Foxp3 expression in lesions of the different clinical forms of American tegumentary leishmaniasis.

As the diversity in clinical presentation of American tegumentary leishmaniasis (ATL) is determined mainly by the immune response of host, our aim was to evaluate the in situ expression of Foxp3 [marker of regulatory T (Treg) cell] in lesions of the different clinical forms of ATL. Foxp3(+) cells were observed in 39.5% (32/81) of the samples and the number of positive cells was low in all the clinical forms. Even presenting a significantly lower number of CD4(+) T cells, diffuse cutaneous leishmaniasis (DCL) showed a higher expression of Foxp3 when compared with localized cutaneous leishmaniasis (LCL) and mucocutaneous leishmaniasis (MCL). In LCL and MCL, the number of Foxp3(+) cells correlated positively with the number of apoptotic cells (active caspase-3(+) cells). A positive correlation was also observed between the expression of active caspase-3 and FasL in these clinical forms. Our data suggest that increased number of Treg cells may be associated to the hyporesponsiveness observed in DCL and also indicate that the apoptosis may be a possible mechanism of action of Foxp3(+) Treg cell in LCL and MCL. However, further studies are required to better understand the mechanism of action of Treg cell.

Human papillomavirus prevalence among women with cervical intraepithelial neoplasia III and invasive cervical cancer from Goiânia, Brazil.

This study estimated the prevalence and distribution of human papillomavirus (HPV) types among women with cervical intraepithelial neoplasia (CIN) grade III and invasive cervical cancer from Goi s (Brazil Central Region). Seventy-four cases were analyzed and consisted of 18 CIN III, 48 squamous cell carcinomas, 4 adenocarcinomas, 1 adenosquamous carcinoma and 3 undifferentiated carcinomas. HPV-DNA sequences were examined in formalin-fixed and paraffin-embedded tissues using primers from L1 region GP5+/GP6+. Polymerase chain reaction products were typed with dot blot hybridization using probes for HPV 16, 18, 31, 33, 45, 54, 6/11, 42/43/44, 51/52, 56/58. The prevalence of HPV was estimated to be 76% (56/74). HPV 16 was the most frequently found type, followed by HPV 33, 18 and 31. The prevalence of untyped HPV was 6%; 79% percent of the squamous cell carcinoma cases and 61% percent of the CIN III were positive for HPV and the prevalence rate of HPV types was the same for the total number of cases. According to other studies, HPV type 16 is the most prevalent virus in all Brazilian regions, but there is variation regarding to other types. Type 18 is the second most prevalent HPV in North, Southeast and South Brazil regions and types 31 and 33 are the second most prevalent HPV in Northeast and Central Brazil, respectively.

Human papillomavirus prevalence among women with cervical intraepithelial neoplasia III and invasive cervical cancer from Goiânia, Brazil

This study estimated the prevalence and distribution of human papillomavirus (HPV) types among women with cervical intraepithelial neoplasia (CIN) grade III and invasive cervical cancer from Goiás (Brazil Central Region). Seventy-four cases were analyzed and consisted of 18 CIN III, 48 squamous cell carcinomas, 4 adenocarcinomas, 1 adenosquamous carcinoma and 3 undifferentiated carcinomas. HPV-DNA sequences were examined in formalin-fixed and paraffin-embedded tissues using primers from L1 region GP5+/GP6+. Polymerase chain reaction products were typed with dot blot hybridization using probes for HPV 16, 18, 31, 33, 45, 54, 6/11, 42/43/44, 51/52, 56/58. The prevalence of HPV was estimated to be 76 percent (56/74). HPV 16 was the most frequently found type, followed by HPV 33, 18 and 31. The prevalence of untyped HPV was 6 percent; 79 percent percent of the squamous cell carcinoma cases and 61 percent percent of the CIN III were positive for HPV and the prevalence rate of HPV types was the same for the total number of cases. According to other studies, HPV type 16 is the most prevalent virus in all Brazilian regions, but there is variation regarding to other types. Type 18 is the second most prevalent HPV in North, Southeast and South Brazil regions and types 31 and 33 are the second most prevalent HPV in Northeast and Central Brazil, respectively

[Evaluation of secondary bacterial infection's influence on the course of cutaneous leishmaniasis in Corte de Pedra, Bahia]. / Avaliação da influência da infecção bacteriana secundária na evolução da leishmaniose cutânea em Corte de Pedra, Bahia.

In order to study the prevalence of secondary bacterial infection in ulcerated lesions and its relationship to the healing process, 84 leishmaniotic patients were evaluated. Diagnosis of the secondary infection was made by bacterial aerobic culture of peripheral tissue specimen of the ulcer. All patients received antimonial therapy during 20 days and washed their ulcers with common soap. Cases were composed mainly of adolescent and adult farmer patients with single lesions. The evaluated ulcers were encountered on legs and feet in 47.6%. Secondary bacterial infection was found in 45/83 (54.2%), and was more frequent in lesions located below the knee. Staphylococcus aureus predominated (89%). The ulcers' healing process, evaluated in 79 patients one month after finishing treatment, was not influenced by the secondary bacterial infection.

Avaliação da influência da infecção bacteriana secundária na evolução da leishmaniose cutânea em Corte de Pedra, Bahia / Evaluation of secondary bacterial infection's influence on the course of cutaneous leishmaniasis in Corte de Pedra, Bahia

In order to study the prevalence of secondary bacterial infection in ulcerated lesions and its relationship to the healing process, 84 leishmaniotic patients were evaluated. Diagnosis of the secondary infection was made by bacterial aerobic culture of peripheral tissue specimen of the ulcer. All patients received antimonial therapy during 20 days and washed their ulcers with common soap. Cases were composed mainly of adolescent and adult farmer patients with single lesions. The evaluated ulcers were encountered on legs and feet in 47.6%. Secondary bacterial infection was found in 45/83 (54.2%), and was more frequent in lesions located below the knee. Staphylococcus aureus predominated (89%). The ulcers' healing process, evaluated in 79 patients one month after finishing treatment, was not influenced by the secondary bacterial infection.

Foram avaliados 84 pacientes leishmanióticos com o objetivo de verificar a prevalência de infecção bacteriana secundária das úlceras cutâneas e de estudar sua relação com a cicatrização das lesões. A infecção secundária foi diagnosticada mediante cultura bacteriana aeróbica de amostra de tecido da lesão. Todos os pacientes receberam tratamento antimonial durante 20 dias e fizeram lavagem da úlcera com água e sabão comum. A casuística foi composta principalmente de adolescentes e de adultos dedicados à lavoura, apresentando lesão única. Em 47,6%, as úlceras estudadas estavam localizadas nas pernas e nos pés. Verificou-se infecção secundária em 45/83 (54,2%), sendo mais freqüente nas lesões localizadas abaixo dos joelhos. O Staphylococcus aureus predominou (88,9%). A reepitelização completa das úlceras, avaliada em 79 pacientes um mês após o fim do tratamento, não foi influenciada pela infecção secundária.

Efeitos farmacológicos da suplementaçao dietética com arginina em ratos com tumor sólido de walker 256 / Pharmacological effects of arginine supplementation in rats walker 256 solid tumor

Os efeitos da suplementaçao com arginina na dieta de portadores de cancer sao controversos. O objetivo deste trabalho é avaliar os efeitos da suplementacao dietética com arginina no crescimento tumoral, disseminaçao metastática, metabolismo de aminóacidos, alteraçoes hematológicas, tempo de sobrevida e peso corporal de ratos Wistar com tumor sólido de Walker 256. Foram administradas soluçoes por via gástrica, contendo arginina nas concentraçoes de 4 por cento, 6 por cento e dieta padrao (controle). A suplementacao com arginina inibiu a desseminaçao de células tumorais no modelo experimental, embora crescimento tumoral nao tenha sido afectado significantemente. O tempo de sobrevida dos animais com tumor sólido nao foi afetado de forma significativa. Foi observado disminuiçao significativa de peso corpóreo após a administraçao da arginina a 6 por cento (p<0.05). O metabolismo dos aminóacidos foi estimulado após ingestao de suplementos com arginina, evidenciado pelos aumentos significativos dos níveis sanguíneos de arginina, ornitina, citrulina, prolina e histidina quando comparados ao grupo controle (p<0.001). O grau de anemia no tumor sólido foi menos severo nos grupos que receberam suplementaçao com arginina, evidenciado através do hemograma (p<0.001) e da análise citológica. Os resultados sugerem que a suplementaçao dietética com arginina a 6 por cento apresentou algum efeito benéfico no tumor sólido de Walker 256 que ultrapassa o efeito nutricional

Therapeutic trial in experimental tegumentary leishmaniasis caused by Leishmania (Leishmania) amazonensis. A comparative study between mefloquine and aminosidine.

One hundred and eighty-two male inbred C57/BL/6 mice were infected with 3 x 10(6) Leishmania (Leishmania) amazonensis promastigotes of the MHOM/BR/PH8 strain by means of a subcutaneous injection in the right ear. The animals were separated in three groups: 1) oral mefloquine hydrochloride treatment (16 mg/kg/day/10 days), 2) intramuscular aminosidine (Paromomycin) treatment (20 mg/kg/20 days) and 3) control. Twenty-six mice of each treated group were sacrificed, one at the end of treatment (nine weeks after inoculation), and one six weeks later (fifteen weeks after inoculation). Control Group animals were sacrificed at weeks six, nine and fifteen after inoculation. There was no significant difference between Group 1 (mefloquine) and Group 3 (control) subjects. Group 2 animals (aminosidine) presented the smallest differences of all, both at the end of the treatment and six weeks later. The histopato-logical parameters have shown the following findings: a) there was no significant difference between the mefloquine treated group and the control group; the group treated with aminosidine showed fewer of vacuolated macrophages than the control group, at week 9 (end of treatment). b) both at the end of treatment and six weeks later, evaluation of tissue necrosis and tissue fibrosis revealed no differences between the treated groups. It was found that six weeks after the end of treatment, mice in the control group presented significantly more severe degrees of fibrosis than mice in the other groups. It can be concluded that mefloquine showed limited therapeutic effect in this experimental model, whereas aminosidine had a significant effect. Nevertheless, neither of them resulted in cure of the lesions.

Action of pentoxifylline on experimental cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis.

In the animal model of leishmaniasis caused by Leishmania (Leishmania) amazonensis there is a complex mechanism of the host-parasite interaction. The present study was performed to interfere with the inflammatory reaction to the parasites, through immune modulation. Female C5BL/6 isogenic mice were used, some of which were inoculated on the right ear and others on the right footpad with 3.10(6) stationary phase promastigotes of the MHOM/BR/PH8 strain of L. (L.) amazonensis, and were allocated in three groups: the first received pentoxifylline 8mg/kg every 12 h, since the first day; the second one received the same dose since the 40th day of infection and a control group that did not receive any treatment. All the ears excised were analyzed to determine the variation in weight between both ears and for histopathological analyses. A quantification of the parasites was done using the limiting dilution assay. A significant reduction of the number of parasites, was observed among the animals treated which had an accordingly significant reduction on the weight of the ears. Pentoxifylline reduced the macrophages propensity to vacuolation and induced a more effective destruction of the parasites by these cells. Moreover, the group that began the treatment later did not show the same effectiveness.

[Pharmacological effects of arginine supplementation in rats with Walker 256 solid tumor]. / Efeitos farmacológicos da suplementação dietética com arginina em ratos com tumor sólido de Walker 256.

The effects of diet arginine supplementation for those with cancer are controversial. We evaluate the effects of dietetic supplementation with arginine over body weight, growth of tumor, metastatic dissemination, surviving time, amino acid metabolism, haematological changes of the rats with Walker 256 solid tumor. Intragastrical solutions with arginine at 4% and 6%, a standard diet (control) were administered to the animals. The supplementation with arginine was associated with a lower weight gain during the study period (p < 0.05). Surviving time of the rats with solid tumor did not vary significantly between the groups. The rate of metastase was lower in animals with Walker 256 solid tumor supplemented with arginine. The amino acid metabolism was estimulate in the animals after arginine supplementation at 4% and 6%, demonstrated by significant increases in blood levels of arginine, ornitine, citruline, proline and histidine when compared to the control group. Anaemia was less severe in the rats with Walker 256 solid tumor that received arginine supplementation. The results suggest that arginine 6% supplementation may have pharmacologic effect in rats with Walker 256 solid beyond the nutritional one.

[A comparative study between sodium stibogluconate BP 88R and meglumine antimoniate in the treatment of cutaneous leishmaniasis. I. The efficacy and safety]. / Estudo comparativo entre estibogluconato de sódio BP 88R e antimoniato de meglumina no tratamento da leishmaniose cutânea: I. Eficácia e segurança.

Efficacy and safety of meglumine antimoniate and sodium stibogluconate BP 88R were compared in cutaneous leishmaniasis treatment in Corte de Pedra, Bahia, an endemic area of leishmaniasis due to Leishmania (Viannia) braziliensis. An open trial was developed with one hundred twenty seven patients who were diagnosed based on clinical criteria and Montenegro's skin test. Fifty eight patients were treated with meglumine antimoniate and 69 received sodium stibogluconate. Both groups received 20 mg/Sbv/kg/day for 20 days. Patients were followed every ten days during treatment and every month thereafter for three months. Sixty two percent patients cured with meglumine antimoniate and 55% cured with sodium stibogluconate (p = 0.42). Headache was more frequent during the first half of treatment in patients receiving sodium stibogluconate (p = 0.026). During the second half, patients treated with sodium stibogluconate showed a greater frequency of myalgia/arthralgia (p = 0.004) and abdominal pain/anorexia (p = 0.004). Three patients treated with sodium stibogluconate had severe side effects.

[Mefloquine in the treatment of cutaneous leishmaniasis in an endemic area of Leishmania (Viannia) braziliensis]. / Mefloquina no tratamento da leishmaniose cutânea em uma área endêmica de Leishmania (Viannia) braziliensis.

The aim of this study was to evaluate the efficacy of mefloquine in the treatment of skin leishmaniasis in patients infected with Leishmania (Viannia) braziliensis at an endemic region. Mefloquine is an oral drug effective against malaria with a prolonged half-life, less toxicity and easier administration than pentavalent antimonials. At Corte de Pedra in the Southern litoral of Bahia State, two randomized groups of ten patients with leishmaniasis were treated. The first group was treated with oral mefloquine, 250 mg per day in a single dose for six days and repeated three weeks later. The second group received meglumine antimoniate (Glucantime), 20 mg/kg daily administered intravenously for 20 days. Only one patient in the group treated with mefloquine showed evidence of clinical success. During treatment, one patient with four lesions developed a new lesion. The other three patients with clinical leismaniasis did not show evidence of clinical success after nine weeks of treatment. The group treated with Glucantime showed evident clinical improvement of the skin lesions.

Clinical observations of unresponsive mucosal leishmaniasis.

We report the long-term clinical follow-up of two patients with unresponsive mucosal leishmaniasis due to Leishmania (Viannia) braziliensis from the Três Braços area in Bahia State, Brazil. Both were agricultural male workers with extensive upper respiratory mucosal involvement that was not cured with conventional and experimental therapy.

[Open therapeutic study with aminosidine sulfate in mucosal leishmaniasis caused by Leishmania (Viannia) braziliensis]. / Estudo terapêutico aberto com sulfato de aminosidine na leishmaniose mucosa causada por Leishmania (Viannia) braziliensis.

From September to November 1994. 21 patients with active mucosal leishmaniasis were treated with aminosidine sulphate 16 mg/kg/day by intramuscular injection for 20 days. They were principally adult male agricultural workers. Thirteen patients had not received specific treatment and eight had failed to respond to Glucantime therapy. Diagnosis was based on clinical and epidemiological observations, a search for the parasite, leishmanin skin sensitivity and indirect fluorescent antibody serological tests. Sixty seven percent of patients had leishmania parasites isolated from inoculated hamsters or visualized in imprints or histopathological sections. The mean follow-up period was 12.6 months. All patients completed treatment. Side effects were pain at the injection site (86%); mild proteinuria (24%), elevated serum creatinine (.5%) and subclinical bearing loss in one of two patients who did audiometric tests. Clinical cure was achieved in 48% and the accumulated relapse rate was 29% (4/14).

[Comparative study of meglumine antimoniate, pentamidine isethionate and aminosidine sulfate in the treatment of primary skin lesions caused by Leishmania (Viannia) braziliensis]. / Estudo comparativo entre antimoniato de meglumina, isotianato de pentamidina e sulfato de aminosidine, no tratamento de lesões cutâneas primárias causadas por Leishmania (Viannia) braziliensis.

With the aim of comparing the therapeutic efficacy, tolerability and toxicity of meglumine antimoniate, aminosidine sulphate and pentamidine isethionate, a field study was conducted on randomized treatment of patients with primary cutaneous leishmaniasis due to Leishmania (Viannia) braziliensis (L(V)b), in Corte de Pedra, BA, from October 1992 up to January 1993. Forty six patients were treated and distributed into three groups, two with 15 and one with 16 subjects. All patients were submitted to clinical examination, histopathological and immunological investigations, as diagnostic criterium. All patients were treated by intramuscular route. Group 1 received pentamidine 4 mg/kg/every 2 days, for 8 applications; Group 2 received aminosidine 20 mg/kg/day, for 20 days, and Group 3 received meglumine 10 mg Sbv/kg/day, for 20 days. Failure of therapy was defined as ulceration of the skin lesion four months after treatment. Such failure occurred in five cases as follows: two cases in patients of group 1 one case in patients of group 2, and two cases in group 3, after the first year of follow up. In the evaluation after three years we reviewed fifteen patients, five in each group; except for one in Group 3, all of them were cured. Statistical significance of the results between the three schedules used was not verified.

[Spontaneous healing of leishmaniasis caused by Leishmania viannia braziliensis in cutaneous lesions]. / Cura espontânea da leishmaniose causada por Leishmania viannia braziliensis em lesões cutâneas.

In field clinics in the communities of Três Braços and Corte de Pedra, Bahia, we have attended 1.416 patients with tegumentary leishmaniasis in fourteen years, the predominant species in transmission is Leishmania Viannia brasi liensis (LVB). Because of the danger of metastasis with this infection treatment was routinely recommended with Glucan-time. However sixteen patients refused injection therapy and six women were pregnant when seen and not treated. All patients were followed up in our clinic. All these patients closed their skin ulcers although one subsequently relapsed. Patients were followed up for variable periods (four to twelve years), after the diagnosis. In nine patients (40.9%) of the cohort, the time to healing after initiation of the lesion was calculated as six months of evolution. At twelve months, nineteen patients (86.3%) had complete healing of their lesions. In three patients an active lesion was present for longer than one year. The determinants of this variable natural evolution of human LVB lesion remains completely unknown. It is difficult for us to understand and compare the effects of therapeutic agents in mucocutaneous leishmaniasis.

[Information-related changes in attitude to mucocutaneous leishmaniasis of a population from an endemic region of the south of Bahia]. / Mudança do componente cognitivo da atitude de uma população de região endêmica do Sul da Bahia diante da leishmaniose tegumentar.

The mutilant nature of the mucosal form of leishmaniasis can cause alterations in personal relations. To identify the level of understanding of leishmaniasis and the psychological reaction to it patients and houseowners in an endemic area of Bahia were interviewed. Several false beliefs were identified and there was a strong tendency to reject mutilated patients among the population for a false fear of contagion. Possibly future clarification of the real nature of leishmaniasis could change such a negative attitude among the resident population.

Histological analysis and the pathogenesis of mucocutaneous leishmaniasis.

A series of over 400 well-documented biopsies of mucocutaneous leishmaniasis was evaluated to elucidate the histological processes associated with the elimination of parasites, and their correlation with the course of the disease. Non-specific inflammation was the most frequent and least effective response; its onset might be delayed, and in this event particularly the incidence of metastasis from skin to mucosa was high. Lysis of parasite-laden macrophages appeared to be the basic mechanism of parasite reduction, even when it was not overt. When it was acute the onset was usually rapid, and though it resulted in much tissue destruction the prognosis was generally better and mucosal metastasis rare. Lysis and non-specific inflammation both led to the formation of a post-necrotic type of granuloma, but reversion of the process was almost as common as progression. Ultimately a tuberculoid granuloma evolved and proceeded to resolution. In about 5 per cent of cases, macrophage activation appeared to bring about early resolution; neither reversion nor mucosal metastasis was seen.

Mucosal leishmaniasis in Brazil.

The clinical diagnosis and laboratory identification of Leishmania braziliensis braziliensis, a parasitic disease affecting the upper aerodigestive tract, is difficult. A retrospective computer-assisted analysis of patient records was done after examination of 58 patients with mucosal leishmaniasis in an endemic area of L. braziliensis braziliensis in Bahia, Brazil during January 1987. Biopsies of clinically active and clinically inactive mucosal patients were examined for parasites using routine hematoxylin and eosin histopathology and a new technique for rapid detection of Leishmania amastigotes using a genus-specific indirect immunofluorescent assay. No amastigotes were found in specimens from seven patients with clinically inactive mucosal disease using immunofluorescent monoclonal assay techniques, whereas specimens from seven out of 14 patients with clinically active mucosal disease were positive. These results suggest that the immunofluorescent antibody technique is markedly superior in identifying the intracellular amastigote in tissue sections of mucosal biopsies when compared to histopathology techniques or with other standard tests done in rural areas of Brazil. Various clinical and laboratory test data of the entire group of patients were examined and the efficacy of treatment evaluated. The median interval of time noted between cutaneous and mucosal disease was 4.5 years. Relapse was noted in 31% of patients treated with a low dose of meglumine antimoniate (10 mg per kg of body weight). Patients treated with a high dose of meglumine antimoniate (20 mg per kg of body weight) had a relapse rate of 27.3%. A chi-square statistical analysis revealed no significant difference (chi 2 = 0.049) between the two groups. Patients were considered cured if mucosal granulations were clinically absent after 4.6 years.

Recovery of Leishmania (Viannia) braziliensis from inoculated hamsters.

Leishmania (Viannia) braziliensis has never been isolated from wild animals although it is apparently capable of inducing infections in man, dogs, and donkeys. An analysis of the standard hamster culture system for analyzing infectivity of Leishmania sp. was undertaken. Results indicate that for L. (V.) braziliensis, routine cultivation of aspirates taken from the inoculation sites of 1-mo-infected hamsters should be undertaken. Moreover, in at least 1 of the 3 strains examined, isolation of the parasite was only achieved after 84 days of cultivation.