Biologic and Genetics Aspects of Chagas Disease at Endemic Areas

The etiologic agent of Chagas Disease is the Trypanosoma cruzi, transmitted through blood-sucking insect vectors of the Triatominae subfamily, representing one of the most serious public health concerns in Latin America. There are geographic variations in the prevalence of clinical forms and morbidity of Chagas disease, likely due to genetic variation of the T. cruzi and the host genetic and environmental features. Increasing evidence has supported that inflammatory cytokines and chemokines are responsible for the generation of the inflammatory infiltrate and tissue damage. Moreover, genetic polymorphisms, protein expression levels, and genomic imbalances are associated with disease progression. This paper discusses these key aspects. Large surveys were carried out in Brazil and served as baseline for definition of the control measures adopted. However, Chagas disease is still active, and aspects such as host-parasite interactions, genetic mechanisms of cellular interaction, genetic variability, and tropism need further investigations in the attempt to eradicate the disease

Microdissection testicular sperm extraction causes spermatogenic alterations in the contralateral testis

Testicular sperm extraction (TESE) associated with intracytoplasmic sperm injection has allowed many men presenting non-obstructive azoospermia to achieve fatherhood. Microdissection TESE (microTESE) was proposed as a method to improve sperm retrieval rates in these patients; however, there have been failures. Little is known about whether microTESE leads to spermatogenic alterations in the contralateral testis. We assessed histological outcomes of experimental microTESE in the contralateral testis of adult male rabbits. Nine adult male rabbits were divided into three groups: control (testicular biopsy to observe normal histological and morphometric values), sham (incision of the tunica vaginalis, and a contralateral testicular biopsy to observe histological and morphometric patterns, 45 days later), and study (left testicular microTESE, and a right testicular biopsy to observe histological and morphometric patterns, 45 days later). Sections were assessed by calculating Johnsen-like scores, and measuring total tubule diameter, lumen diameter and epithelial height. the results were compared using ANOVA and Bonferroni's statistical analysis. Morphometric evaluation of the seminiferous tubules did not demonstrate differences between the three groups. However, microTESE caused spermatogenic alterations, leading to maturation arrest in the contralateral testis.Universidade Federal de São Paulo, Dept Cirurgia, Div Urol, Secao Reprod Humana, São Paulo, BrazilUniv Estadual Paulista, Dept Biol, Inst Biociencias Letras & Ciencias Exatas Sao Jos, Sao Jose Do Rio Preto, SP, BrazilUniversidade Federal de São Paulo, Dept Cirurgia, Div Urol, Secao Reprod Humana, São Paulo, BrazilWeb of Scienc

Bright nanoparticles for an even brighter future: efficient production of luminescent carbon nanodots from olive mill wastewater

Este trabalho foi financiado pelo Concurso Anual para Projetos de Investigação, Desenvolvimento, Inovação e Criação Artística (IDI&CA) 2016 do Instituto Politécnico de Lisboa. Código de referência IPL/2016/NANOLIVE/ISELCarbon nanodots (CNDs) are a very recent class of spherical-shaped nanosized carbon materials possessing average typical diameters < 10 nm. Since the very first reports on carbon dots,1,2 a variety of methods (top-down and bottom-up strategies), carbon sources and passivating agents, have dealt with their synthesis.3 The bottom-up approach, encompassing the use of pyrolytic/solvothermal processes, is more amenable for large-scale production and can cope with a large diversity of carbon precursors, either from natural or synthetic sources, typically endowed with acid, alcohol and amine functionalities.4 Some of the interesting CNDs properties include tunable photoluminescence, outstanding photostability and negligible cytotoxicity. These unique properties have prompted their intense and widespread use in several fields, such as fluorescent bioimaging and nanomedicine, chemo/biosensing, photocatalysis and optoelectronics.4info:eu-repo/semantics/publishedVersio

Genomics and proteomics approaches to the study of cancer-stroma interactions

<p>Abstract</p> <p>Background</p> <p>The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression.</p> <p>Methods</p> <p>The study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells.</p> <p>Results</p> <p>We observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (<it>ARID4A</it>, <it>CALR</it>, <it>GNB2L1</it>, <it>RNF10</it>, <it>SQSTM1</it>, <it>USP9X</it>) were validated by real time PCR.</p> <p>Conclusions</p> <p>A significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.</p

Chagas disease and triatomine biology (Heteroptera, Triatominae), with emphasis on aspects of spermatogenesis

A century after the discovery of Chagas disease, it is still one of the most important parasitic diseases affecting humans. The subfamily Triatominae is important in medical health, because these insects are vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease. These insects are also of important cytological relevance because they have particular cell characteristics, such as persistence of nucleolar material in spermatogenesis. The germ cells of the animal kingdom have chromatoid bodies (CBs) in their cytoplasm that can originate from nucleolar material that is fragmented in the early stages of spermatogenesis and plays an important role in cellular communication between the spermatids during spermiogenesis. Currently, there are few studies on the function and formation of the CB in nucleologenesis, especially with emphasis on the ultrastructure of the cells involved in spermatogenesis of insects. Considering the importance of knowledge about the triatomine fauna, we conducted a study of the biogeography and reports of these insects and a survey of patients with Chagas disease in the northwestern region of São Paulo State. Data collected from 1995 to 2009 indicated 700 individuals with Chagas disease, demonstrating a range of 0 to 40 years, which shows that the disease may be active in this region. Moreover, of the 1150 patients treated for cardiomyopathy, 44% were chagasic. Regarding the triatomines noted and captured in the period from 2004 to 2009, the species were Triatoma sordida and Rhodnius neglectus, with T. sordida being the most abundant. In addition, some triatomines were infected by T. cruzi in various developmental stages. We also analyzed the nucleolar cycle and fibrillarin nucleolar protein expression in CB of spermatogenic cells of T. infestans and T. sordida, using histological, ultrastructural and immunocytochemical techniques. The results revealed fibrillarin nucleolar protein expression in the nucleus and in some cytoplasmic spots of germ cells during spermatogenesis in triatomines. These data suggest that fibrillarin could be a constituent of CB, which was most likely derived from nucleolar fragmentation. This is the first time that fibrillarin protein expression has been shown in CB during spermatogenesis progression in triatomines. Knowledge about the biology of triatomines was deepened in this study and, in particular, the structural and ultrastructural aspects of spermatogenesis in triatomines. This study showed that the disease may be active in the northwestern region of São Paulo and expanded our knowledge of the biology of triatomines, the main vectors of Chagas disease. © FUNPEC-RP

Study of mast cells and annexin 1 protein in laryngeal squamous cell carcinoma

O carcinoma espinocelular de laringe e uma das neoplasias mais comuns e com altas taxas de mortalidade. Trata-se de uma doenca progressiva que se origina no epitelio e invade o tecido conjuntivo, provocando alteracoes histopatologicas durante o processo de tumorigenese. No presente estudo, foram avaliadas a relacao entre os mastocitos, celulas que tem um papel bem definido na modulacao da resposta imunologica e inflamatoria, e a expressao da proteina Anexina 1 nos mastocitos do carcinoma espinocelular de laringe, nos tecidos tumoral e adjacente ao tumor. A reacao imuno-histoquimica, usando o anticorpo antitriptase, permitiu quantificar os mastocitos da laringe. Os resultados obtidos mostram que, no tecido tumoral, os mastocitos foram localizados proximos ao epitelio tumoral invasivo, infiltrados em regioes intra-epiteliais e, frequentemente, em processo de desgranulacao. Houve um aumento significativo do numero de mastocitos desgranulados no tumor em relacao ao controle. A presenca de mastocitos desgranulados triptase positivos sugere liberacao deste mediador para o tumor e o tecido adjacente. A expressao e a localizacao da proteina antiinflamatoria Anexina 1 foram monitoradas pela reacao de imunocitoquimica ultra-estrutural, utilizando-se um anticorpo especifico para a regiao N-terminal da Anexina 1. Esta foi a primeira demonstracao imunocitoquimica da presenca desta proteina em mastocitos humanos. Estas celulas sao imunorreativas para a Anexina 1 no tumor, no tecido adjacente e controle livre de neoplasias. A analise quantitativa mostrou um aumento significativo da proteina nos mastocitos do tumor e do tecido adjacente. As celulas tumorais tambem sao imunorreativas para Anexinal. Nossos resultados indicam que esta proteina e produzida e expressada em pacientes com cancer. Particularmente, os mastocitos do tecido tumoral sao ricos na expressao desta proteina em forma de agregados caracteristicos no interior dos granulos citoplasmaticos. A Anexina 1, derivada de mastocitos, pode ser um fator inibitorio no controle do...(au)BV UNIFESP: Teses e dissertaçõe

Biologia reprodutiva de Corecoris fuscus, Leptoglossus gonagra, L. zonatus e Sphictyrtus fasciatus (Heteroptera: Coreidae), com ênfase aos aspectos ultraestruturais

A Subordem Heteroptera contém cerca de 75 famílias, incluindo Coreidae, na qual pertencem as espécies Corecoris fuscus, Leptoglossus gonagra, L. zonatus e Sphictyrtus fasciatus, analisadas no presente trabalho. Os indivíduos dessa família são conhecidos por serem exclusivamente fitófagos e alimentam-se de plantas de valor comercial. Apesar da grande diversidade desta subordem, há poucos estudos relacionados com a ultraestrutura da espermátide dessas espécies. Assim, nesse trabalho, por meio da microscopia eletrônica de transmissão, foi analisado o desenvolvimento da espermiogênese na cabeça e no flagelo, com ênfase na formação do acrossomo, do adjunto do centríolo e dos derivados mitocondriais. O acrossomo tem origem a partir do complexo de Golgi, por meio da formação de uma vesícula pró-acrossomal e concomitante a esse processo é formado o adjunto do centríolo. O acrossomo determina o polo anterior do espermatozoide e o adjunto do centríolo determina o polo posterior. Observou-se também que no início do processo espermiogênico, as mitocôndrias encontram-se espalhadas no citoplasma, então gradualmente se fundem e dão origem ao complexo mitocondrial, o qual se divide, posteriormente, em duas subestruturas, denominadas derivados mitocondriais. Essas se posicionam bilateralmente ao axonema, o qual possui o padrão típico de disposição dos microtúbulos de 9 + 9 + 2. O processo de formação do acrossomo e dos derivados e a conformação dos microtúbulos do axonema são típicos de insetos. Além do padrão de microtúbulos do axonema, foram encontradas características sinapomórficas de Heteroptera, como a presença de duas estruturas paracristalinas em cada derivado mitocondrial e a presença de pontes que ligam os derivados mitocondriais ao microtúbulos axonemais, os quais são ferramentas importantes para auxiliar análises filogenéticas e taxonômicas.The suborder Heteroptera contains about 75 families, including Coreidae, to which the species Corecoris fuscus, Leptoglossus gonagra, L. zonatus and Sphictyrtus fasciatus belong. Individuals of this family are known for being exclusively phytophagous and feeding on commercially valuable plants. Despite this suborder great diversity, there are only few studies on the ultrastructure of these species spermatid. Thus, in this study, through transmission electron microscopy, we analyzed the development of spermiogenesis in the head and the flagellum of C. fuscus, L. gonagra, L. zonatus and S. fasciatus, with emphasis on the formation of the acrosome, of the centriole adjunct and on development of mitochondrial derivatives. The acrosome originates from the Golgi complex through the formation of a pro-acrosomal vesicle and concomitant to this process, is formed the centriole adjunct. The acrosome determines the anterior pole of the sperm and the centriole adjunct determines the posterior pole. It was also observed that at the beginning of the spermiogenic process, the mitochondria are scattered within the cytoplasm, then gradually fusing and originating the mitochondrial complex, which is subsequently divided into two substructures, called mitochondrial derivatives. They are positioned bilaterally to the axoneme, having the typical pattern arrangement of the microtubule, which is 9 + 9 + 2. The process of acrosomes and derivatives formation and the conformation of axoneme’s microtubules are typical of insects. Besides the pattern of axoneme’s microtubules, synapomorphic features of Heteroptera were found, as the presence of two paracrystalline structures in each mitochondrial derivative and the presence of bridges connecting derivatives mitochondrial to the axonemal microtubules, which are important tools to assist in phylogenetic and taxonomic analyzes.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Spermatogenesis of Zaprionus indianus and Zaprionus sepsoides (Diptera, Drosophilidae): Cytochemical, structural and ultrastructural characterization

Zaprionus indianus is a drosophilid native to the Afrotropical region that has colonized South America and exhibits a wide geographical distribution. In contrast, Z. sepsoides is restricted to certain African regions. The two species differ in the size of their testes, which are larger in Z. indianus than in Z. sepsoides. To better understand the biology and the degree of differentiation of these species, the current study evaluated spermatogenesis in males of different ages by conventional staining techniques and ultrastructural analysis. Spermatogenesis and the ultrastructure of spermatozoa were similar in the two species, and the diploid number was confirmed to be 2n = 12. A greater number of spermatozoa were observed in young Z. indianus (1-3 days old) compared to Z. sepsoides males, which showed a higher frequency of cells at the early stages of spermatogenesis. The head of the sperm was strongly marked by silver staining, lacto-acetic orcein and the Feulgen reaction; the P.A.S. reaction revealed glycogen granules in the testes of both species. Both species presented similar arrangement of microtubules (9+9+2), two mitochondrial derivatives of different size and 64 spermatozoa per bundle. Such similarity within the genus Zaprionus with other species of Drosophila, indicates that these structures are conserved in the family Drosophilidae. The differences observed the number and frequency of sperm cells in the early stages of spermatogenesis, between the young males of Z. indianus and Z. sepsoides, are features that may interfere with reproductive success and be related to the invasive potential of Z. indianus. © 2013, Sociedade Brasileira de Genética. Printed in Brazil.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Annexin 1: differential expression in tumor and mast cells in human larynx cancer.

Annexin 1 protein (ANXA1) expression was evaluated in tumor and mast cells in human larynx cancer and control epithelium. The effect of the exogenous ANXA1 (peptide Ac 2-26) was also examined during the cellular growth of the Hep-2 human larynx epidermoid carcinoma cell line. This peptide inhibited the proliferation of the Hep-2 cells within 144 hr. In surgical tissue specimens from 20 patients with larynx cancer, ultrastructural immunocytochemistry analysis showed in vivo down-regulation of ANXA1 expression in the tumor and increased in mast cells and Hep-2 cells treated with peptide Ac2-26. Combined in vivo and in vitro analysis demonstrated that ANXA1 plays a regulatory role in laryngeal cancer cell growth. We believe that a better understanding of the regulatory mechanisms of ANXA1 in tumor and mast cells may lead to future biological targets for the therapeutic intervention of human larynx cancer