Ultrastructural study on the morphological changes to male worms of Schistosoma mansoni after in vitro exposure to allicin

INTRODUCTION: Garlic has a wide range of actions, including antibacterial, antiviral, antifungal, antiprotozoal and anthelmintic actions. This antiparasitic activity has been attributed to allicin, which is the main constituent of garlic. The present study aimed to investigate the in vitro activity of allicin on the tegument of adult Schistosoma mansoni worms using scanning electron microscopy. METHODS: Swiss Webster mice were infected with S. mansoni cercariae (100 per mouse) and sacrificed 50 days later to acquire the adult worms. These worms were collected by perfusion and placed in RPMI medium 1,640 at 37°C before transferring to RPMI media containing 0 (control), 5, 10, 15 and 20mg/mL of allicin, where they were incubated for 2h. The worms were fixed in 2.5% glutaraldehyde solution, washed twice, post-fixed in osmium tetroxide, washed twice and then dehydrated with ascending grades of ethanol. The samples were air-dried, mounted on stubs, gold coated in an ion sputtering unit and viewed using a scanning electron microscope. RESULTS: A concentration of 5mg/mL caused wrinkling in the tegument; a concentration of 10mg/mL resulted in changes to tubercles and loss or modification of spines. With 15 and 20mg/mL increasing damage to the tegument could be seen, such as vesicle formation and the presence of ulcers. CONCLUSIONS: These findings demonstrate the effect of allicin on adult S. mansoni worms and indicate that most of the changes occur at concentrations greater than that normally indicated for treatment.

Leishmanicidal activity and cytotoxicity of compounds from two Annonacea species cultivated in Northeastern Brazil

INTRODUCTION: Visceral leishmaniasis is endemic in 88 countries, with a total of 12 million people infected and 350 million at risk. In the search for new leishmanicidal agents, alkaloids and acetogenins isolated from leaves of Annona squamosa and seeds of Annona muricata were tested against promastigote and amastigote forms of Leishmania chagasi. METHODS: Methanol-water (80:20) extracts of A. squamosa leaves and A. muricata seeds were extracted with 10% phosphoric acid and organic solvents to obtain the alkaloid and acetogenin-rich extracts. These extracts were chromatographed on a silica gel column and eluted with a mixture of several solvents in crescent order of polarity. The compounds were identified by spectroscopic analysis. The isolated compounds were tested against Leishmania chagasi, which is responsible for American visceral leishmaniasis, using the MTT test assay. The cytotoxicity assay was evaluated for all isolated compounds, and for this assay, RAW 264.7 cells were used. RESULTS: O-methylarmepavine, a benzylisoquinolinic alkaloid, and a C37 trihydroxy adjacent bistetrahydrofuran acetogenin were isolated from A. squamosa, while two acetogenins, annonacinone and corossolone, were isolated from A. muricata. Against promastigotes, the alkaloid showed an IC50 of 23.3 µg/mL, and the acetogenins showed an IC50 ranging from 25.9 to 37.6 µg/mL; in the amastigote assay, the IC50 values ranged from 13.5 to 28.7 µg/mL. The cytotoxicity assay showed results ranging from 43.5 to 79.9 µg/mL. CONCLUSIONS: These results characterize A. squamosa and A. muricata as potential sources of leishmanicidal agents. Plants from Annonaceae are rich sources of natural compounds and an important tool in the search for new leishmanicidal therapies.

Enteric parasites in HIV-1/AIDS-infected patients from a Northwestern São Paulo reference unit in the highly active antiretroviral therapy era

INTRODUCTION: We describe the epidemiology of intestinal parasites in patients from an AIDS reference service in Northeastern São Paulo, Brazil. METHODS: Retrospective evaluation was done for all HIV-1/AIDS-positive patients whose Hospital de Base/São José do Rio Preto laboratorial analysis was positive for enteroparasites after diagnosis of HIV-1 infection, from January 1998 to December 2008. Statistical analysis was performed using the R statistical software version 2.4.1. The level of significance adopted was 5%. RESULTS: The most frequent protozoan was Isospora belli (4.2%), followed by Giardia lamblia (3.5%), Entamoeba coli (2.8%), and Cryptosporidium parvum (0.3%). Ancylostoma duodenale (1.4%) was the most frequently detected helminth, while Taenia saginata and Strongiloides stercoralis were found in 0.7% of the samples. The results showed that diarrhea was significantly associated with giardiasis and isosporiasis. However, no association was observed between CD4+ cell counts, viral load, and the characteristics of any particular parasite. CONCLUSIONS: Our data may be useful for further comparisons with other Brazilian regions and other developing countries. The data may also provide important clues toward improving the understanding, prevention, and control of enteric parasites around the world.

Clinical and laboratory alterations in dogs naturally infected by Leishmania chagasi

INTRODUCTION: Canine visceral leishmaniasis (CVL) is a zoonotic disease with different clinical manifestations. Parasitism often occurs in bone marrow, but changes have been observed in peripheral blood and serum biochemical parameters. The aim of this study was to evaluate the hematological and biochemical parameters in dogs naturally infected by Leishmania chagasi. METHODS: Eighty-five adult dogs of both sexes and various weights and ages from the Zoonosis Control Center of Fortaleza (CCZ) were used, selected by immunofluorescence assay (IFA) and considered positive with IFA titers greater than 1:40 and by visualizing amastigotes of Leishmania chagasi in smears obtained by bone marrow aspiration. The dogs (n = 85) were grouped according to clinical signs: negative (CN = 7), subclinical (CS = 10), and clinical (CC = 68). Blood samples were collected for determination of hematological and biochemical serum values. The experimental protocol was approved by the CEUA/UECE. RESULTS: The most frequent clinical signs were cachexia (77.9%), keratitis (61.8%), and lymphadenopathy (55.9%), and 86.8% of the animals showed more than one clinical sign characteristic of CVL. In CC were observed reductions in red blood cells (63%), hematocrit (72%), and hemoglobin (62%), as well as leukocytosis (33%), neutropenia (28%), thrombocytopenia (50%), uremia (45%), hyperproteinemia (53%, p<0.05), hypergammaglobulinemia (62%, p<0.01), and hypoalbuminemia (58%). CONCLUSIONS: Animals with the clinical form of the disease demonstrate hematological and biochemical changes consistent with anemia, uremia, hyperproteinemia, and hyperglobulinemia, which present themselves as strong clinical markers of visceral leishmaniasis associated with the signs previously reported.

Prevalence and risk factors for Toxoplasma gondii infection among pregnant and postpartum women attended at public healthcare facilities in the City of Niterói, State of Rio de Janeiro, Brazil

IntroductionTo determine the prevalence of immunoglobulin G (IgG) and immunoglobulin M (IgM) anti-Toxoplasma gondii antibodies among pregnant and postpartum women attended within the public healthcare system in Niterói, State of Rio de Janeiro, and to detect possible exposure factors associated with T. gondii infection in this population.MethodsIgM and IgG anti-T. gondii antibodies were investigated in 276 pregnant and 124 postpartum women by using the indirect immunofluorescence (IFAT) and immunoenzymatic assay (ELISA) techniques. The participants were selected by convenience sampling. All these 400 patients filled out a free and informed consent statement, answered an epidemiological questionnaire and were informed about the disease.ResultsAmong the 400 samples analyzed, 234 (58.5%) were reactive to IgG anti-T. gondii antibodies, according to the IFAT and/or ELISA assay. One pregnant woman was found to be reactive to IgM anti-T. gondii antibodies, with an intermediate IgG avidity test. Risk factor analysis showed that seropositivity was significantly associated (p<0.05) with age, contact with cats and presence of rodents at home. Through a logistic regression model, these associations were confirmed for age and contact with cats, while education at least of the high school level was found to be a protective factor.ConclusionsThe prevalence rate of IgG anti-T. gondii antibodies in the City of Niterói was high and the risk factors for infection detected after multivariate analysis were: age over 30 years, contact with cats and education levels lower than university graduate level.

Puerperal brain cryptococcoma in an HIV-negative woman successfully treated with fluconazole: a case report

Cryptococcus spp. cerebral abscesses are uncommon in immunocompetent subjects. The recommended induction treatment is the administration of amphotericin B plus flucytosine combined with resection for lesions ≥3cm. In this paper, we describe an HIV-negative woman diagnosed with a large cryptococcoma in the immediate postpartum period. The lesion was not resected, and due to amphotericin B intolerance, she received an extended course of fluconazole monotherapy. There was no disease recurrence during the 4 years of follow-up. The abrupt onset of her symptoms following delivery suggests that she developed a postpartum immune reconstitution syndrome. This case also demonstrates that in specific situations fluconazole monotherapy can be attempted in immunocompetent patients with cryptococcoma.

The epidemiology of envenomation via snakebite in the State of Piauí, Northeastern Brazil

INTRODUCTION: This communication describes a retrospective study of the epidemiology of snakebite cases that were recorded from 2007 to 2012 in the State of Piauí, northeastern Brazil. METHODS: Data were collected from the Injury Notification Information System database of the State of Piauí's Health Department. RESULTS: A total of 1,528 cases were identified. The cases occurred most frequently in rural areas between January and July. Victims were predominantly male farmers, and were typically 30-39 years old. Most victims were bitten on the foot, and received medical assistance within 1-3h after being bitten. CONCLUSIONS: The epidemiological profile of snakebites in the State of Piauí is similar to that in all of Brazil.

Opportunistic infections among individuals with HIV-1/AIDS in the highly active antiretroviral therapy era at a Quaternary Level Care Teaching Hospital

INTRODUCTION : In this study, clinical-laboratory and epidemiological characteristics are described for a group of 700 individuals with HIV (human immunodeficiency virus)/AIDS (acquired immunodeficiency syndrome) in the ART (antiretroviral therapy) era at a teaching hospital that provides a quaternary level of care, with an emphasis on opportunistic infections (OIs), co-infections and immune profile. METHODS : A retrospective cross-sectional study of AIDS cases was conducted from 1998 to 2008 by reviewing medical records from the Base Hospital/FUNFARME (Fundação Faculdade Regional de Medicina), São José do Rio Preto, São Paulo, Brazil. RESULTS: The individuals were 14 to 75 years of age, and 458 were males. Heterosexuals accounted for 31.1% of all patients. Eighty-three percent were on ART, and 33.8% of those presented difficulties with treatment adherence. OIs were analyzed from medical records, and Pneumocystis jiroveci pneumonia was the most prevalent, regardless of the LTCD4+ (TCD4+ Lymphocytes) levels. Individuals whose viral loads were ≥10,000 showed a 90% greater chance of neurotoxoplasmosis. For P. jiroveci pneumonia, neurotoxoplasmosis, esophageal candidiasis, pulmonary tuberculosis and neurocryptococcosis, the chances of infection were higher among patients with LTCD4+ levels below 200 cells/mm3. HIV/hepatitis C virus (HCV) and HIV/hepatitis B virus (HBV) co-infections were significantly associated with death. CONCLUSIONS : OIs remain frequent in the ART era even in populations where the access to medical care is considered satisfactory.

Molecular detection of Trypanosoma sp. and Blastocrithidia sp. (Trypanosomatidae) in phlebotomine sand flies (Psychodidae) in the Federal District of Brazil

Abstract: INTRODUCTION : This study describes the occurrence of trypanosomatids in phlebotomines in Brasília, Brazil. METHODS : Two hundred and ten females of 13 sand fly species were analyzed by polymerase chain reaction (PCR) using different molecular markers (D7 24Sα rRNA, kDNA, and ITS1) and sequencing. RESULTS : PCR revealed trypanosomatid-positive samples from Nyssomyia whitmani and Evandromyia evandroi, which were negative by kDNA and ITS1 Leishmania-specific PCRs. DNA sequence analysis of D7 24Sα rRNA amplicons indicated the occurrence of Blastocrithidia sp. and Trypanosoma sp. in Nyssomyia whitmani and Evandromyia evandroi, respectively. CONCLUSIONS : Two trypanosomatid species other than Leishmania sp. were found to circulate in sand flies in Central Brazil.

Conventional approach x laparoscopic abdominoperineal resection for rectal cancer treatment after neoadjuvant chemoradiation: results of a prospective randomized trial

OBJECTIVE: The aims of this study were to evaluate the safety and efficacy of laparoscopic abdominoperineal resection compared to conventional approach for surgical treatment of patients with distal rectal cancer presenting with incomplete response after chemoradiation. METHOD: Twenty eight patients with distal rectal adenocarcinoma were randomized to undergo surgical treatment by laparoscopic abdominoperineal resection or conventional approach and evaluated prospectively. Thirteen underwent laparoscopic abdominoperineal resection and 15 conventional approach. RESULTS: There was no significant difference (p<0,05) between the two studied groups regarding: gender, age, body mass index, patients with previous abdominal surgeries, intra and post operative complications, need for blood transfusion, hospital stay after surgery, length of resected segment and pathological staging. Mean operation time was 228 minutes for the laparoscopic abdominoperineal resection versus 284 minutes for the conventional approach (p=0.04). Mean anesthesia duration was shorter (p=0.03) for laparoscopic abdominoperineal resection when compared to conventional approach : 304 and 362 minutes, respectively. There was no need for conversion to open approach in this series. After a mean follow-up of 47.2 months and with the exclusion of two patients in the conventional abdominoperineal resection who presented with unsuspected synchronic metastasis during surgery, local recurrence was observed in two patients in the conventional group and in none in the laparoscopic group. CONCLUSIONS: We conclude that laparoscopic abdominoperineal resection is feasible, similar to conventional approach concerning surgery duration, intra operative morbidity, blood requirements and post operative morbidity. Larger number of cases and an extended follow-up are required to adequate evaluation of oncological results for patients undergoing laparoscopic abdominoperineal resection after chemoradiation for radical treatment of distal rectal cancer.

Atividade farmacológica dos extratos da casca do caule do tropeiro. O Connarus fulvus, Planch

O Cannarus fulvus Planch é utilizado em Goiás por ser "bom para o coração". Este trabalho estudou, a atividade farmacológica do extrato bruto etanólico (EE) do pó da casca do caule e da fração hidrossolúvel (FH) obtida após partição benzina/água. O EE e a FH produziram em ratos e camundongos diminuição da motilidade, sonolência sem hipnose, dificuldade respiratória, analgésia e arrastamento do trem posterior; estes proporcionais às doses. As DL50 em camundongos foram 210+/-22 e 310+/-52mg/kg, i.p. para o EE e FH respectivamente. A FH prolongou o sono barbitúrico, o tempo de reação ao calor na placa, quente e antagonizou (80%) as ações convulsivantes do pentilenotetrazol, mas não as da estricnina. Protegeu (100%) camundongos contra a ação letal do pancurônio e potencializou a ação da succinilcolina. Em preparações isolada, a FH potencializou (20%) a contração do diafragma em resposta ao estímulo elétrico do nervo frênico de ratos e reverteu o bloqueio neuromuscular produzido pela d-tubocurarina, em átrios de ratos e cobaias a freqüência e a força de contração não foram alteradas. As injeções do EE e FH em ratos anestesiados produziram hipotensão (5 a 100mg/kg); doses maiores foram letais. Os dados obtidos não confirmaram ação cardiativa direta. A planta apresentou ações depressões do S.N.C., analgésica, anticonvulsivante e descurarizante.

Synthesis and antibiotic activity of some β-Carboline alkaloids

Nine β-carboline alkaloids were synthetized and screened for antibiotic activity. Six of the compounds testes showed inhibitory activity against one or more of the microorganisms assayed.

Reação de dez clones de Seringueira (Hevea benthamiana) a três isolados de Microcyclus ulei

Avaliou-se a reação de dez clones de seringueira (Hevea benthamiana Muell. Arg.) frente a três isolados de Microcyclus ulei (P. Henn.) v. Arx, pertencentes aos grupos de patogenicidade I e II, quanto aos parâmetros monocíclicos: período de incubação, período latente, diâmetro da lesão e tipo de reação. Inoculou-se uma suspensão de 2 χ 105 conídios/ml na superfície abaxial de folíolos nos estádios B1/B2, deixou-se por 24 horas em câmara úmida a 24 ± lºC e transferiu-se para câmara de crescimento à mesma temperatura até a última avaliação aos 15 dias. A maioria dos clones testados apresentou resistência aos três isolados. Verificou-se ausência de interação entre clones e isolados com relação ao período de incubação, mas verificou-se interação altamente significativa entre clones e isolados em relação ao diâmetro de lesão. Os clones CNSAM 8218 e CNSAM 8219 foram altamente resistentes e apresentaram os menores diâmetros médios de lesão quando confrontados com o isolado EB1, diferindo significativamente dos demais. CNSAM 8212 também apresentou menor diâmetro de lesão frente ao isolado EB2. O CNSAM 8205, por sua vez, apresentou o maior diâmetro médio de lesão quando inoculado com o isolado EB1 e reação semelhante ao CNSAM 8204 quando inoculado com os isolados EB2 e MB1 e ao CNSAM 8201 quando inoculado com o isolado EB2. Concluiu-se que os clones avaliados apresentaram resistência vertical, o que os torna impróprios para um programa de melhoramento genético de seringueira que vise a obtenção de materiais com resistência horizontal ao M. ulei.

Plantas invasoras em sistemas agroflorestais com cupuaçuzeiro no município de Presidente Figueiredo (Amazonas, Brasil)

A infestação crescente de plantas invasoras nos sistemas agrícolas causa prejuízos às lavouras, com decréscimos acentuados da produtividade, quer pela competição direta pelos fatores de produção, quer pelos compostos alelopáticos liberados. Este trabalho consistiu de um levantamento e análise fitossociológica de alguns aspectos de espécies de invasoras que ocorrem em sistemas agroflorestais com cupuaçuzeiro, em três arranjos de culturas (mandioca+fruteiras; anuais+fruteiras; maracujá+fruteiras), sob três sistemas de adubação (NPK+MO, adubação com Fósforo e Fósforo+leguminosa). As coletas das plantas invasoras feitas em seis amostras de 0,25 m² por parcela foram posteriormente levadas ao laboratório para identificação. As 55 espécies identificadas estavam distribuídas em 23 famílias botânicas, sendo 43 espécies de dicotiledôneas (78,2%), 11 de monocotiledôneas (20,0%) e uma de pteridófita (1,8%). As famílias Poaceae (monocotiledônea) e Asteraceae (dicotiledônea) foram as mais freqüentes e com maior número de indivíduos. As espécies mais freqüentes e com maior número de plantas por m² foram Paspalum conjugatum P.J. Bergius (área A) e Homolepis aturensis (Kunth) Chase (área B), ambas da família Poaceae; Ageratum conyzoides L. da família Asteraceae, apresentou a maior densidade. Os coeficientes de similaridade variaram entre as áreas amostradas, sendo os maiores índices observados nos tratamentos que receberam adubação com matéria orgânica, particularmente nos sistemas mandioca+fruteiras. As práticas agrícolas e os sistemas de manejo do solo e das lavouras exerceram influência na composição florística e no tamanho das comunidades de plantas invasoras em cada local. O número de monocotiledôneas foi menor no tratamento com adubação NPK+MO.