Lysosomal Disorders Drive Susceptibility to Tuberculosis by Compromising Macrophage Migration.

A zebrafish genetic screen for determinants of susceptibility to Mycobacterium marinum identified a hypersusceptible mutant deficient in lysosomal cysteine cathepsins that manifests hallmarks of human lysosomal storage diseases. Under homeostatic conditions, mutant macrophages accumulate undigested lysosomal material, which disrupts endocytic recycling and impairs their migration to, and thus engulfment of, dying cells. This causes a buildup of unengulfed cell debris. During mycobacterial infection, macrophages with lysosomal storage cannot migrate toward infected macrophages undergoing apoptosis in the tuberculous granuloma. The unengulfed apoptotic macrophages undergo secondary necrosis, causing granuloma breakdown and increased mycobacterial growth. Macrophage lysosomal storage similarly impairs migration to newly infecting mycobacteria. This phenotype is recapitulated in human smokers, who are at increased risk for tuberculosis. A majority of their alveolar macrophages exhibit lysosomal accumulations of tobacco smoke particulates and do not migrate to Mycobacterium tuberculosis. The incapacitation of highly microbicidal first-responding macrophages may contribute to smokers' susceptibility to tuberculosis.

CLARITY and PACT-based imaging of adult zebrafish and mouse for whole-animal analysis of infections.

Visualization of infection and the associated host response has been challenging in adult vertebrates. Owing to their transparency, zebrafish larvae have been used to directly observe infection in vivo; however, such larvae have not yet developed a functional adaptive immune system. Cells involved in adaptive immunity mature later and have therefore been difficult to access optically in intact animals. Thus, the study of many aspects of vertebrate infection requires dissection of adult organs or ex vivo isolation of immune cells. Recently, CLARITY and PACT (passive clarity technique) methodologies have enabled clearing and direct visualization of dissected organs. Here, we show that these techniques can be applied to image host-pathogen interactions directly in whole animals. CLARITY and PACT-based clearing of whole adult zebrafish and Mycobacterium tuberculosis-infected mouse lungs enables imaging of mycobacterial granulomas deep within tissue to a depth of more than 1 mm. Using established transgenic lines, we were able to image normal and pathogenic structures and their surrounding host context at high resolution. We identified the three-dimensional organization of granuloma-associated angiogenesis, an important feature of mycobacterial infection, and characterized the induction of the cytokine tumor necrosis factor (TNF) within the granuloma using an established fluorescent reporter line. We observed heterogeneity in TNF induction within granuloma macrophages, consistent with an evolving view of the tuberculous granuloma as a non-uniform, heterogeneous structure. Broad application of this technique will enable new understanding of host-pathogen interactions in situ.

Transcriptional profiling of chronic clinical hepatic schistosomiasis japonica indicates reduced metabolism and immune responses

Schistosomiasis is a significant cause of human morbidity and mortality. We performed a genome-wide transcriptional survey of liver biopsies obtained from Chinese patients with chronic schistosomiasis only, or chronic schistosomiasis with a current or past history of viral hepatitis B. Both disease groups were compared with patients with no prior history or indicators of any liver disease. Analysis showed in the main, downregulation in gene expression, particularly those involved in signal transduction via EIF2 signalling and mTOR signalling, as were genes associated with cellular remodelling. Focusing on immune associated pathways, genes were generally downregulated. However, a set of three genes associated with granulocytes, MMP7, CLDN7, CXCL6 were upregulated. Differential gene profiles unique to schistosomiasis included the gene Granulin which was decreased despite being generally considered a marker for liver disease, and IGBP2 which is associated with increased liver size, and was the most upregulated gene in schistosomiasis only patients, all of which presented with hepatomegaly. The unique features of gene expression, in conjunction with previous reports in the murine model of the cellular composition of granulomas, granuloma formation and recovery, provide an increased understanding of the molecular immunopathology and general physiological processes underlying hepatic schistosomiasis.

Suppression of the Insulin Receptors in Adult Schistosoma japonicum Impacts on Parasite Growth and Development: Further Evidence of Vaccine Potential

To further investigate the importance of insulin signaling in the growth, development, sexual maturation and egg production of adult schistosomes, we have focused attention on the insulin receptors (SjIRs) of Schistosoma japonicum, which we have previously cloned and partially characterised. We now show, by Biolayer Interferometry, that human insulin can bind the L1 subdomain (insulin binding domain) of recombinant (r)SjIR1 and rSjIR2 (designated SjLD1 and SjLD2) produced using the Drosophila S2 protein expression system. We have then used RNA interference (RNAi) to knock down the expression of the SjIRs in adult S. japonicum in vitro and show that, in addition to their reduced transcription, the transcript levels of other important downstream genes within the insulin pathway, associated with glucose metabolism and schistosome fecundity, were also impacted substantially. Further, a significant decrease in glucose uptake was observed in the SjIR-knockdown worms compared with luciferase controls. In vaccine/challenge experiments, we found that rSjLD1 and rSjLD2 depressed female growth, intestinal granuloma density and faecal egg production in S. japonicum in mice presented with a low dose challenge infection. These data re-emphasize the potential of the SjIRs as veterinary transmission blocking vaccine candidates against zoonotic schistosomiasis japonica in China and the Philippines.

Vänskap är magisk : En reparativ läsning av samtida svensk seriekonst

In this thesis, I do a reparative reading of contemporary Swedish comics, that uses feminine signifiers, both in their imagery and writing. A crucial point of departure, in this thesis, is how literature can ”do” theory and be seen as a way of creating knowledge. I explore what the comics does to me, as a reader, and how. I experiment with the form and style of academic writing in order to clearly define my position and situate the production of knowledge. By focusing on the affects and nourishment, that the comics contain, I try to imagine a feminist other, with the help of my figuration: Nietzsche Minaj, and my imaginary utopian place: ”mitt flick(tionella) rum”. I conclude, that the comics both reproduce and transform feminine signifiers, while challenging the idea of dichotomies, in the spirit of gurlesque theory.

Prevalência de alterações fundoscópicas em estudantes na cidade de Natal/Brasil

Objetivo: Determinar a prevalência de alterações fundoscópicas em estudantes de escolas das redes pública e privada de Natal-RN. Métodos: Avaliação oftalmológica foi realizada em 990 alunos, de 5 a 21 anos, matriculados nas escolas das redes públicas e privada do município de Natal- RN, que estiveram cursando alguma série do ensino fundamental ou médio, no período de 03 a 06 de 2001. Resultados: Alterações fundoscópicas foram observadas em 5,3% dos estudantes. As anormalidades encontradas, por ordem de freqüência, foram: branco sem pressão, 1,0%; cicatriz de retinocoroidite sugestiva de toxoplasmose, 1,0%; atrofia do epitélio pigmentado da retina, 0,8%; nevos da coróide, 0,4%; escavação da cabeça do nervo óptico aumentada, 0,4%; degeneração em treliça, 0,3%; buraco operculado, 0,2%; fundus miópico, 0,2%; tortuosidade vascular aumentada, 0,2%; granuloma sugestivo de toxocaríase, 0,2%; hipoplasia da cabeça do nervo óptico, 0,1%; persistência da artéria hialoidea, 0,1%; persistência de fibras de mielina, 0,1%; retina sal e pimenta, 0,1%; retinosquise, 0,1%. Conclusão: Houve uma baixa prevalência de alterações fundoscópicas na população estudada

Prevalência de alterações fundoscópicas em estudantes na cidade de Natal-Brasil

Objetivo: Determinar a prevalência de alterações fundoscópicas em estudantes de escolas das redes pública e privada de Natal-RN. Métodos: Avaliação oftalmológica foi realizada em 990 alunos, de 5 a 21 anos, matriculados nas escolas das redes públicas e privada do município de Natal-RN, que estiveram cursando alguma série do ensino fundamental ou médio, no período de 03 a 06 de 2001. Resultados: Alterações fundoscópicas foram observadas em 5,3% dos estudantes. As anormalidades encontradas, por ordem de freqüência, foram: branco sem pressão, 1,0%; cicatriz de retinocoroidite sugestiva de toxoplasmose, 1,0%; atrofia do epitélio pigmentado da retina, 0,8%; nevos da coróide, 0,4%; escavação da cabeça do nervo óptico aumentada, 0,4%; degeneração em treliça, 0,3%; buraco operculado, 0,2%; fundus miópico, 0,2%; tortuosidade vascular aumentada, 0,2%; granuloma sugestivo de toxocaríase, 0,2%; hipoplasia da cabeça do nervo óptico, 0,1%; persistência da artéria hialoidea, 0,1%; persistência de fibras de mielina, 0,1%; retina sal e pimenta, 0,1%; retinosquise, 0,1%. Conclusão: Houve uma baixa prevalência de alterações fundoscópicas na população estudada

Masson’s tumor: a soft tissue tumor simulating a tendon cyst. Case report

Introduction. Intravascular papillary endothelial hyperplasia (Masson's hemangioma or Masson’s tumor) is a benign vascular disease with an exuberant endothelial proliferation in normal blood vessels. Although relatively uncommon, its correct diagnosis is important because it can clinically be like both benign lesions and malignant neoplasms. We present a case of intravascular proliferative endothelial hyperplasia simulating a tendon cyst both clinically and on ultrasound. Case report. A 74-year old Caucasian female presented with a 4-month history of soreness and swelling in the fourth finger of the right hand. Ultrasound showed an oval mass with fluid content, referred to a tendon cyst. A wide surgical excision was subsequently performed. The final histological diagnosis was Masson’s tumor. Discussion. The pathogenesis of intravascular papillary endothelial hyperplasia is still unclear but the exuberant endothelial cell proliferation might be stimulated by an autocrine loop of endothelial basic fibroblast growth factor (bFGF) secretion. There are three types of papillary endothelial hyperplasia: primary, or intravascular; secondary, or mixed; and extravascular. The main differential diagnosis is against pyogenic granuloma, Kaposi sarcoma, hemangioma, and angiosarcoma. Conclusions. Masson's tumor can be like both benign lesions and malignant neoplasms clinically and on ultrasound. For this reason, the right diagnosis can be made only by histology, which reveals a papillary growth composed of hyperplastic endothelial cells supported by delicate fibrous stalks entirely confined within the vascular lumen.

Uma abordagem sobre infecções endodônticas

As infeções endodônticas envolvem a invasão e multiplicação de microrganismos na polpa dentária e tecidos periapicais sendo responsáveis por dois tipos de patologias: as patologias pulpares e as patologias periapicais. Relativamente às patologias pulpares destacam-se a pulpite reversível, a pulpite irreversível e a necrose pulpar. Quanto às patologias periapicais, destacam-se o abcesso apical agudo, o abcesso apical crónico, a periodontite apical aguda, a periodontite apical crónica, o granuloma perirradicular e o quisto perirradicular. As doenças pulpares e periapicais apresentam manifestações clínicas diferentes que, em conjunto com os sinais e sintomas manifestados pelo paciente permitem diagnosticar o tipo de infeção endodôntica. As infeções endodônticas estão associadas a uma elevada diversidade de bactérias, sendo frequentemente intituladas de infeções endodônticas polimicrobianas. Sabe-se que os microrganismos são a causa principal das doenças pulpares e periapicais e, por esse motivo, o objetivo principal do Tratamento Endodôntico consiste na eliminação dos microrganismos e prevenção da re-infeção. O tratamento das infeções endodônticas baseia-se na preparação químico-mecânica do sistema de canais radiculares – instrumentação e irrigação – seguida da obturação e culminando com a restauração definitiva ou tratamento reabilitador. Este trabalho tem como objetivos adquirir um conhecimento mais amplo relativamente aos tipos de infeções endodônticas, à realização dos diversos diagnósticos e, principalmente, às várias opções de tratamento, disponíveis na área da Endodontia. Para tal foi realizada uma pesquisa bibliográfica baseada em artigos científicos, publicados nas bases de dados PubMed, Scielo e Science Direct bem como em alguns livros relacionados com o tema.

Bartonelosis sistémica con lesiones hepatoesplénicas en un paciente inmunocompetente: Presentación de un caso y revisión de la bibliografía

Resumen La enfermedad por arañazo de gato (EAG) es una enfermedad infecciosa causada por Bartonella henselae. Esta enfermedad es de evolución habitualmente benigna, el beneficio del tratamiento antibiótico no está claramente demostrado, sin embargo algunos pacientes desarrollan formas atípicas y más graves con compromiso sistémico como la formación de granulomas hepáticos y esplénicos. Se presenta el caso clínico de una niña de 10 años con granulomas hepáticos que fue asistida en el Centro Hospitalario Pereira Rossell por fiebre prolongada, astenia, dolor abdominal y adelgazamiento. Lo que obliga a investigar la etiología y descartar diagnósticos diferenciales y proponer alternativas terapéuticas. En ese momento no disponíamos de estudios serológicos para la búsqueda etiológica en el país lo cual generó dificultades diagnósticas en esta paciente. Finalmente se pudo obtener la confirmación serológica de EAG de la paciente al enviar la muestra al Hospital de Niños Ricardo Gutiérrez en Buenos Aires prácticamente un mes después de iniciado los síntomas. Fue tratada inicialmente con azitromicina vía oral 10 mg/kg/día por 10 días y dada la tórpida evolución se utiliza un segundo plan antibiótico que incluyó claritromicina 15 mg/kg/día más rifampicina 20 mg/kg/día por 6 semanas. La paciente evolucionó favorablemente, presentando disminución progresiva del tamaño de los granulomas hepáticos y esplénicos. Discusión: Los granulomas hepatoesplénicos constituyen una forma de presentación poco frecuente pero orientadora de la EAG. La evidencia en relación a su tratamiento es escasa y deriva de series de casos. Es posible que los pacientes con manifestaciones sistémicas requieran tratamientos combinados y prolongados.

Prevalência de alterações fundoscópicas em estudantes na cidade de Natal/Brasil

Objetivo: Determinar a prevalência de alterações fundoscópicas em estudantes de escolas das redes pública e privada de Natal-RN. Métodos: Avaliação oftalmológica foi realizada em 990 alunos, de 5 a 21 anos, matriculados nas escolas das redes públicas e privada do município de Natal- RN, que estiveram cursando alguma série do ensino fundamental ou médio, no período de 03 a 06 de 2001. Resultados: Alterações fundoscópicas foram observadas em 5,3% dos estudantes. As anormalidades encontradas, por ordem de freqüência, foram: branco sem pressão, 1,0%; cicatriz de retinocoroidite sugestiva de toxoplasmose, 1,0%; atrofia do epitélio pigmentado da retina, 0,8%; nevos da coróide, 0,4%; escavação da cabeça do nervo óptico aumentada, 0,4%; degeneração em treliça, 0,3%; buraco operculado, 0,2%; fundus miópico, 0,2%; tortuosidade vascular aumentada, 0,2%; granuloma sugestivo de toxocaríase, 0,2%; hipoplasia da cabeça do nervo óptico, 0,1%; persistência da artéria hialoidea, 0,1%; persistência de fibras de mielina, 0,1%; retina sal e pimenta, 0,1%; retinosquise, 0,1%. Conclusão: Houve uma baixa prevalência de alterações fundoscópicas na população estudada

Prevalência de alterações fundoscópicas em estudantes na cidade de Natal-Brasil

Objetivo: Determinar a prevalência de alterações fundoscópicas em estudantes de escolas das redes pública e privada de Natal-RN. Métodos: Avaliação oftalmológica foi realizada em 990 alunos, de 5 a 21 anos, matriculados nas escolas das redes públicas e privada do município de Natal-RN, que estiveram cursando alguma série do ensino fundamental ou médio, no período de 03 a 06 de 2001. Resultados: Alterações fundoscópicas foram observadas em 5,3% dos estudantes. As anormalidades encontradas, por ordem de freqüência, foram: branco sem pressão, 1,0%; cicatriz de retinocoroidite sugestiva de toxoplasmose, 1,0%; atrofia do epitélio pigmentado da retina, 0,8%; nevos da coróide, 0,4%; escavação da cabeça do nervo óptico aumentada, 0,4%; degeneração em treliça, 0,3%; buraco operculado, 0,2%; fundus miópico, 0,2%; tortuosidade vascular aumentada, 0,2%; granuloma sugestivo de toxocaríase, 0,2%; hipoplasia da cabeça do nervo óptico, 0,1%; persistência da artéria hialoidea, 0,1%; persistência de fibras de mielina, 0,1%; retina sal e pimenta, 0,1%; retinosquise, 0,1%. Conclusão: Houve uma baixa prevalência de alterações fundoscópicas na população estudada

Gestation and breastfeeding in schistosomotic mothers differently modulate the immune response of adult offspring to postnatal Schistosoma mansoni infection

Schistosoma mansoni antigens in the early life alter homologous and heterologous immunity during postnatal infections. We evaluate the immunity to parasite antigens and ovalbumin (OA) in adult mice born/suckled by schistosomotic mothers. Newborns were divided into: born (BIM), suckled (SIM) or born/suckled (BSIM) in schistosomotic mothers, and animals from noninfected mothers (control). When adults, the mice were infected and compared the hepatic granuloma size and cellularity. Some animals were OA + adjuvant immunised. We evaluated hypersensitivity reactions (HR), antibodies levels (IgG1/IgG2a) anti-soluble egg antigen and anti-soluble worm antigen preparation, and anti-OA, cytokine production, and CD4+FoxP3+T-cells by splenocytes. Compared to control group, BIM mice showed a greater quantity of granulomas and collagen deposition, whereas SIM and BSIM presented smaller granulomas. BSIM group exhibited the lowest levels of anti-parasite antibodies. For anti-OA immunity, immediate HR was suppressed in all groups, with greater intensity in SIM mice accompanied of the remarkable level of basal CD4+FoxP3+T-cells. BIM and SIM groups produced less interleukin (IL)-4 and interferon (IFN)-g. In BSIM, there was higher production of IL-10 and IFN-γ, but lower levels of IL-4 and CD4+FoxP3+T-cells. Thus, pregnancy in schistosomotic mothers intensified hepatic fibrosis, whereas breastfeeding diminished granulomas in descendants. Separately, pregnancy and breastfeeding could suppress heterologous immunity; however, when combined, the responses could be partially restored in infected descendants.

Cytomegalovirus and Langerhans Cell Histiocytosis: Is There a Link?

Background: Langerhans cell histiocytosis is a rare proliferative histiocytic disease of unknown etiology. Histologically, it is characterized by granuloma-like proliferation of Langerhans-type dendritic cells derived from bone marrow. Many investigators have suggested the possible role of viruses such as Epstein-Barr virus, human herpesvirus-6 (HHV-6), herpes simplex virus (HSV) types 1 and 2, and Cytomegalovirus in the pathogenesis of Langerhans cell histiocytosis. Objectives: In this study, we have investigated the presence of Cytomegalovirus in Langerhans cell histiocytosis in Iranian children. Patients and Methods: In this retrospective study, we have investigated the presence of Cytomegalovirus DNA expression, using paraffin-embedded tissue samples of 30 patients with Langerhans cell histiocytosis and 30 age and site-matched controls by qualitative Polymerase Chain Reaction (PCR) method. Results: No significant difference in prevalence of Cytomegalovirus presence between patients and controls was found. Cytomegalovirus was found by qualitative PCR in only 2 (6.66%) out of 30 patients and in 1 (3.3%) of 30 control samples with a P value of 1 (1.00 > 0.05) using chi-square test with OR: 2.07; 95% CI of OR: 0.18 - 24.15. Conclusions: Our findings do not support the hypothesis of a possible role for Cytomegalovirus in the pathogenesis of Langerhans cell histiocytosis.

Discovery of stable and variable differences in the Mycobacterium avium subsp. paratuberculosis type I, II, and III genomes by pan-genome microarray analysis

Mycobacterium avium subsp. paratuberculosis is an important animal pathogen widely disseminated in the environment that has also been associated with Crohn's disease in humans. Three M. avium subsp. paratuberculosis genomotypes are recognized, but genomic differences have not been fully described. To further investigate these potential differences, a 60-mer oligonucleotide microarray (designated the MAPAC array), based on the combined genomes of M. avium subsp. paratuberculosis (strain K-10) and Mycobacterium avium subsp. hominissuis (strain 104), was designed and validated. By use of a test panel of defined M. avium subsp. paratuberculosis strains, the MAPAC array was able to identify a set of large sequence polymorphisms (LSPs) diagnostic for each of the three major M. avium subsp. paratuberculosis types. M. avium subsp. paratuberculosis type II strains contained a smaller genomic complement than M. avium subsp. paratuberculosis type I and M. avium subsp. paratuberculosis type III genomotypes, which included a set of genomic regions also found in M. avium subsp. hominissuis 104. Specific PCRs for genes within LSPs that differentiated M. avium subsp. paratuberculosis types were devised and shown to accurately screen a panel (n = 78) of M. avium subsp. paratuberculosis strains. Analysis of insertion/deletion region INDEL12 showed deletion events causing a reduction in the complement of mycobacterial cell entry genes in M. avium subsp. paratuberculosis type II strains and significantly altering the coding of a major immunologic protein (MPT64) associated with persistence and granuloma formation. Analysis of MAPAC data also identified signal variations in several genomic regions, termed variable genomic islands (vGIs), suggestive of transient duplication/deletion events. vGIs contained significantly low GC% and were immediately flanked by insertion sequences, integrases, or short inverted repeat sequences. Quantitative PCR demonstrated that variation in vGI signals could be associated with colony growth rate and morphology.