Serological and Molecular Study of the Duffy Blood Group among Malarial Endemic Region Residents in Brazil

ABSTRACT Background: The atypical chemokine receptor 1 (ACKR1) gene encodes the Duffy blood group antigens in two allelic forms: FY*A (FY*01) and FY*B (FY*02), which define the Fy(a+b-), Fy(a-b+), and Fy(a+b+) phenotypes. FY*BES (FY*02N.01) is a single T to C substitution at nucleotide -67 that prevents the FY*B from being expressed in red blood cells (RBCs). Methods: We evaluated 250 residents from a Brazilian malarial endemic region (RsMR). All individuals were phenotyped for Fya and Fyb antigens and genotyped for FY*A, FY*B, FY*B SE , and FY*B weak alleles. Results: Among the 250 individuals, 209 (83.6%) reported previous malaria infection, and 41 (16.4%) did not. The Fy(a+b+) phenotype was present in 97/250 (38.8%), while the Fy(a-b-) was present in 7/250 (2.8%). The FY*A/FY*B was found in 130/250 (52%) and the FY*A/FY*A in 45/250 (18%). The c.1-67>TC was present, in homozygosity, in 11/250 (4.4%). Among 34 individuals with the Fy(a+b-) and FYA*/FYB* mutations, 4/34 (11.8%) had homozygosity for the c.1-67T>C. One individual presented the Fy(a+b-), FY*A/FY*B, and c.1-67T>C in homozygosis, whereas the other presented the Fy(a+b-), FY*A/FY*A, and c.1-67T>C in heterozygosis. Conclusions: We reported a low prevalence of the Fy(a-b-) in persons who had previously been infected with Plasmodium vivax (67.5%). We observed that 102/141 (72.3%) individuals expressing the Fyb antigen had a P. vivax infection, indicating the importance of the Fyb antigen, silenced by a c.1-67T>C mutation in homozygosis, in preventing the P. vivax infection. We showed that the c.1-67T>C mutation in the FY*A did not silence the FY*A expression on RBCs.

Efeito de bioterápico na eosinofilia durante a SLMV experimental / Biotherapic effect in eosinophilia during experimental VLMS

O Toxocara canis (Tc) é um parasito pertencente ao filo Nematódeo que possui como hospedeiro definitivo os cães, O homem é hospedeiro paratênico e contamina-se acidentalmente ao ingerir ovos contendo larvas infectantes (L3) do parasito, as quais são liberadas e atravessam a mucosa intestinal, atingem a circulação, Durante este processo migratório, antígenos de excreção e secreção (TES) são liberados provocando intensa reação inflamatória, do tipo Th2, caracterizando a síndrome, denominada Larva Migrans Visceral (SLMV), As principais características desta doença crônica são as eosinofilias sanguínea e tecidual persistentes, Desse modo, torna-se importante a busca por terapias que contribuam com a redução dos quadros inflamatórios com intensa eosinofilia, Assim, o uso deste bioterápico, produzido a partir do extrato antigênico de ovos e larvas de (Tc), e seu efeito no recrutamento de leucócitos totais, células mononucleares e eosinófilos no sangue, para o espaço broncoalveolar e para a cavidade peritoneal de camundongos infectados pelo (Tc) foi investigado, Foram utilizados camundongos fêmeas (Swiss), divididos nos grupos: Controle (C), Infectado (Tc), Imunizado (Im+Tc) e Tratado (Tc+Bio), Os animais Tc, Im+Tc e Tc+Bio receberam 500 ovos/animal por gavagem, Posteriormente, os animais foram eutanasiados no 18º dia da infecção e o número das células nos compartimentos foi determinado, Os resultados obtidos demonstraram que, Im+Tc, assim como nos Tc+Bio tiveram redução significativa dessas células nos compartimentos analisados quando comparados grupo Tc, Assim, sugeriu-se que a bioterapia modulou negativamente o recrutamento de células inflamatórias, principalmente eosinófilos no sangue, pulmão e intestino demonstrando um potencial anti-inflamatório desse bioterápico na SLMV experimental...

The Toxocara canis (Tc) is a parasite that belongs to the nematode phylum and has dogs as definitive host. The men can be accidentally contaminated by ingesting eggs containing infective larvae of the parasite. These larvae, when ingested, pass through the intestinal mucosa, reach the portal circulation and migrate through different tissues of the host. During this process, excretory-secretory antigens (TES) are released causing an intense inflammatory reaction, the Th2 type, characterizing the syndrome, called Visceral Larva migrans (VLMS). The main features of this chronic disease are blood and tissue eosinophilias. Thus, it is important to search for therapies that may contribute to the reduction of inflammatory conditions with intense eosinophilia. In this study, we investigated the use of a biotherapic produced from the antigenic extract from eggs and larvae (Tc) and its effect on the recruitment of total leukocyte, mononuclear cells and eosinophils in blood, bronchoalveolar space and peritoneal cavity of mice infected with (Tc). Female mice (Swiss) were used divided in three groups: control (C), Infected (Tc) Immunized (Im + Tc) and Treaty (Tc + Bio). The animals Tc, Im + Tc and Tc + Bio received 500 eggs / animal by gavage. Subsequently, the animals were euthanized on day 18 after infection and the number of cells in the compartments was determined. Our results showed that, Im + Tc, as in Tc + Bio had reduced these cells in compartments analyzed compared to Tc group. Thus, it was suggested that the biotherapy negatively modulated the recruitment of inflammatory cells, particularly eosinophils in blood, lung and intestine demonstrating an anti-inflammatory potential of the biotherapic in the experimental VLMS...

Preparation, characterization and evaluation of the in vivo trypanocidal activity of ursolic acid-loaded solid dispersion with poloxamer 407 and sodium caprate

Ursolic acid is a promising candidate for treatment of Chagas disease; however it has low aqueous solubility and intestinal absorption, which are both limiting factors for bioavailability. Among the strategies to enhance the solubility and dissolution of lipophilic drugs, solid dispersions are growing in popularity. In this study, we employed a mixture of the surfactants poloxamer 407 with sodium caprate to produce a solid dispersion containing ursolic acid aimed at enhancing both drug dissolution and in vivo trypanocidal activity. Compared to the physical mixture, the solid dispersion presented higher bulk density and smaller particle size. Fourier Transform Infrared Spectroscopy results showed hydrogen bonding intermolecular interactions between drug and poloxamer 407. X-ray diffractometry experiments revealed the conversion of the drug from its crystalline form to a more soluble amorphous structure. Consequently, the solubility of ursolic acid in the solid dispersion was increased and the drug dissolved in a fast and complete manner. Taken together with the oral absorption-enhancing property of sodium caprate, these results explained the increase of the in vivo trypanocidal activity of ursolic acid in solid dispersion, which also proved to be safe by cytotoxicity evaluation using the LLC-MK2 cell line.

O ácido ursólico é um candidato promissor para o tratamento da doença de Chagas, contudo este fármaco possui baixa solubilidade aquosa e limitada absorção intestinal, ambos os fatores limitantes da biodisponibilidade. Entre as estratégias para potencializar a solubilidade e a dissolução de fármacos lipofílicos, as dispersões sólidas estão crescendo em popularidade. Neste estudo, empregamos mistura dos tensoativos, poloxamer 407 e caprato de sódio, para produzir dispersão sólida contendo ácido ursólico, com o objetivo de aumentar tanto a dissolução do fármaco quanto a atividade tripanocida in vivo. Comparada à mistura física, a dispersão sólida apresentou maior densidade e menor tamanho de partícula. Os resultados da análise de espectroscopia no infravermelho com transformada de Fourier mostraram interações intermoleculares do tipo ligações de hidrogênio entre o fármaco e o poloxamer 407. Os experimentos de difratometria de raio-X revelaram a conversão do fármaco de sua forma cristalina para a forma amorfa, mais solúvel. Consequentemente, a solubilidade do ácido ursólico em dispersão sólida foi aumentada e o fármaco dissolveu-se de maneira mais rápida e completa. Em conjunto com as propriedades promotoras de absorção oral do caprato de sódio, estes resultados explicaram o aumento da atividade tripanocida in vivo do ácido ursólico em dispersão sólida, que também se provou segura após avaliação de citotoxicidade empregando a linhagem celular LLC-MK2.

Alteraciones histopatológicas de placentas de ratones infectadas con diferentes cepas de Trypanosoma cruzi / Histopathological alterations of placentas from mice infected by different Trypanosoma cruzi strains

El objetivo del presente estudio fue evaluar las alteraciones histopatológicas de las placentas provocadas por la infección materna durante la preñez con diferentes cepas de Trypanosoma cruzi. Ratones preñadas, de 60 días de edad, fueron inoculadas por la via intraperitoneal (i.p.) con 2 x 105 tripomastigotes sanguíneos de las cepas colombiana, Y, Bolivia o RC del T. cruzi. Los resultados mostraron que las cepas Bolivia y RC presentan mayor histotropismo por las células placentarias del ratón que las cepas colombiana e Y. El proceso inflamatorio afectó la decidua basal en todos los grupos de ratones infectados, mientras que la cepa colombiana causó intensa degeneración de la decidua basal, con células inflamatorias escasas o ausentes. Las placentas de todos los grupos infectados presentaron áreas de edema, degeneración, necrosis y calcificación, principalmente en la decidua basal. En los animales inoculados con la cepa RC del T. cruzi también fue afectado el trofoblasto de la zona esponjosa. El parasitismo más intenso fue observado en los animales infectados con las cepas Bolivia y RC, envolviendo todas las zonas placentarias, incluso las células trofoblásticas gigantes y el trofoblasto esponjoso. Los resultados permiten concluir que las placentas sufren procesos patológicos diferentes, característicos para cada cepa del T. cruzi.

The objective of the present study was to evaluate the histopathological alterations of the placenta provoked by maternal infection during pregnancy by different Trypanosoma cruzi strains. Pregnant mice, 60 day-old, were i.p. inoculated with 2 x 105 bloodstream trypomastigotes of colombian, Y, Bolivia or RC strains of T. cruzi. The results demonstrated that the Bolivia and RC strains showed higher histotropism for the mouse placental cells than the colombian or Y strains. The inflammatory process in all the infected groups involved the basal decidua, but the colombian strain caused intense decidual degeneration, with rare or absent inflammatory cells. Placentas from all the infected groups displayed aéreas of edema, degeneration, necrosis and calcification, mainly in the basal decidua, but the group infected by the RC strain showed also involvement of the spongiotrophoblast. The highest parasitism was observed with the Bolivia and RC strains, involving all the placenta zones, including the trophoblast giant cells and the spongiotrophoblast. The results show that placentas undergo differentiated pathological processes, characteristic for each strain.

Trypanocidal activity of Brazilian plants against epimastigote forms from Y and Bolivia strains of Trypanosoma cruzi

Chagas disease is one of the main public health problems in Latin America. Since the available treatments for this disease are not effective in providing cure, the screening of potential antiprotozoal agents is essential, mainly of those obtained from natural sources. This study aimed to provide an evaluation of the trypanocidal activity of 92 ethanol extracts from species belonging to the families Annonaceae, Apiaceae, Cucurbitaceae, Lamiaceae, Lauraceae, Moraceae, Nyctaginaceae, and Verbenaceae against the Y and Bolivia strains of Trypanosoma cruzi. Additionally, cytotoxic activity on LLCMK2 fibroblasts was evaluated. Both the trypanocidal activity and cytotoxicity were evaluated using the MTT method, in the following concentrations: 500, 350, 250, and 100 µg/mL. Benznidazole was used for positive control. The best results among the 92 samples evaluated were obtained with ethanol extracts of Ocotea paranapiacabensis (Am93) and Aegiphila lhotzkiana (Am160). Am93 showed trypanocidal activity against epimastigote forms of the Bolivia strain and was moderately toxic to LLCMK2 cells, its Selectivity Index (SI) being 14.56, while Am160 showed moderate trypanocidal activity against the Bolivia strain and moderate toxicicity, its SI being equal to 1.15. The screening of Brazilian plants has indicated the potential effect of ethanol extracts obtained from Ocotea paranapiacabensis and Aegiphila lhotzkiana against Chagas disease.

Association between Knops blood group polymorphisms and susceptibility to malaria in an endemic area of the Brazilian Amazon

Complement receptor 1 (CR1) gene polymorphisms that are associated with Knops blood group antigens may influence the binding of Plasmodium parasites to erythrocytes, thereby affecting susceptibility to malaria. The aim of this study was to evaluate the genotype and allele and haplotype frequencies of single-nucleotide polymorphisms (SNPs) of Knops blood group antigens and examine their association with susceptibility to malaria in an endemic area of Brazil. One hundred and twenty-six individuals from the Brazilian Amazon were studied. The CR1-genomic fragment was amplified by PCR and six SNPs and haplotypes were identified after DNA sequence analysis. Allele and haplotype frequencies revealed that the Kn b allele and H8 haplotype were possibly associated with susceptibility to Plasmodium falciparum. The odds ratios were reasonably high, suggesting a potentially important association between two Knops blood antigens (Kn b and KAM+) that confer susceptibility to P. falciparum in individuals from the Brazilian Amazon.

Trypanocidal activity of flavonoids and limonoids isolated from Myrsinaceae and Meliaceae active plant extracts / Atividade tripanocida de flavonoides e limonoides isolados de extratos ativos de plantas de Myrsinaceae e Meliaceae

The activity of crude extracts of three Rapanea species (Myrsinaceae) and Cipadessa fruticosa (Meliaceae) was evaluated in vitro against the trypomastigote forms of Trypanosoma cruzi. Thirty-three extracts from different organs of these species were assayed and eleven of them showed significant activity (lysis percent >50). The fractionation of an active extract from branches of R. lancifolia (99.5 percent) led to the isolation of two flavonoids: quercetin and taxifolin, which have weak trypanocidal activity. Additionally, one active extract from fruits of C. fruticosa (97.7 percent) afforded mexicanolide limonoids: cipadesin, mexicanolide, febrifugin and cipadesin A, that were slightly active on T. cruzi. Moreover, other two flavonoids (flavone and 7-methoxyflavone), previously assayed against T. cruzi, were isolated from the hexane extract from branches of C. fruticosa (100 percent). The results presented here suggest that the plants evaluated could be a source of new active compounds against T. cruzi.

A atividade de extratos brutos de três espécies de Rapanea (Myrsinaceae) e de Cipadessa fruticosa (Meliaceae) foi avaliada in vitro contra formas tripomastigotas de Trypanosoma cruzi. Foram obtidos 33 extratos de diferentes órgãos das espécies estudadas, sendo que onze deles apresentaram atividades significantes ( por cento de lise > 50) nos ensaios realizados. O fracionamento de um extrato ativo dos galhos de R. lancifolia (99,5 por cento) resultou no isolamento de dois flavonoides (quercetina e taxifolina), que apresentaram baixa atividade tripanocida. De um extrato ativo dos frutos de C. fruticosa (97,7 por cento) foram isolados os limonoides mexicanolídeos cipadesina, mexicanolídeo, febrifugina e cipadesina A, que foram moderadamente ativos sobre T. cruzi. Além disso, outros dois flavonoides (flavona e 7-metoxiflavona), previamente ensaiados contra T. cruzi, foram isolados do extrato hexânico dos galhos de C. fruticosa (100 por cento). Os resultados obtidos aqui sugerem que as plantas avaliadas podem constituir fontes de novas substâncias ativas sobre o T. cruzi.

Arrabidaea chica (HBK) Verlot: phytochemical approach, antifungal and trypanocidal activities / Arrabidaea chica (HBK) Verlot: abordagem fitoquímica, atividades tripanocida e antifúngica

Arrabidaea chica (HBK.) Verlot (Bignoniaceae) vernacular name "Pariri", is a climbing shrub, widespread from South Mexico to Guyana and central Brazil and is traditionally indicated to treat symptoms of inflammations and skin affections. Its ethanol extract was chemically investigated and tested against yeasts and dermatophytic fungi. The trypanocidal activity of the same extract was also evaluated. This work reports the isolation of three flavonoids, the total growth inhibition of Trichophyton mentagrophytes and a significant trypanocidal effect of the ethanol extract and its fractions. No relevant acute toxicity was detected even at a dose of 1000 mg/kg.

Arrabidaea chica (HBK.) Verlot (Bignoniaceae) popularmente, "Pariri", é um arbusto escandente, distribuído do sul do México até a Guiana e Brasil central e é tradicionalmente indicado para tratar sintomas de inflamações e afecções da pele. Seu extrato etanólico foi quimicamente investigado e testado contra leveduras e fungos dermatófitos. A atividade tripanocida do mesmo extrato foi também avaliada. Este trabalho reporta o isolamento de três flavonóides, a inibição total do crescimento de Trichophyton mentagrophytes e um significante efeito tripanocida do extrato etanólico e de suas frações. Não foi detectada qualquer toxicidade aguda relevante, mesmo a uma dose de 1000 mg/kg.

Estudio cariométrico de placentas de ratones con infección aguda por diferentes cepas de Trypanosoma cruzi / Karyometric study of placentas from mice with acute infection by different strains of Trypanosoma cruzi

El objetivo de este trabajo fue evaluar cariometricamente las alteraciones causadas por diferentes cepas de T. cruzi en la placenta del ratón. Ratones hembras de 60 días, grávidas, fueron inoculadas, intraperitonealmente, con 2 x 10(5) tripomastigotes sanguíneos de las cepas colombiana, Y, Solivia o RC del T. cruzi. Fueron observadas claras diferencias en las alteraciones cariométricas de las células trofoblásticas gigantes y de las células trofoblásticas de la zona esponjosa. Los resultados demostraron que las cepas colombiana y RC causan alteraciones tanto en las células trofoblásticas gigantes como en las células del trofoblasto esponjoso, mientras que las cepas Y y Bolivia provocan alteraciones solamente en las células trofoblásticas gigantes. Es posible concluir que cada cepa posee características propias y que, a pesar del tipo similar de transmisión, presenta matices diferenciales en el proceso de la patogénesis placentaria.

The objective of this work was to evaluate karyometrically the alterations caused by different strains of Trypanosoma cruzi in the mouse placenta. Pregnant mice, 60-day old, were intraperitoneally inoculated with 2 x 10(5) bloodstream trypomastigotes of the Colombian, Y, Bolivia or RC strain of T cruzi. There were observed clear differences in the karyometric alterations of the trophoblast giant cells and in the spongiotrophoblast cells. The results demonstrate that the Colombian and RC strains cause alterations both in the trophoblast giant cells and in the spongiotrophoblast cells, whereas the Y and Bolivia strains provoke alterations only in the trophoblast giant cells. It is possible concluding that each strain has its own characteristics and that, in spite of the similar type of transmission, it show differential nuances in the placental pathogenic process.

Histometría de la glándula sublingual de ratones (Mus musculus) machos y hembras infectados con la cepa RAL del parásito de Chagas, Trypanosoma cruzi

Histometry of the sublingual gland in male and female mice (Mus musculus) infected with the RAL strain of the Chagas parasite, Trypanosoma cruzi. The aim of this work was to analyze histologically and histometrically the sublingual gland of mice infected with the RAL strain of T. cruzi, according to the sex. Swiss mice (Mus musculus) were inoculated with 2x104 blood trypomastigotes of the RAL strain of T. cruzi. in the peak of the parasitemia (12th day) the mice were sacrificed, and the sublingual glands were fixed in ALFAC. HE-stained histological sections were evaluated histometrically. The parasitemia was higher in females. Histopatologically, acini of the infected animals were smaller, with scanty production of secretion, and smaller striated ducts. The nuclei of the demilunes were smaller and showed amastigote nests in the cytoplasm. Karyometrically, nuclei of the acini, demilunes and striated ducts were smaller in the infected mice. Stereologically, it was observed that relative volumes of acini and ducts were smaller and, inversely, relative volumen were greater for the conjunctive tissue in the infected males. The surface densities of acini and ducts were bigger and the diameter and thickness of the wall were smaller in this group. On the other hand, relative volume of acini was smaller and those of the ducts and conjunctive tissue were bigger in the infected females. The diameter and thickness of the wall of acini were smaller, and those of the striated ducts were bigger in this group. The RAL strain of T. cruzi caused general atrophy in the sublingual gland, with numerous nests of parasites in the glandular parenchyma. Rev. Biol. Trop. 56 (2): 459-471. Epub 2008 June 30.

Analizamos morfológica e histométricamente la glándula sublingual de ratones infectados con la cepa RAL del Trypanosoma cruzi, en machos y hembras. Usamos ratones albinos (Mus musculus), variedad Swiss, inoculados con 2x104 tripomastigotes sanguíneos de la cepa RAL del T. cruzi.. Sacrificamos los animales en el pico de la parasitemia (12º día) y fijamos las glándulas sublinguales en ALFAC. Los cortes histológicos teñidos con HE fueron evaluados histométricamente (cariometría y estereología). La parasitemia fue más elevada en las hembras. Histopatológicamente, los "ácinos" (acini) de los animales infectados eran menores, con escasa secreción, y conductos estriados menores. Los núcleos de las "medialunas" eran menores y había nidos de amastigotes en el citoplasma. Cariométricamente, los núcleos de los ácinos, medialunas y conductos estriados eran menores en los ratones infectados. Estereológicamente, los volúmenes relativos ocupados por ácinos y conductos estriados fueron menores e, inversamente, fue mayor el volumen para el tejido conjuntivo de los machos infectados. Las densidades de superficie de ácinos y conductos fueron mayores, y el diámetro y el espesor de la pared menores, en este grupo. Por otro lado, la densidad de ácinos fue menor, y las de los conductos estriados y tejido conjuntivo, fueron mayores en las hembras infectadas. Las densidades de superficie de ácinos y conductos estriados fueron mayores, mientras que el diámetro y espesor de la pared de los ácinos fueron menores (y las de los conductos estriados mayores), en este grupo. La cepa RAL del T. cruzi causó un cuadro general de atrofia general en la glándula sublingual, con numerosos nidos de parásitos.

Intraoperative topical administration of mitomycin C, in different concentrations, on the cicatrization of mioplasties of the dorsal rectus of rabbits / Administração tópica per-operatória da mitomicina C, em diferentes concentrações, sobre a cicatrização de mioplastias do reto dorsal de coelhos

Myoplasties of the extraocular muscle may cause adhesions between the operated muscle and the adjacent tissues, commonly generating cicatricial strabismus. With the purpose of reducing to a minimum the occurrence of adhesion, the effects of mitomycin C, an antifibrotic agent, were studied in concentrations of 0.008 percent, 0.02 percent, and 0.04 percent applied during intraoperative of myoplasties of the superior rectus muscle of rabbits. Fifty six animals were divided in five groups. During the postoperative, the operated areas were washed with physiological solution. Eye drop instillation to prevent inflammation and bacterial infection were used. The method to analyze the results consisted of clinical and histological evaluation and statistical analyzes. We also evaluated at the same time the amount of basic fibroblast growth factor (FGF-2) by immunohistochemical study. Clinically, more adhesions were found in the eyes of the control group than in the groups of treated eyes. However there was no significant statistics difference between the two groups (P>0.05). Histologically, mitomycin C caused a delayed cicatrization in the mioplastic areas, specially in the group who received the 0.02 percent concentration. The immunohistochemical showed FGF-2 marking in fibroblasts and macrophages, but between the groups there wasn't no difference. Based on those results, mitomycin C in the utilized concentrations was capable of delaying the cicatrization and consequently avoid the secondary strabismus without undesirable side effects.

Mioplastias da musculatura extra-ocular podem ensejar aderências entre o músculo operado e os tecidos adjacentes, produzindo, não raro, estrabismos cicatriciais. Com intuito de se minimizar a ocorrência de aderências, investigaram-se os efeitos da mitomicina C, como agente antifibrótico, em concentrações ascendentes de 0,008, 0,02 e 0,04 por cento, aplicada no per-operatório de mioplastias do reto superior do bulbo do olho de coelhos. Operaram-se 56 animais, que compuseram cinco grupos. No pós-operatório, instituíram-se limpeza com solução fisiológica das áreas operadas e profilaxia antimicrobiana e antiinflamatória, na forma de colírio. Avaliações clínica, histológica e imunoistoquímica, em que se estudou o fator de crescimento fibroblástico-básico (FGF-2), e estatística compuseram os métodos de análise dos resultados. Encontraram-se, clinicamente, mais aderências nos olhos-controle, comparativamente aos tratados, embora sem diferença estatística (P>0,05). À histologia, verificou-se que a mitomicina C ensejou retardo da cicatrização junto às áreas das mioplastias, notadamente no grupo que a recebeu, à concentração de 0,02 por cento. A imunoistoquímica revelou marcação do FGF-2 em fibroblastos e macrófagos indistintamente entre os grupos. Com base nos resultados, permite-se admitir que a mitomicina C, nas concentrações em que foi empregada, retardou a cicatrização e, por conseguinte, o estrabismo secundário, sem ensejar efeitos colaterais.

Morfometría de la parótida de ratón infectado por la cepa RAL de Trypanosoma cruzi en función del sexo / Morphometry of the parotid Gland in mice infected by the RAL strain of Trypanosoma cruzi in function of the sex

Fueron estudiadas las alteraciones y el nivel de deterioro de las parótidas de animales infectados con la cepa RAL de Trypanosoma cruzi. Se utilizaron ratones albinos (Mus musculus) de ambos sexos, debido al dimorfismo sexual de las glándulas salivares, inoculados con 2 x 104 tripomastigotes sanguíneos de la cepa RAL del T. cruzi. Los animales fueron sacrificados al 12° día de infección, coincidiendo con el pico parasitémico. La parótida fue procesada histológicamente y, posteriormente, evaluada histopatológica y morfométricamente. Los resultados permitieron verificar intenso parasitismo en la glándula, la que presentó desorganización estructural y atrofia generalizada de acinos y conductos, más intensos en las hembras. Concluyendo, la cepa RAL del T. cruzi mostró un comportamiento atípico en relación a otras cepas, causando modificaciones más evidentes en las hembras, debido, posiblemente, a alteraciones hormonales desencadenadas por el T. cruzi.

They were studied the alterations and the level of deterioration of the parotid gland in mice infected with the RAL strain oí Trypanosoma cruzi. They were used albino mice (Mus musculus) of both sexes, due to the existence of sexual dimorphism of the salivary glands, inoculated with 2 x 104 blood trypomastigotes of the RAL strain of T cruzi. The animals were sacrificed at the 12th day of infection, coinciding with the parasitemic peak, and the parotid gland was histologically processed and histopathologically and histometrically studied. The results allow verifying intense parasitism in the parotid gland with structural disorganization and widespread atrophy of acini and ducts, more marked in the females. Concluding, the RAL strain of T cruzi shows an atypical behavior in relation to other strains, provoking more clear modifications in the females, probably due to the hormonal alterations motivated by the T cruzi.

Effect of extracts and isolated compounds from Chresta scapigera on viability of Leishmania amazonensis and Trypanosoma cruzi

Fractionation of bioactive crude extracts of Chresta scapigera led to the isolation of four triterpenes and five flavonoids, among them beta-amyrin acetate (1), 11alpha,12alpha-oxidetaraxeryl acetate (2) and lupeol (3), as well as the flavonoids apigenin (6), kaempferol (7), crysoeriol (8) and luteolin (9) were active against Leishmania amazonensis amastigotes-like stages, while only the flavonoids (6), (7) and (9) showed toxicity towards bloods trypomastigote forms of Trypanosoma cruzi.

O fracionamento dos extratos bioativos de Chresta scapigera proporcionou o isolamento de triterpenos e flavonóides, dentre os quais acetato de beta-amirina (1), acetate de 11alfa,12-oxidotaraxeril (2) e lupeol (3), assim como os flavonóides apigenina (6), caenferol (7), crisoeriol (8) e luteolina (9) mostraram-se ativos contra formas amastigotas de Leishmania amazonensis, enquanto, apenas os flavonóides (6), (7) e (9) apresentaram toxicidade contra as formas tripomastigotas de Trypanosoma cruzi.

Estudio morfométrico del hígado de ratón en la enfermedad de Chagas experimental / Morphometrical study of the mouse liver in the experimental Chagas' disease

El objetivo de este trabajo fue caracterizar histopatológicamente y morfométricamente las alteraciones del tejido hepático de ratón, durante la fase aguda de la infección por la cepa MORC-2 de Trypanosoma cruzi. Esta cepa mostró acentuado tropismo por el hígado, con numerosos nidos de amastigotes en los cortes examinados. El hígado de los animales infectados estaba constituido por células menores, con citoplasma granuloso. En algunas áreas, los sinusoides estaban congestionados y las células de Kupffer hipertróficas e hiperplásicas. El tejido hepático mostró focos circunscritos de células inflamatorias en áreas de necrosis, sinusoides, en torno de las venas centrolobulillares y de los espacios porta. La vena centrolobulillar estaba dilatada y congestionada, con necrosis focales y ruptura de la pared en algunos campos. Los espacios porta estaban desorganizados, a veces, con intenso infiltrado inflamatorio. En algunas áreas fue posible observar degeneración cística (spongis hepatis). Por todo el tejido hepático se observaron nidos de amastigotes, de tamaño variable, algunos rodeados por infiltrado inflamatorio crónico. En el espacio porta, el volumen relativo de los conductos biliares y vasos sanguíneos, así como la densidad de superficie de las arterias fueron mayores en el grupo infectado.

The objective of this work was to characterize histopatologically and morphometrically the alterations of the mouse liver during the acute infection by the MORC-2 strain of Trypanosoma cruzi. This strain showed marked tropism by the liver, with numerous nests of amastigotes in the examined sections. The liver of the infected animals was constituted by smaller cells, with granular cytoplasm. In some areas, the sinusoids were congested and the Kuppfer cells were hipertrofied and hiperplasic. The hepatic tissue showed circumscribed foci of inflammatory cells into necrotic areas, sinusoids, around the contrilobular veins and the portal spaces. The centrilobular vein was dilated and congested, with focal necrosis and rupture of the wall in some regions. The portal spaces were disorganized, sometimes with intense inflammatory infiltrate. In some areas it was possible to observe cystic degeneration (spongis hepatis). In the hepatic tissue, nests of amastigotes, of variable sizes, were observed, some surrounded by chronic inflammatory infiltrate. In the portal space, the relative volume of the biliary ducts and blood vessels, as well as the surface density of the arteries was greater in the infected group.

Biological activities and chemical composition of crude extracts from Chresta exsucca

As atividades tripanocida, leishmanicida, antibacteriana e antifungica dos extratos brutos de Chresta exsucca foram investigadas. Formas tripomastigotas do Trypanosoma cruzi, formas amastigotas de Leishmania amazonensis e vinte cepas de microrganismos, incluindo bactérias Gram-positivas, Gram-negativas e leveduras, foram utilizadas nos ensaios biológicos. Os melhores resultados foram obtidos para a atividade leishmanicida. A composição química dos extratos hexânicos e etanólicos dessa espécie foi determinada empregando-se técnicas cromatográficas como HRGC e HPLC-ESI-MS, respectivamente. Esteróides, triterpenos e flavonóides foram identificados.

Trypanocidal activity of Meliaceae and Rutaceae plant extracts

The in vitro trypanocidal activity of 22 extracts and 43 fractions of plants belonging to the families Meliaceae and Rutaceae was evaluated. The extracts from leaves of Conchocarphus heterophyllus and branches of Trichilia ramalhoi were the most active. The trypanocidal activity seems to be increased by fractionation of the extracts. Fractions from C. heterophyllus and Galipea carinata were the most active and a 100 percent lysis of the parasites was observed for five fractions. From one of them were isolated two flavonoids: flavone and 7-methoxyflavone, which showed weak trypanocidal activity. The results obtained from the extracts and fractions revealed that the order Rutales is a promising source for the search of new drugs for Chagas disease. Phytochemical studies with the other active fractions are underway in order to isolate compounds, which could be associated with observed activities.

Avaliação da atividade biológica do bioterápico in vivo sob as formas amastigotas na infecção experimental determinada por Leishmania (Viannia) braziliensis / Evaluation of the biological activity

As infecções por agentes do gênero Leishmania constituem um importante problema de saúde pública na Américas, sendo o Brasil o país que apresenta a mais alta incidência. Transmitida por fêmeas do inseto do gênero Lutzomyia, a Leishmania (Viannia) braziliensis, agente etiológico da Leishmaniose tegumentar americana (LTA), é uma zoonose, que impõe como principal característica patogênica uma infecção que varia de casos benignos, apresentando-se como lesão localizada, a casos que chegam a atingir tecidos mais profundos. O tratamento para essa doença é altamente tóxico, provocando diversos efeitos colaterais. Com base nesse fato, o objetivo deste trabalho foi avaliar o bioterápico (realizado com formas infectantes da Leishmania) como um tratamento alternativo para a Leishmaniose. Camundongos machos Mus musculus foram previamente infectados com a Leishmania (Viannia) braziliensis e depois tratados por três meses. Nos primeiros 30 dias de tratamento houve uma redução no diâmetro das lesões; no final, observou-se uma agravação dessas lesões, mas quando se comparou o grupo tratado com o controle negativo, as lesões estavam significantemente menores, mostrando um efeito terapêutico do bioterápico.

Evaluation of trypanocidal activity from Calea uniflora (Heliantheae-Asteraceae) extracts

Dichloromethane crude extract from xylopodium of Calea uniflora Less (Heliantheae-Asteraceae) showed in vitro trypanocidal activity against trypomastigote forms of Trypanosoma cruzi. The data obtained allow to conclude that the crude extract must be investigate to identify its active compounds.

Estudios morfológico y morfométrico del hígado de fetos de ratas con enfermedad de Chagas aguda / Morphologic and morphometric study of the fetus liver from mice with acute Chagas' disease

Fueron estudiados los fetos de la enfermedad de Chagas aguda, durante la preñez en fetos de ratas, particularmente las posibles alteraciones provocadas en las células hepáticas. Fue demostrado un retardo en el crecimiento intrauterino de los fetos, que presentaron peso y longitud menores que el grupo control. A nivel hepático no fue identificada la presencia de nidos de parásitos. Sin embargo, el hígado de los fetos del grupo inoculado presentaba algunos hepatocitos degenerados y con desorganización arquitectural, mientras que los capilares sinusoides estaban congestivos y dilatados. Morfométricamente se verificó la ausencia de alteraciones nucleares en los hepatocitos de los fetos del grupo inoculado. El volumen relativo ocupado por hepatocitos, fue significativamente mayor en el hígado de los fetos del grupo inoculado que en el grupo control, al contrario de lo observado con el volumen relativo de los sinusoides. Así, aún dilatados y congestionados, el volumen relativo de los sinusoides hepáticos en los fetos del grupo inoculado, fue menor que en los controles, reflejando una disminución del número de capilares sinusoides