The pattern of immune cell infiltration in chromoblastomycosis: involvement of macrophage inflammatory protein-1 alpha/CCL3 and fungi persistence

Chromoblastomycosis (CR) is a subcutaneous chronic mycosis characterized by a granulomatous inflammatory response. However, little is known regarding the pattern of leukocyte subsets in CR and the pathways involved in their recruitment. The objective of this study was to assess the cellular subsets, chemokine, chemokine receptors and enzymes in CR. The inflammatory infiltrate was characterized by immunohistochemistry using antibodies against macrophages (CD68), Langerhans'cells (S100), lymphocytes (CD3, CD4, CD8, CD45RO, CD20 and CD56) and neutrophils (CD15). The expression of MIP-1alpha (Macrophage inflammatory protein-1alpha), chemokine receptors (CXCR3 and CCR1) and enzymes (superoxide dismutase-SOD and nitric oxide synthase-iNOS) was also evaluated by the same method. We observed an increase in all populations evaluated when compared with the controls. Numbers of CD15+ and CD56+ were significantly lower than CD3+, CD4+, CD20+ and CD68+ cells. Statistical analysis revealed an association of fungi numbers with CD3, CD45RO and iNOS-positive cells. Furthermore, MIP-1alpha expression was associated with CD45RO, CD68, iNOS and CXCR3. Our results suggest a possible role of MIP-1alpha and fungi persistence in the cell infiltration in CR sites.

Characterization of a clone from an adult worm cDNA library selected with anti-Schistosoma mansoni human antibodies dissociated from immune complexes: a preliminary report

Considering the scarcity of defined antigens, actually useful and reliable for use in the field studies, we propose an alternative method for selection of cDNA clones with potential use in the diagnosis of schistosomiasis. Human antibodies specific to a protein fraction of 31/32 kDa (Sm31/32), dissociated from immune complexes, are used for screening of clones from an adult worm cDNA library. Partial sequencing of five clones, selected through this strategy, showed to be related to Schistosoma mansoni: two were identified as homologous to heat shock protein 70, one to glutathione S-transferase, one to homeodomain protein, and one to a previously described EST (expressed sequence tag) of S. mansoni. This last clone was the most consistently reactive during the screening process with the anti-Sm31/32 antibodies dissociated from the immune complexes. The complete sequence of this clone was obtained and the translation data yielded only one ORF (open reading frame) that code for a protein with 57 amino acids. Based on this amino acid sequence two peptides were chemically synthesized and evaluated separately against a pool of serum samples from schistosomiasis patients and non-schistosomiasis individuals. Both peptides showed strong reactivity only against the positive pool, suggesting that these peptides may be useful as antigens for the diagnosis of schistosomiasis mansoni.

Molecular detection of "Candidatus Mycoplasma haemominutum" in a lion (Panthera leo) from a brazilian zoological garden

Although Mycoplasma haemofelis and "Candidatus Mycoplasma haemominutum" infections have been reported in wild cats from United States, their presence among native and captive wild cats in Brazil is still unknown. A 12 year old healthy male lion (Panthera leo) from the Zoological Garden of Curitiba, Brazil was anesthetized for transportation and dental evaluation. A blood sample was obtained for a complete blood cell count (CBC) and PCR analysis. DNA was extracted and fragments of Mycoplasma haemofelis and "Candidatus Mycoplasma haemominutum" 16S ribosomal RNA gene were amplified in PCR assays. CBC results were within reference intervals. A weak band of 192 pb for "Candidatus Mycoplasma haemominutum" was observed, and no band was amplified from Mycoplasma haemofelis reaction. A weak PCR band associated with normal CBC results and without visible parasitemia or clinical signs may suggest a chronic subclinical infection with "Candidatus Mycoplasma haemominutum". The lack of clinical signs may also represent the low pathogenicity of this organism; however, it is noteworthy that immune suppression caused by management and/or corticoids treatment may induce parasitemia and anemia in this animal. This detection suggests further studies in captive wild cats in Brazilian Zoological Gardens.

Enzymatic and hemolytic activities of Candida dubliniensis strains

Candida dubliniensis is an opportunistic yeast that has been recovered from several body sites in many populations; it is most often recovered from the oral cavities of human immunodeficiency virus-infected patients. Although extensive studies on epidemiology and phylogeny of C. dubliniensis have been performed, little is known about virulence factors such as exoenzymatic and hemolytic activities. In this study we compared proteinase, hyaluronidase, chondroitin sulphatase and hemolytic activities in 18 C. dubliniensis and 30 C. albicans strains isolated from AIDS patients. C. albicans isolates produced higher amounts of proteinase than C. dubliniensis (p < 0.05). All the tested C. dubliniensis strains expressed hyaluronidase and chondroitin sulphatase activities, but none of them were significantly different from those observed with C. albicans (p > 0.05). Hemolytic activity was affected by CaCl2; when this component was absent, we did not notice any significant difference between C. albicans and C. dubliniensis hemolytic activities. On the contrary, when we added 2.5 g% CaCl2, the hemolytic activity was reduced on C. dubliniensis and stimulated on C. albicans tested strains (p < 0.05).

Soft tissue abscess and lymphadenitis due to Mycobacterium avium Complex as an expression of immune reconstitution inflammatory syndrome after a second scheme of highly active antiretroviral therapy

Immune reconstitution inflammatory syndrome (IRIS) is an atypical and unexpected reaction related to highly active antiretroviral therapy (HAART) in human immunodeficiency virus (HIV) infected patients. IRIS includes an atypical response to an opportunistic pathogen (generally Mycobacterium tuberculosis, Mycobacterium avium complex, cytomegalovirus and herpes varicella-zoster), in patients responding to HAART with a reduction of plasma viral load and evidence of immune restoration based on increase of CD4+ T-cell count. We reported a case of a patient with AIDS which, after a first failure of HAART, developed a subcutaneous abscess and supraclavicular lymphadenitis as an expression of IRIS due to Mycobacterium avium complex after starting a second scheme of HAART.

A comparison of the immune parameters of dogs infected with visceral leishmaniasis using Western blot and neutralization techniques

The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage) it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.

Parasitoses Intestinais Como Causa de Anemia na Gravidez: Três Casos Clínicos Ilustrativos

Os autores apresentam três casos clínicos de parasitoses, diagnosticadas no contexto de anemia. A propósito destas situações é feita uma revisão teórica das infestações em causa - shistosomíase, ancilostomíase, giardíase, trichiuríase e ascaridíase.

Pode a Presença de Anemia na Admissão Melhorar a Capacidade Preditiva do Score GRACE para Mortalidade a Curto e Médio Prazo após Síndrome Coronária Aguda?

Introdução: Nos doentes admitidos com síndrome coronária aguda (SCA), a presença de anemia é um fator predizente de prognóstico. Contudo, os diversos scores de risco após SCA não incluem este fator. Objetivos: Avaliar se a presença de anemia na admissão em doentes com SCA tem valor acrescido relativamente ao score GRACE na predição de mortalidade a curto e médio prazo. Métodos: Entre janeiro 2005 e dezembro 2008, avaliaram-se os doentes admitidos consecutivamente na nossa Unidade de Cuidados Intensivos por SCA e incluídos no registo de SCA do centro. Em todos os doentes foram colhidos dados demográficos, antropométricos, fatores de risco para doença coronária, dados clínicos e laboratoriais da admissão, incluindo hemoglobina. Foram identificados os doentes com anemia (hemoglobina < 12 g/dL nas mulheres e < 13 g/dL nos homens). Os doentes foram divididos em risco baixo, intermédio e alto: < 126, 126-154 e > 154 para o score GRACE, respetivamente. Analisou-se a ocorrência de morte intra-hospitalar, aos 30 dias e ao primeiro ano de seguimento. Resultados: Incluíram-se 1423 doentes, com idade média de 64 ± 13 anos, 69% do sexo masculino,identificando-se a presenc¸a de anemia na admissão em 27,7% dos doentes. Estes doentes eram mais idosos, com predomínio do sexo feminino, mais hipertensos e diabéticos, maior número com história prévia de enfarte, com pior classe de Killip na admissão e score GRACE mais alto. Pelo contrário, eram menos fumadores, com menor apresentação como enfarte com supradesnivelamento ST e receberam menos bloqueadores beta, estatinas e angioplastia coronária. Tiveram também mais complicações hemorrágicas durante o internamento. A mortalidade intra-hospitalar (10 versus 4%), aos 30 dias (12 versus 5%) e ao primeiro ano (15 versus 6%) foram superiores no grupo com anemia (p < 0,001). Na análise bivariada, a presença de anemia é fator predizente de mortalidade intra-hospitalar (OR 2,46, IC 95% 1,57-3,85, p < 0,001), aos 30 dias (OR 2,47, IC 95% 1,65-3,69, p < 0,001) e ao primeiro ano (OR 2,66, IC 95% 1,83-3,86, p < 0,001), não se mantendo, contudo, esta associação após ajuste para outras variáveis. Associando a presença de anemia ao score GRACE, diferencia apenas para a mortalidade ao primeiro ano(com maior mortalidade) os grupos de risco intermédio e alto do score GRACE (6,7 versus 2,3%, p = 0,024; 23,4 versus 15,6%, p = 0,022, respetivamente), com uma tendência para diferenciar a mortalidade aos 30 dias no grupo de risco alto de score (19,6 versus 13,5%, p = 0,056). Conclusão: Os nossos dados confirmam que a anemia é um fator predizente importante de mortalidade a curto e médio prazo após SCA, contudo, não significativo quando ajustado ou incluído no score GRACE. Contudo, a sua combinação com o score GRACE pode melhorar a estratificação de risco, em particular no alto risco.

Leprosy and Kaposi Sarcoma Presenting as an Immune Reconstitution Inflammatory Syndrome in a Patient with AIDS

The simultaneous presence of infectious organisms within cutaneous lesions of Kaposi sarcoma in persons with AIDS has been demonstrated. We describe a patient with concurrent leprosy and Kaposi sarcoma presenting as an immune reconstitution inflammatory syndrome in the setting of AIDS.

Cytokine profile in Montenegro skin test of patients with localized cutaneous and mucocutaneous leishmaniasis

American tegumentary leishmaniasis presents as two major clinical forms: localized cutaneous leishmaniasis (LCL) and mucocutaneous leishmaniasis (MCL). The immune response in leishmaniasis is efficiently evaluated by the response to Leishmania antigen through the Montenegro skin test (MST). Both LCL and MCL present positive response to MST, indicating that the patients present cell-mediated immunity against the parasite - Leishmania. In spite of the presence of immunity in MCL, this is not sufficient to stop disease progression and prevent resistance to treatment. In this study we demonstrated interleukin (IL) 2, 4, 5 and interferon (IFN) gamma expression in biopsies of MST of ten patients with American tegumentary leishmaniasis. The obtained results were compared between LCL (n = 5) and MCL (n = 5) patients. The MST of MCL patients displayed a higher expression of IL-2, IL-4 and IL-5, in comparison to LCL. There was no significant difference in IFN-gamma expression between groups. The obtained results suggest the role of IL-4 and IL-5 in the maintenance of the immunopathogenic mechanism of the destructive lesions that characterize MCL.

Protective effect of C. sativa leaf extract against UV mediated-DNA damage in a human keratinocyte cell line

Toxic effects of ultraviolet (UV) radiation on skin include protein and lipid oxidation, and DNA damage. The latter is known to play a major role in photocarcinogenesis and photoaging. Many plant extracts and natural compounds are emerging as photoprotective agents. Castanea sativa leaf extract is able to scavenge several reactive species that have been associated to UV-induced oxidative stress. The aim of this work was to analyze the protective effect of C. sativa extract (ECS) at different concentrations (0.001, 0.01, 0.05 and 0.1 μg/mL) against the UV mediated-DNA damage in a human keratinocyte cell line (HaCaT). For this purpose, the cytokinesis-block micronucleus assay was used. Elucidation of the protective mechanism was undertaken regarding UV absorption, influence on 1O2 mediated effects or NRF2 activation. ECS presented a concentration-dependent protective effect against UV-mediated DNA damage in HaCaT cells. The maximum protection afforded (66.4%) was achieved with the concentration of 0.1 μg/mL. This effect was found to be related to a direct antioxidant effect (involving 1O2) rather than activation of the endogenous antioxidant response coordinated by NRF2. Electrochemical studies showed that the good antioxidant capacity of the ECS can be ascribed to the presence of a pool of different phenolic antioxidants. No genotoxic or phototoxic effects were observed after incubation of HaCaT cells with ECS (up to 0.1 μg/mL). Taken together these results reinforce the putative application of this plant extract in the prevention/minimization of UV deleterious effects on skin.

Anemia Hemolítica Auto-Imune - Caso Clínico

A anemia hemolitica é uma entidade caracterizada por destruição dos glóbulos vermelhos mediada por auto-anticorpos. Os mecanismos fisiopatológicos ainda nao estão completamente compreendidos e a terapêutica é controversa.Os autores apresentam este caso pela sua particular evolução/resposta terapêutica.

Enhancing Agent Mediated Electronic Markets with Ontology Matching Services and Social Network Support

Os Mercados Eletrónicos atingiram uma complexidade e nível de sofisticação tão elevados, que tornaram inadequados os modelos de software convencionais. Estes mercados são caracterizados por serem abertos, dinâmicos e competitivos, e constituídos por várias entidades independentes e heterogéneas. Tais entidades desempenham os seus papéis de forma autónoma, seguindo os seus objetivos, reagindo às ocorrências do ambiente em que se inserem e interagindo umas com as outras. Esta realidade levou a que existisse por parte da comunidade científica um especial interesse no estudo da negociação automática executada por agentes de software [Zhang et al., 2011]. No entanto, a diversidade dos atores envolvidos pode levar à existência de diferentes conceptualizações das suas necessidades e capacidades dando origem a incompatibilidades semânticas, que podem prejudicar a negociação e impedir a ocorrência de transações que satisfaçam as partes envolvidas. Os novos mercados devem, assim, possuir mecanismos que lhes permitam exibir novas capacidades, nomeadamente a capacidade de auxiliar na comunicação entre os diferentes agentes. Pelo que, é defendido neste trabalho que os mercados devem oferecer serviços de ontologias que permitam facilitar a interoperabilidade entre os agentes. No entanto, os humanos tendem a ser relutantes em aceitar a conceptualização de outros, a não ser que sejam convencidos de que poderão conseguir um bom negócio. Neste contexto, a aplicação e exploração de relações capturadas em redes sociais pode resultar no estabelecimento de relações de confiança entre vendedores e consumidores, e ao mesmo tempo, conduzir a um aumento da eficiência da negociação e consequentemente na satisfação das partes envolvidas. O sistema AEMOS é uma plataforma de comércio eletrónico baseada em agentes que inclui serviços de ontologias, mais especificamente, serviços de alinhamento de ontologias, incluindo a recomendação de possíveis alinhamentos entre as ontologias dos parceiros de negociação. Este sistema inclui também uma componente baseada numa rede social, que é construída aplicando técnicas de análise de redes socias sobre informação recolhida pelo mercado, e que permite melhorar a recomendação de alinhamentos e auxiliar os agentes na sua escolha. Neste trabalho são apresentados o desenvolvimento e implementação do sistema AEMOS, mais concretamente: • É proposto um novo modelo para comércio eletrónico baseado em agentes que disponibiliza serviços de ontologias; • Adicionalmente propõem-se o uso de redes sociais emergentes para captar e explorar informação sobre relações entre os diferentes parceiros de negócio; • É definida e implementada uma componente de serviços de ontologias que é capaz de: • o Sugerir alinhamentos entre ontologias para pares de agentes; • o Traduzir mensagens escritas de acordo com uma ontologia em mensagens escritas de acordo com outra, utilizando alinhamentos previamente aprovados; • o Melhorar os seus próprios serviços recorrendo às funcionalidades disponibilizadas pela componente de redes sociais; • É definida e implementada uma componente de redes sociais que: • o É capaz de construir e gerir um grafo de relações de proximidade entre agentes, e de relações de adequação de alinhamentos a agentes, tendo em conta os perfis, comportamento e interação dos agentes, bem como a cobertura e utilização dos alinhamentos; • o Explora e adapta técnicas e algoritmos de análise de redes sociais às várias fases dos processos do mercado eletrónico. A implementação e experimentação do modelo proposto demonstra como a colaboração entre os diferentes agentes pode ser vantajosa na melhoria do desempenho do sistema e como a inclusão e combinação de serviços de ontologias e redes sociais se reflete na eficiência da negociação de transações e na dinâmica do mercado como um todo.