of people worldwide, and special attention should be given to treatment because the available drugs have limitations, which can lead to low therapeutic adherence and parasitic resistance. This study evaluated the activity of the bioactive naphthoquinones, lapachol and β-lapachone, against Leishmania amazonensis. The cell alterations were evaluated in vitro on promastigote and amastigote forms. The lethal dose (LD50) at 24, 48, and 72 h on the promastigote's forms using lapachol was 75.60, 72.82, and 58.85 μg/mL and for β-lapachone was 0.65, 1.24, and 0.71 μg/mL, respectively. The naphthoquinones significantly inhibited the survival rate of L. amazonensis amastigotes at 83.11, 57.59, and 34.95% for lapachol (82.28, 41.14, and 20.57 µg/mL), and 78.49, 83.25, and 80.22% for β-lapachone (3.26, 1.63, and 0.815 µg/mL). The compounds on the promastigote's forms led to the loss of mitochondrial membrane potential, induced changes in the integrity of the membrane, caused damage to cells suggestive of the apoptotic process, and showed inhibition of tumor necrosis factor (TNF)-α and interleukin (IL)-6 production. The results showed that these naphthoquinones are promising candidates for research on new drugs with anti-Leishmania activity derived from natural products.

Calophyllum brasiliense Modulates the Immune Response and Promotes Leishmania amazonensis Intracellular Death.

Calophyllum brasiliense is a plant from the Brazilian rain forests and has been used in folk medicine for the treatment of various diseases, including leishmaniasis. This infectious disease depends on the Leishmania sp. and the host immune response. C. brasiliense antileishmanial activity is well known, but the effects on immune response remain to be investigated. This study showed the leishmanicidal and immunomodulatory effects of a 30 µg/mL of hydroalcoholic extract of C. brasiliense in murine macrophages before and after Leishmania (Leishmania) amazonensis infection. The semiquantitative cytokine RNA expression was determined by RT-PCR and the anti-Leishmania activity was measured by infection index (IF). Hydroalcoholic extract of C. brasiliense reduced more than 95% of IF when used before and after Leishmania infection, with 3 and 24 h of treatment (p < 0.05). C. brasiliense inhibited or reduced significantly (p < 0.05) the TNF-α, IL-1ß, IL-18, and IL-10 mRNA expression. The antileishmanial and anti-inflammatory effects showed the potential of C. brasiliense as an alternative therapy for leishmaniasis and it must be investigated.

Phototoxic effect of aluminium-chlorine and aluminium-hydroxide phthalocyanines on Leishmania (l.) amazonensis.

This study investigated the activity of photosensitive phthalocyanines on promastigotes and amastigotes of Leishmania (L.) amazonensis. Aluminum phthalocyanine chloride (AlPcCl), Aluminum phthalocyanine hydroxide (AlPcOH) and zinc phthalocyanine (PcZn) were tested in the presence (matte red LED, potency of 2.5-2.3 µW for 30 min) and absence of light against L. amazonensis promastigotes and the parasite viability was evaluated after 24, 48 and 72 h. The amastigote forms were treated with AlPcCl and AlPcOH, following the same lighting protocols described for the promastigote forms, being evaluated after 24 h. Cytotoxicity to human erythrocytes and peritoneal macrophages was also evaluated. The results showed that AlPcCl and AlPcOH in the presence of light have antileishmania activity, with leishmanistatic effects on promastigotes and amastigotes of L. amazonensis, without causing cytotoxicity to peritoneal macrophages and erythrocytes. The concentrations that inhibited 50% of the promastigote forms after 24 h of light exposure were 0.21 ±â€¯0.08 µM for AlPcCl and 0.23 ±â€¯0.06 µM for AlPcOH. In 48 h and 72 h after the treatment, the IC50 of AlPcCl was 0.13 ±â€¯0.02 and 0.12 ±â€¯0.03 µM and for AlPcOH was 0.14 ±â€¯0.01 µM and 0.11 ±â€¯0.01 µM, respectively. PcZn showed no activity on promastigotes of L. amazonensis. This study showed a substantial photodynamic activity of the phthalocyanines AlPcCl and AlPcOH against intracellular amastigotes forms of L. amazonensis after irradiation, presenting IC50 values of 0.62 ±â€¯0.06 µM and 0.92 ±â€¯0.12 µM, respectively. These results support the possibility of using photodynamic therapy for the treatment of cutaneous leishmaniasis.

Effect of a topical formulation containing Calophyllum brasiliense Camb. extract on cutaneous wound healing in rats.

This study evaluated the wound healing effects of topical application of an emulsion containing the HPLC-standardised extract from Calophyllum brasiliense Cambess (Clusiaceae) leaves in rats. The macroscopic analysis demonstrated that the wounds treated with the C. brasiliense emulsion healed earlier than the wounds treated with emulsion base and Dersani®. The percentage of wound healing in the group treated with the C. brasiliense emulsion was significantly higher than in the other groups at 7 and 14 days. On day 14, the animals treated with the C. brasiliense emulsion exhibited a 90.67% reduction of the wound areas. The histological evaluation revealed that on day 21, the group treated with the C. brasiliense emulsion exhibited a significant increase in fibroblasts compared with the other groups. Thus, the C. brasiliense emulsion had healing properties in the topical treatment of wounds and accelerated the healing process.

Anti-tuberculosis neolignans from Piper regnellii.

The present study determined the anti-Mycobacterium tuberculosis activities of supercritical CO2 extracts, neolignans eupomatenoid-5 (1), conocarpan (4) and eupomatenoid-3 (7) and their derivatives (2, 3, 5, 6, and 8) from Piper regnellii, as well as their cytotoxicities. The supercritical CO2 extract from leaves was purified by chromatographic methods, yielding compounds (1), (4) and (7), which were identified by (1)H NMR and comparison with literature data. Anti-M. tuberculosis activity (H37Rv and clinical isolates) was evaluated using a resazurin microtiter assay plate (REMA) to determine the MIC. The cytotoxicity assay was carried out in macrophages J774G.8 by sulforhodamine B colorimetric assay. The supercritical CO2 extracts from leaves and stems, and compound (4) showed activity against M. tuberculosis (MIC 15.6 µg/ml). Compound (1) showed the best activity (MIC 1.9 µg/ml), with good SI. Compounds (7) and (8) showed low activity against M. tuberculosis H37Rv. The derivative compounds did not show increased anti-M. tuberculosis activity. This is the first report, to our knowledge, to describe neolignans from P. regnellii with activity against M. tuberculosis, and compound (1) is a potential candidate for future antituberculosis drugs.

Efficacy of components from leaves of Calophyllum brasiliense against Leishmania (Leishmania) amazonensis.

Leishmanicide potential of Calophyllum brasiliense leaves on promastigote and amastigote of Leishmania (Leishmania) amazonensis is evaluated. The LD(50) of dichloromethane extract and hexane fraction for promastigotes was respectively 40 microg/ml and 20 microg/ml. In mouse peritoneal macrophages infected with Leishmania amastigotes the Infection Index decreased respectively 100% and 84.2% in 80 microg/ml and 40 microg/ml concentrations of dichloromethane extract. Hexane fraction decreased infection index respectively by 98.7% and 91.3% within the same concentrations. It was found that pretreatment with dichloromethane extract or with hexane fraction of experimentally infected BALB/c mice decrease the volume of the lesions by L. (L.) amazonensis. Moreover, animals treated topically also revealed healing lesions. Besides, the parasite load in the animals' popliteal lymph nodes was significantly reduced in treated animals, showing that plant components actually control infection. Results show that crude extract and hexane fraction of C. brasiliense reveal a significant in vitro and in vivo leishmanicide activity.

American cutaneous leishmaniasis in dogs from an endemic urban area in Cianorte municipality, Paraná State, Brazil

American cutaneous leishmaniasis (ACL) was investigated in dogs from an urban endemic area in Cianorte, Paraná state, Brazil. Of 169 studied dogs, none presented suspected ACL lesions. Eleven animals (6.6 percent) had anti-Leishmania braziliensis antibodies (titers > 40) detected by the immunofluorescent antibody test (IFAT) while four (2.4 percent) showed L. braziliensis-complex DNA by the polymerase chain reaction (PCR). Although no associations were found between IFAT or PCR results and age, sex, origin, free-roaming animals or length of residence at the address, the majority of IFAT- or PCR-positive dogs were from the urban area of the city and were allowed to roam freely beyond their neighborhood. The presence of anti-Leishmania braziliensis antibodies and L. braziliensis-complex DNA in dogs from this urban area near a native-forest park indicates the importance of following up on these dogs to confirm the ACL diagnosis.

Cell migration induced by Leishmania (Leishmania) amazonensis, Leishmania (Leishmania) major and Leishmania (Viannia) braziliensis into the peritoneal cavity of BALB/c mice

In American cutaneous leishmaniasis, the initial infection phase is characterized by recruitment of neutrophils and monocytes. The migration of these cells in response to the presence of Leishmania in the peritoneum of affected animals remains unclear. The objective of this study was to investigate cell migration to the peritoneum of BALB/c mice after infection with Leishmania (Leishmania) amazonensis, Leishmania (Viannia) braziliensis and Leishmania (Leishmania) major. Initially, Leishmania spp. was intraperitoneally inoculated in five groups of six animals each and the cell migration was analyzed 0, 3, 6, 12, 24 and 48 hours after infection. Different cell counts were performed with a staining kit and showed a higher percentage of polymorphonuclear than mononuclear cells in all three species studied. The total cell count revealed peak migration in L. (L.) amazonensis and L. (L.) major at six hours, and in L. (V.) braziliensis at 12 hours. These results suggest that factors released from different cell types probably act by attracting polymorphonuclear cells, with the peak migration most likely depending on the species of Leishmania inoculated into the host.

Ocorrência de leishmaniose tegumentar em cães de área endêmica no Estado do Paraná / Occurrence of cutaneous leishmaniasis in dogs of endemic area, Paraná State

Investigou-se a infecção por parasitas do complexo Leishmania braziliensis em 50 cães da cidade de Maringá, PR, onde ocorreram casos humanos de leishmaniose tegumentar americana (LTA), entre agosto e dezembro de 2006. Os casos humanos de LTA ocorreram nos anos de 2003 a 2004. Nenhum animal apresentou lesão, mas 12 (24,0 por cento) tinham sorologia e/ou PCR positivas, e a positividade tanto da IFI como da PCR foi de 14,0 por cento. Trinta e cinco animais eram de residências, e oito deles (22,8 por cento) tiveram testes laboratoriais para LTA positivos. Os outros 15 eram cães errantes, sendo que quatro deles (26,7 por cento) apresentaram PCR positiva. A presença de animais com infecção assintomática por Leishmania em área onde a LTA é endêmica pode ser um indicador do potencial de sua transmissão para o homem e uma referência para a implantação de medidas de controle e prevenção da doença.

The infection by Leishmania braziliensis complex was studied in 50 dogs from Maringá, PR, where American Cutaneous Leishmaniasis (ACL) in humans have been detected. Data were collected from August to December 2006 in an area in which ACL cases in humans were reported from 2003 to 2004. Indirect immunofluorescent test (IIF) and polymerase chain reaction (PCR) were applied. No lesions were found in the animals, although 12 (24.0 percent) had positive IIF and/or PCR. Positiveness was 14.0 percent for IIF and PCR. Lab tests showed that eight (22.8 percent) out of the 35 home animals were ACL positive. The other 15 animals were stray dogs, out of which four (26.7 percent) were PCR positive. The appearance of asymptomatic Leishmania-infected dogs in an ACL endemic area may be an indicator of ACL transmission potential for humans and a reference for the establishment of control measures and disease prevention.

Direct detection of Mycobacterium bovis in bovine lymph nodes by PCR.

Thirty-five lymph node samples were taken from animals with macroscopic lesions consistent with Mycobacterium bovis infection. The animals were identified by postmortem examination in an abattoir in the northwestern region of state of Paraná, Brazil. Twenty-two of the animals had previously been found to be tuberculin skin test positive. Tissue samples were decontaminated by Petroff's method and processed for acid-fast bacilli staining, culture in Stonebrink and Lowenstein-Jensen media and DNA extraction. Lymph node DNA samples were amplified by PCR in the absence and presence (inhibitor controls) of DNA extracted from M. bovis culture. Mycobacterium bovis was identified in 14 (42.4%) lymph node samples by both PCR and by culture. The frequency of PCR-positive results (54.5%) was similar to that of culture-positive results (51.5%, P > 0.05). The percentage of PCR-positive lymph nodes increased from 39.4% (13/33) to 54.5% (18/33) when samples that were initially PCR-negative were reanalysed using 2.5 microl DNA (two samples) and 1 : 2 diluted DNA (three samples). PCR sensitivity was affected by inhibitors and by the amount of DNA in the clinical samples. Our results indicate that direct detection of M. bovis in lymph nodes by PCR may be a fast and useful tool for bovine tuberculosis epidemic management in the region.

Aspectos epidemiológicos da Leishmaniose Tegumentar Americana na região Noroeste do Estado do Paraná / Epidemiological aspects of American Cutaneous Leismaniasis in the northwest region of the state of Paraná

No Estado do Paraná, a primeira notificação de leishmaniose tegumentar americana ocorreu em 1917 e a partir de 1980 observou-se um aumento do número de casos, mantendo-se endêmica e acometendo pessoas de todas as faixas etárias e em ambos os sexos. Este estudo teve como objetivo realizar um levantamento epidemiológico sobre a ocorrência de LTA em pacientes atendidos no Laboratório de Ensino e Pesquisa em Análises Clínicas da Universidade Estadual de Maringá (LEPAC/UEM). Foi realizado um estudo retrospectivo e descritivo em base de dados secundário de 1986 a 2005, com 1656 pacientes segundo as variáveis: sexo, idade, ocupação, procedência, local de moradia, forma clínica e diagnóstico. A maioria dos pacientes era do sexo masculino (72,6%) e adquiriu a infecção no Estado do Paraná (97,8%), residia em área urbana (64,3%) dos quais 51,3% adquiriu a infecção durante atividades de lazer. O diagnóstico da maioria dos pacientes foi estabelecido nos dois primeiros meses de evolução das lesões (54,0%) e apresentando a forma cutânea da doença (88,9%). Uma parcela (34,4%) significativa dos pacientes que residiam em área rural adquiriu a infecção no domicílio ou peridomicílio. O estudo mostra a predominância da forma cutânea da leishmaniose tegumentar americana e sugere a atividade de lazer e o ambiente do domicilio como fatores preditivos importantes para a infecção.

The first notification of American cutaneous Leishmaniasis (LTA) in the state of Paraná, Brazil, occurred in 1917 and an increasing number of cases has been reported since 1980. This parasitic skin disease, spread by the bite of infected sandflies, is still an endemic problem, with recurrence in both sexes and in all age groups. The objective of this study was to perform a survey on the occurrence of American cutaneous Leishmaniasis in patients attended at the Teaching and Research Clinical Analysis laboratory at the State University of Maringá (LEPAC/UEM). A retrospective and descriptive study was carried out, based on secondary data (1986-2005) on 1656 patients, relating to their sex, age and occupation and the origin, clinical forms and positive diagnosis of the disease. Most of the patients were male (72.6%), lived in the urban area (64.3%) and acquired the infection during outdoor leisure activities (51.3%) in the State of Paraná (97.8%). The cutaneous form of the infection predominated (88.9%) and the diagnosis was made in the first two months of development of the lesions (54.0%). A significant part (34.4%) of the patients who lived in the rural area acquired the infection inside or close to their homes. The study shows the predominance of the cutaneous form of American cutaneous Leishmaniasis and suggests that leisure activity and housing conditions could be useful predictive factors for the infection.

Serological survey for Leishmania sp. infection in wild animals from the municipality of Maringá, Paraná state, Brazil

Leishmania sp. infection was investigated in wild animals from the Ingá Park, in the municipality of Maringá, Paraná state, Brazil, where American cutaneous leishmaniasis (ACL) is an endemic disease. Sixty-five mammals, comprising Didelphis albiventris, Cerdocyon thous, Lycalopex vetulus, Cebus apella, Dasyprocta azarae, Dasypus novemcinctus, Procyon cancrivorus and Nasua nasua, were captured. Blood samples were collected for parasite cultivation. Antibodies were investigated by direct agglutination test (DAT) using Leishmania (Viannia) braziliensis as antigen. Flagellates were observed in blood cultures of 14 (35.9 percent) Didelphis albiventris. Anti-Leishmania antibodies were detected in 31 (51.6 percent) specimens of Cerdocyon thous, Lycalopex vetulus, Cebus apella, Dasyprocta azarae, Procyon cancrivorus and Nasua nasua. These results suggest that Cerdocyon thous and Lycalopex vetulus (crab-eating fox), Cebus apella (capuchin monkey), Dasyprocta azarae (agouti), Procyon cancrivorus (crab-eating raccoon) and Nasua nasua (coati) play an important role in the ACL transmission cycle in the northwestern region of Paraná, Brazil.

Leishmania (Viannia) braziliensis: New primers for identification using polymerase chain reaction.

The objective of this study was to develop specific primers for Leishmania (Viannia) braziliensis species identification using PCR. The designed primers (LBF1 and LBR1) were evaluated for sensitivity and specificity using various L. (V.) braziliensis serodemes and various Leishmania species and also using Trypanosoma cruzi. A specific fragment of 536bp was detected from 50ng of DNA in a crude extract derived from L. (V.) braziliensis. The DNA fragment was not detected when DNA from other Leishmania species or from T. cruzi was used as template in the PCR. Furthermore, when tested with DNA from cutaneous leishmaniasis the designed primers and reaction gave positive results. Taking into consideration that the primers LBF1 and LBR1 could specifically identify L. (V.) braziliensis, they could be considered for use in L. (V.) braziliensis diagnosis and epidemiological studies.

Hepatitis C: serological and molecular diagnosis and genotype in haemophilic patients at the Regional Hemocenter of Maringá, Maringá PR Brazil.

Prior to the process of viral inactivation in the 1980s most haemophilic patients were infected with hepatitis C virus, currently one of the disease's main causes. The prevalence of infection by hepatitis C virus in haemophilic patients enrolled in the Hemocenter of the Regional University Hospital maintained by the State University of Maringá is evaluated from social and epidemiologic data and from serum and molecular tests. Possible association between haemophilia's type and severity, type of treatment, data of start of treatment and hepatic enzyme rates of Alanine Aminotransferase (ALT) and Aspartate Aminotransferase (AST) was verified. Sixty-nine haemophilic patients, 82.6% with haemophilia A and 17.4% with haemophilia B, in the 1 - 67 years age bracket, average age 22 years, were investigated. Whereas anti-HCV was investigated with third generation immunoenzymatic test and confirmed by HCV-RNA, LiPA technique determined genotypes. HCV-prevalence infection reached 37.7% and 21.7% respectively when antibody anti-HCV and HCV-RNA detection and associated significantly (p<0.05) with type of treatment, date of start of treatment and ALT and AST hepatic enzyme rates are taken into account. HCV genotype 1 predominated (53.3%) followed by genotype 3 (40.0%) and genotype 2 (6.7%). HCV predominance is high in haemophilic patients although infection has not been reported in patients under the age of 17.

American cutaneous leishmaniasis in horses from endemic areas in the north-central mesoregion of Paraná state, Brazil.

American cutaneous leishmaniasis (ACL) is an endemic disease in the north-west of Paraná state, Brazil, where it occurs in humans and dogs. Few studies have been undertaken on the occurrence of the disease in other domestic animals and horses. In this study we investigated the infection of horses by Leishmania in ACL-endemic rural areas. Direct agglutination test (DAT) and polymerase chain reaction (PCR) were employed. Fifty-five horses from rural areas in the municipalities of Doutor Camargo, Ourizona, São Jorge do Ivaí, Ivatuba and Santa Fé (Agua do O) were analysed. DAT-detected antibody titres varied from 10 to 640, and 42 (76.3%) animals presented titres > or =20. PCR analyses led to the detection of Leishmania (Viannia) DNA in the blood samples of three (7.1%) of 42 animals. The presence of anti-L. (V.) braziliensis antibodies and Leishmania (Viannia) DNA in horses from endemic areas suggests their participation in the ACL transmission cycle.

Detection of Leishmania (Viannia) DNA in blood from patients with American cutaneous leishmaniasis.

The aim of the present study was to investigate the presence of Leishmania (Viannia) subgenus DNA in peripheral blood from patients with cutaneous lesions due to American cutaneous leishmaniasis. The buffy coats from 68 blood samples were analyzed by polymerase chain reaction using the MP1L/MP3H primers. The parasite DNA was detected in 2 (3.4%) out of 59 patients who had amastigotes present in samples taken from lesions. The presence of Leishmania (Viannia) DNA in the blood of these patients indicates hematogeneous parasite dissemination.

Comparação entre imunofluorescência indireta e aglutinação direta para o diagnóstico sorológico da leishmaniose tegumentar americana em cães errantes / Comparison between indirect immunofluorescence and direct agglutination for the serologic diagnosis of American cutaneous leishmaniasis in stray dogs

Estudaram-se 92 cães errantes com o objetivo de comparar a imunofluorescência indireta (IFI) e a aglutinação direta (AD) na investigação da leishmaniose tegumentar americana (LTA) canina, no período de outubro de 1999 a novembro de 2001. Os animais foram examinados quanto à presença de lesões e submetidos à intradermorreação de Montenegro (IDRM) e à pesquisa de anticorpos anti-Leishmania por IFI e AD. A IFI apresentou sensibilidade de 78,9 por cento e especificidade de 93,8 por cento (título >40), e a AD sensibilidade de 57,9 por cento e especificidade de 68,8 por cento (título >80). Dois cães apresentavam lesão, mas a pesquisa do parasito foi negativa. A IDRM, realizada em 89 cães, foi positiva em três (3,4 por cento). Trinta e quatro cães (37,0 por cento) foram positivos para a IFI, 61 (66,3 por cento) para a AD e 69 (75,0 por cento) para a IFI e/ou a AD. Dos cães com a IFI positiva, sete eram de Floresta, 13 de Maringá, 12 de Leópolis, um de São Jorge do Ivaí e um (50,0 por cento) de Campo Mourão. Os resultados sugerem o contato prévio desses animais com o parasito.

Ninety-two stray dogs were studied in order to compare indirect immunofluorescence (IIF) and direct agglutination (DA) in the investigation of American cutaneous leishmaniasis (ACL), from October 1999 to November 2001. The dogs were evaluated for the presence of lesions, and submitted to the Montenegro skin test (MST) and the search for anti-Leishmania antibodies by IIF and DA. IIF showed 78.9 percent sensitivity and 93.8 percent specificity (titers >40), and DA presented 57.9 percent sensitivity and 68.8 percent specificity (titers >80). In two dogs there were lesions, but the parasite was not detected. The MST was positive in 3 (3.4 percent) out of 89 dogs. Thirty-four (37.0 percent) dogs showed anti-Leishmania antibodies through IIF, 61 (66.3 percent) through DA, and 69 (75.0 percent) through IIF and/or DA. Of the dogs with positive IIF, 7 came from the municipal areas of Floresta, 13 of Maringá, 12 of Leópolis, 1 of São Jorge do Ivaí and 1 (50.0 percent) of Campo Mourão. These data suggest previous contact of these animals with the parasite.

Immunomodulatory effect of Canova medication on experimental Leishmania amazonensis infection.

This study investigates the action of Canova medication (CM) on experimental infection by Leishmania (Leishmania) amazonensis, utilizing in vitro and in vivo assays. For the in vitro tests, Balb/c mouse peritoneal macrophages (5x10(5) cells in 500 microl of culture medium, supplemented with 10% fetal calf serum, penicillin (100 U/ml) and streptomycin (0.1 mg/ml) (were distributed in 24-well plates and CM was added at concentrations of 20 or 40%. Twenty-four hours later, the macrophages were infected with Leishmania amastigotes in culture medium. The effect of CM on macrophages leishmanicidal activity in 24 and 48 h cultures was evaluated by determining infection index and measuring nitric oxide (NO) production. The in vivo tests were performed in mice infected with 10(7)L. (L.) amazonensis promastigotes injected in to the right hind footpad (25 microl in phosphate buffered saline). The progression of the lesions was examined over a 9-week period by measuring footpad swelling, and the parasite load in regional lymph nodes and spleen. The in vitro results showed that at 40% CM reduced the infection index, and induced NO production in the elicited macrophages, which suggests that the inhibitory effect on infection index may be mediated by NO. In the in vivo infection, when administered, orally or subcutaneously in mice, CM reduced infection by L. (L.) amazonensis in the paws, resulting in smaller lesions. CM treatment also decreased parasite load in the regional popliteal lymph nodes and in the spleen. These results suggest that CM modulates experimental infection by L. (L.) amazonensis, controlling infection progression and limiting dissemination.

A leishmaniose tegumentar sob a perspectiva do paciente: resultado de uma prática educativa / Cutaneous leishmaniasis under the patient's perspective: an educational practice result

Foi realizado um estudo descritivo empregando a metodologia qualitativa entre portadores de leishmaniose tegumentar visando identificar a representatação que a doença tem para o seu portador, antes e depois do tratamento e intervenção educacional. O estudo foi realizado de setembro de 2000 a março de 2001 no Estado do Paraná, Brasil. Para a coleta dos dados foi realizada a entrevista semi-estruturada, sendo que os membros foram analisados pela técnica do discurso do sujeito coletivo. Foram destacadas as idéias centrais relacionadas com a etiologia da leishmaniose, sua transmissão/prevenção, gravidade, estigma, dificuldades do tratamento, sentimentos frente à doença e validade da educação em saúde.este estudo evidenciou que o acompanhamento dos pacientes e as medidas educacionais são de fundamental importãncia para que o grupo social assuma um maior controle sobre sua saúde, auxiliando sobremaneira no enfrentamento da doença e seu tratamento.

A descriptive study was carried out among cutaneous leishmaniasis patients using the qualitative methodology, aiming to identify the role that the disease has to its suffers both before and after treatment and educational intervention. The study was carried out from September 2000 to March 2001 in the state of Paraná, Brazil...