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BACKGROUND Dendritic cells (DCs) specific intercellular adhesion molecule (ICAM)-3-grabbing non integrin receptor (DC-SIGN) binds to subgenera Leishmania promastigotes mediating its interaction with DC and neutrophils, potentially influencing the infection outcome. OBJECTIVES In this work, we investigated whether DC-SIGN receptor is expressed in cells from cutaneous leishmaniasis (CL) lesions as well as the in vitro binding pattern of Leishmania (Viannia) braziliensis (Lb) and L. (L.) amazonensis (La) promastigotes. METHODS DC-SIGN receptor was labeled by immunohistochemistry in cryopreserved CL tissue fragments. In vitro binding assay with CFSE-labeled Lb or La promastigotes and RAJI-transfecting cells expressing DC-SIGN (DC-SIGNPOS) or mock-transfected (DC-SIGNNEG) were monitored by flow cytometry at 2 h, 24 h and 48 h in co-culture. RESULTS In CL lesion infiltrate, DC-SIGNPOS cells were present in the dermis and near the epidermis. Both Lb and La bind to DC-SIGNPOS cells, while binding to DC-SIGNNEG was low. La showed precocious and higher affinity to DC-SIGNhi population than to DC-SIGNlow, while Lb binding was similar in these populations. CONCLUSION Our results demonstrate that DC-SIGN receptor is present in L. braziliensis CL lesions and interact with Lb promastigotes. Moreover, the differences in the binding pattern to Lb and La suggest DC-SIGN can influence in a difference way the intake of the parasites at the first hours after Leishmania infection. These results raise the hypothesis that DC-SIGN receptor could participate in the immunopathogenesis of American tegumentary leishmaniasis accounting for the differences in the outcome of the Leishmania spp. infection.

Background. Cytokines and growth factors involved in the tissue inflammatory process influence the outcome of Leishmania infection. Insulin-like growth factor (IGF-I) constitutively present in the skin may participate in the inflammatory process and parasite-host interaction. Previous work has shown that preincubation of Leishmania (Leishmania) amazonensis with recombinant IGF-I induces accelerated lesion development. However, in human cutaneous leishmaniasis (CL) pathogenesis, it is more relevant to the persistent inflammatory process than progressive parasite proliferation. In this context, we aimed to investigate whether IGF-I was present in the CL lesions and if this factor may influence the lesions’ development acting on parasite growth and/or on the inflammatory/healing process. Methodology. Fifty-one CL patients’ skin lesion samples from endemic area of L. (Viannia) braziliensis infection were submitted to histopathological analysis and searched for Leishmania and IGF-I expression by immunohistochemistry. Results. In human CL lesions, IGF-I was observed preferentially in the late lesion (more than 90 days), and the percentage of positive area for IGF-I was positively correlated with duration of illness ( r = 0.42 , P < 0.05 ). IGF-I was highly expressed in the inflammatory infiltrate of CL lesions from patients evolving with good response to therapy ( 2.8 % ± 2.1 % ; median = 2.1 % ; n = 18 ) than poor responders ( 1.3 % ± 1.1 % ; median: 1.05%; n = 6 ; P < 0.05 ). Conclusions. It is the first time that IGF-I was detected in lesions of infectious cutaneous disease, specifically in American tegumentary leishmaniasis. IGF-I was related to chronicity and good response to treatment. We may relate this finding to the efficient anti-inflammatory response and the known action of IGF-I in wound repair. The present data highlight the importance of searching nonspecific factors besides adaptive immune elements in the study of leishmaniasis’ pathogenesis.

BACKGROUND Left ventricular aneurysm (LVA) is indicator of high morbidity in Chagas’ disease. A cross-sectional study performed identified LVA in 18.8% of the chronic chagasic patients (CCP). OBJECTIVE Determine the risk of death of patients with chronic chagasic cardiopathy (CCC) and LVA in 24-year interval. MATERIAL AND METHODS In 1995 a cohort of 298 CCP was evaluated by anamnesis, physical examination, EKG and ECHO and classified in groups: G0 = 86 without cardiopathy; G1 = 156 with cardiopathy without LVA and G2 = 56 with cardiopathy and LVA. 38 patients of G0 and G1 used benznidazole. Information about the deaths was obtained in the notary, death certificates, hospital records and family members. FINDINGS Were registered 113 deaths (37.9%): 107 (35.9%) attributed to cardiopathy and 6 (2.0%) to other causes (p < 0.05). Amongst these 107 deaths, 10 (11.6%) occurred in G0; 49 (31.4%) occurred in G1 and 48 (85.7%) occurred in G2 (p < 0.05). The risk of death was 2.7 and 7.4 times significantly higher in G2, than in G1 and G0, respectively. CONCLUSION Chronic chagasic patients with LVA and ejection fraction < 45% have a higher risk of death than those without.

Serious concerns about the way research is organized collectively are increasingly being raised. They include the escalating costs of research and lower research productivity, low public trust in researchers to report the truth, lack of diversity, poor community engagement, ethical concerns over research practices, and irreproducibility. Open science (OS) collaborations comprise of a subset of open practices including open access publication, open data sharing and the absence of restrictive intellectual property rights with which institutions, firms, governments and communities are experimenting in order to overcome these concerns. We gathered two groups of international representatives from a large variety of stakeholders to construct a toolkit to guide and facilitate data collection about OS and non-OS collaborations. Ultimately, the toolkit will be used to assess and study the impact of OS collaborations on research and innovation. The toolkit contains the following four elements: 1) an annual report form of quantitative data to be completed by OS partnership administrators; 2) a series of semi-structured interview guides of stakeholders; 3) a survey form of participants in OS collaborations; and 4) a set of other quantitative measures best collected by other organizations, such as research foundations and governmental or intergovernmental agencies. We opened our toolkit to community comment and input. We present the resulting toolkit for use by government and philanthropic grantors, institutions, researchers and community organizations with the aim of measuring the implementation and impact of OS partnership across these organizations. We invite these and other stakeholders to not only measure, but to share the resulting data so that social scientists and policy makers can analyse the data across projects.

The golden hamster ( Mesocricetus auratus ) is a susceptible model to Leishmania ( Viannia ) spp.; however, available studies employ different infection protocols, which account for clinical and pathological presentation differences. Herein, L . ( V .) braziliensis preparations were standardized to contain 10 4 , 10 5 , or 10 6 parasites to determine an optimal inoculum that ensured cutaneous lesions without causing a disseminated infection in hamsters. Lesion development was followed for 105 days by size measurements, and skin, draining lymph node, spleen, and sera were investigated to check parasite load, spleen visceralization, cytokine expression, histopathological changes, and anti- Leishmania IgG levels. The lesion emergence time was inversely proportional to the parasite concentration in the inocula. Animals infected by 10 4 parasites presented nodular lesions, while those infected with 10 6 parasites often exhibited ulcerated lesions. The differences in the final lesion sizes were observed between 10 4 and 10 5 inocula or 10 4 and 10 6 inocula. High IFNG expression, anti- Leishmania IgG levels, and parasite load occurred independently of the inoculum used. A mild inflammatory skin involvement was observed in animals infected with 10 4 parasites, while extensive tissue damage and parasite spleen visceralization occurred with 10 5 and 10 6 parasites. These results indicate that inocula with different concentrations of parasites generate differences in the time of lesion emergence, clinical presentation, and systemic commitment, despite high and similar IFNG expression and parasite load. This suggests that a modulation in the immune response to different parasite numbers occurs in an early phase of the infection, which could dictate the establishment and magnitude of the chronic phase of the disease.

Barral Neto, Manoel. Fundacao Oswaldo Cruz. Instituto Goncalo Moniz. Salvador, BA, Brasil. “Documento produzido em parceria ou por autor vinculado a Fiocruz, mas nao consta a informacao no documento”. Centre for Intellectual Property and Policy (CIPP), Faculty of Law, McGill University, Montreal, QC, H3A 1W9, Canada Department of Human Genetics, Faculty of Medicine, McGill University, Montreal, QC, H3A 0C7, Canada Tanenbaum Open Science Institute (TOSI), Montreal Neurological Institute and Hospital, Montreal, QC, H3A 2B4, Canada F1000, London, W1T 4LB, UK Diseases of the Developing World, Global Health R&D, GlaxoSmithKline, Madrid, Spain Fundacao Oswaldo Cruz - Fiocruz, Rio de Janeiro, RJ, 21040-900, Brazil Wellcome Trust, London, NW1 2BE, UK Montreal Neurological Institute and Hospital, Montreal, QC, H3A 2B4, Canada US Department of Agriculture Economic Research Service, Washington, DC, 20024, USA Health Research Board, Dublin, D02 H638, Ireland Arizona State University, Washington, DC, 20006, USA Sage Bionetworks, Seattle, WA, 98121, USA US National Library of Medicine, Bethesda, MD, 20894, USA Research England, UK Research and Innovation, Bristol, BS34 8SR, UK Jisc, Bristol, BS2 0JA, UK Queensland University of Technology, Brisbane, QLD, 4000, Australia The Lens, Canberra, ACT, 2601, Australia African Academy of Sciences, Karen, Nairobi, 00502, Kenya Utrecht University Library, Utrecht, CX, 3584, The Netherlands Structural Genomics Consortium (SGC), University of Oxford, Oxford, OX3 7DQ, UK Data Science Institute, University of Virginia, Charlottesville, VA, 22904, USA University of Cambridge, Cambridge, CB2 3EA, UK Alkermes, Waltham, MA, 02451, USA Department of Psychology, University of Virginia, Charlottesville, VA, 22904-4400, USA Center for Open Science, Charlottesville, VA, 22903-5083, USA Element AI, Montreal, QC, H2W 2R2, Canada GlaxoSmithKline, Stevenage, Herts, SG1 2NY, UK Warwick Research in Nursing, University of Warwick Warwick Research in Nursing, University of Warwick Medical School, Coventry, CV4 7AL, UK University of Wolverhampton, Wolverhampton, WV1 1LY, UK

How should scientific authors choose the best journal to disseminate their research work? If this question was formulated two centuries ago by a biomedical researcher in Europe, the answer would be quite simple: decide first in which language the message should be delivered (e.g., English, German, French, Russian, Latin, Spanish) and then pick one out of the approximately 100 science journals which in that period were publishing papers in one of those languages. The world of scholarly publishing at the beginning of nineteenth century was so small (https://conscicom.org/) that scientists could quickly decide to which journal to send the manuscript.

The diagnosis of visceral leishmaniasis (VL) generally requires the use of invasive tests for the collection of infected tissue (aspirates of bone marrow, spleen, liver or lymph nodes). This difficulty has led to the search for safer and less painful techniques to confirm the occurrence of the disease in children. Polymerase chain reaction (PCR) is a method that is advantageous in that it allows the use of peripheral blood samples for diagnosis. This paper reports the utilisation of PCR on peripheral blood samples to diagnose VL in 45 children in Mato Grosso do Sul, Brazil. This technique is compared with methods carried out using tissue collected by invasive procedures, including direct microscopy, culture and detection of Leishmania DNA by PCR in bone marrow aspirates. The results show that PCR of peripheral blood provides great sensitivity (95.6%) that is similar to that from the PCR of bone marrow aspirates (91.1%) and higher than that achieved with microscopy (80%) or culture (26.7%) methods. PCR of peripheral blood proved to be a suitable tool for the diagnosis of VL in children because it is highly sensitive and safe, with tissue collection being less invasive than in traditional tests.

Introducción. La reacción a la prueba cutánea de Montenegro (a leishmanina) y la lesión de leishmaniasis comparten características fundamentales de la reacción de hipersensibilidad de tipo retardado. Objetivos. Determinar si la respuesta cutánea a leishmanina se aproxima y podría modelar la respuesta inflamatoria e inmune temprana a la infección por Leishmania. Materiales y métodos. Este estudio comparó la respuesta inflamatoria de biopsias de lesiones agudas (tiempo de evolución £1 mes) y lesiones crónicas (tiempo de evolución ³6 meses) con su respectiva reacción a la prueba cutánea de Montenegro, y con la reacción a la prueba de Montenegro de individuos infectados asintomáticos. Resultados. La proporción de linfocitos T CD4+ y CD8+, células fagocíticas mononucleares y granulocitos fue similar entre lesiones agudas y sus reacciones a la prueba cutánea a leishmanina. En contraste, los linfocitos CD4+ (32,6%) representaron una proporción significativamente mas baja, y los linfocitos B (20%) y macrófagos (27%) una proporción significativamente más alta del infiltrado celular en lesiones crónicas que en sus correspondientes reacciones a la prueba cutánea de Montenegro (CD4+: 43,7%, linfocitos B: 0.9%; macrófagos: 17,5%). Se encontró una asociación positiva entre linfocitos T CD8+ y macrófagos ( P=0,038) en la reacción a la prueba cutánea de Montenegro de los individuos con infección asintomática, mientras que los linfocitos T CD8+ y CD4+ estuvieron asociados positivamente en las reacciones a la prueba cutánea de Montenegro de pacientes crónicos ( P=0,002). Adicionalmente, la proporción de linfocitos B en lesiones crónicas (20%) fue más alta que en lesiones agudas (5,3%) ( P=0,002). Conclusión. La reacción cutánea de Montenegro permitió diferenciar la respuesta celular inmune entre infección asintomática y enfermedad crónica, y simula la respuesta temprana a la infección.

Leishmania RNA virus (LRV) has been shown to be a symbiotic component of Leishmania parasites in South America. Nested retro-transcription polymerase chain reaction was employed to investigate LRV1 presence in leishmaniasis lesions from Brazil. In endemic areas of Rio de Janeiro (RJ), no LRV1 infection was observed even with mucosal involvement. LRV1 was only detected in Leishmania (V.) guyanensis cutaneous lesions from the northern region, which were obtained from patients presenting with disease reactivation after clinical cure of their primary lesions. Our results indicated that the severity of leishmaniasis in some areas of RJ, where Leishmania (V.) brazi-liensis is the primary etiological agent, was not associated with Leishmania LRV1 infection.

Se da a conocer la creación del primer programa de integración regional de una red de Institutos de Investigación en Biomedicina pertenecientes a países miembros del MERCOSUR. Se analizan las bases que dieron sustento a su creación y sus objetivos en el mediano y largo plazo. Además, se estima el potencial de los resultados de este programa en los campos de la investigación médica aplicada, educación y biotecnología.It is in our interest, in this brief manuscript, to report the creation of the first program of regional integration of a network of research institutes in Biomedicine belonging to members of the MERCOSUR countries. We discuss some of the foundations that gave sustenance to its creation and its objectives in the medium and long term. In addition, we consider the potential of the results of this program in the fields of applied medical research, education and biotechnology.

Eosinophilic panniculitis may be induced by several triggering factors and this syndrome is described here following the intramuscular injection of antimonial compounds for treatment of American muco-cutaneous leishmaniasis. Three patients, treated with intramuscular injections of pentavalent antimony (Glucantime®) were studied through clinical examination, histopathology and immunologic methods. The three patients developed deep infiltrated plaques at the site of antimony injection. Histopathology disclosed a prominent inflammatory infiltrate of the subcutaneous fat with numerous eosinophils. Immunologic studies could not detect immunoglobulins or complement fractions in the lesions. The final diagnosis was eosinophilic panniculitis occurring as a side-effect of antimonial therapy. The pathogenesis was not clear and it is suggested that the disease could be induced by a physical factor such as pressure or an immunological reaction to antimony.

We have compared the efficacy of two Leishmania (Leishmania) major vaccines, one genetically attenuated (DHFR-TS deficient organisms), the other inactivated [autoclaved promastigotes (ALM) with bacillus Calmete-Guérin (BCG)], in protecting rhesus macaques (Macaca mulatta) against infection with virulent L. (L.) major. Positive antigen-specific recall proliferative response was observed in vaccinees (79% in attenuated parasite-vaccinated monkeys, versus 75% in ALM-plus-BCG-vaccinated animals), although none of these animals exhibited either augmented in vitro gamma interferon (IFN-g) production or positive delayed-type hypersensitivity (DTH) response to the leishmanin skin test prior to the challenge. Following challenge, there were significant differences in blastogenic responses (p < 0.05) between attenuated-vaccinated monkeys and naïve controls. In both vaccinated groups very low levels of antibody were found before challenge, which increased after infective challenge. Protective immunity did not follow vaccination, in that monkeys exhibited skin lesion at the site of challenge in all the groups. The most striking result was the lack of pathogenicity of the attenuated parasite, which persisted in infected animals for up to three months, but were incapable of causing disease under the conditions employed. We concluded that both vaccine protocols used in this study are safe in primates, but require further improvement for vaccine application.

Com o objetivo de estudar o comportamento biológico de cepas do T. cruzi e determinar a possível relação entre o biodema da cepa e as formas clínicas da doença de Chagas, 14 cepas do protozoário isoladas de humanos, procedentes de Pains, Iguatama e Berilo, foram avaliadas. Para estudar o comportamento biológico das cepas, grupos de camundongos albinos suíços, machos, pesando entre 10-15 gramas foram infectados com 1x10(4) tripomastigotas sangüíneos para avaliação da infectividade, parasitemia, morfologia dos tripomastigotas sangüíneos, virulência e distribuição do protozoário nos tecidos. Nove cepas apresentaram comportamento similar ao da cepa São Felipe do biodema II e cinco se caracterizaram como pertencentes ao biodema III. Não foi possível estabelecer uma relação entre o biodema da cepa e gravidade da doença nos pacientes.

Response to treatment with antimonial drugs varies considerably depending on the parasite strain involved, immune status of the patient and clinical form of the disease. Therapeutic regimens with this first line drug have been frequently modified both, in dose and duration of therapy. A regimen of 20 mg/kg/day of pentavalent antimony (Sb5+) during four weeks without an upper limit on the daily dose is currently recommended for mucosal disease ("espundia"). Side-effects with this dose are more marked in elderly patients, more commonly affected by this form of leishmaniasis. According to our experience, leishmaniasis in Rio de Janeiro responds well to antimony and, in cutaneous disease, high cure rates are obtained with 5 mg/kg/day of Sb5+ during 30 to 45-days. In this study a high rate of cure (91.4%) employing this dose was achieved in 36 patients with mild disease in this same geographic region. Side-effects were reduced and no antimony refractoriness was noted with subsequent use of larger dose in patients that failed to respond to initial schedule. A resposta de pacientes com leishmaniose ao tratamento antimonial pode variar em função de fatores como a cepa do parasito envolvido, estado imunológico do paciente e a forma clínica da doença. Os esquemas terapêuticos com antimônio pentavalente (Sb5+) têm sido frequentemente modificados quanto à dose e duração da terapia. Os efeitos colaterais observados com o uso de 20 mg/kg/dia de Sb5+ durante quatro semanas (dose recomendada para o tratamento da forma mucosa) são mais acentuados em pacientes idosos, os mais afetados pela forma mucosa (espundia) da doença. Nossa experiência demonstra que a forma cutânea ocorrendo no Rio de Janeiro responde muito bem ao tratamento com a dose de 5 mg/kg/dia de Sb5+ durante 30 ou 45 dias. Neste estudo foi obtida elevada eficácia (91,4%) com o mesmo regime terapêutico em 36 pacientes com espundia (que evoluem, contudo, com lesão crônica menos destrutiva do que o observado em outras áreas endêmicas) procedentes desta região. Os efeitos colaterais foram reduzidos, não havendo também qualquer resistência ao emprego de doses maiores nos casos que não responderam ao tratamento inicial.

In Brazil, the most common etiological agent of American tegumentary leishmaniasis is Leishmania ( Viannia ) braziliensis . In general, diagnostic techniques envisage the visualization of the parasite, but that technique has a low sensitivity. The main purpose of the present work was to evaluate the PCR as a routine tool for the diagnosis of leishmaniasis. Biopsy specimens from cutaneous or mucosal lesions were taken from 230 individuals from areas where Leishmania is endemic: 216 patients who had a clinical picture suggestive ofl leishmaniasis and 14 individuals with cutaneous lesions due to other causes. Each specimen was processed for histopathologic examination, culture, touch preparation, and DNA isolation. Oligonucleotides that amplify the conserved region of the minicircle molecules of Leishmania were used in a hot-start PCR. While at least one conventional technique was positive for Leishmania for 62% (134 of 216) of the patients, PCR coupled to hybridization was positive for 94% (203 of 216) of the patients. The 14 patients whose clinical picture was not suggestive of leishmaniasis had negative results by all techniques. The impact of the PCR was striking in mucosal disease. While the disease in only 17% (4 of 24) of the patients could be diagnosed by conventional techniques, PCR was positive for 71% (17 of 24) of the patients. Hybridization showed that all cases of disease were caused by parasites belonging to the Viannia subgenus. Altogether, the results indicate that PCR is a valuable tool for the diagnosis of leishmaniasis on a routine basis and is likely to provide valuable epidemiological information about the disease in countries where it is endemic.

Paleoparasitology is the study of parasites found in archaeological material. The development of this field of research began with histological identification of helminth eggs in mummy tissues, analysis of coprolites, and recently through molecular biology. An approach to the history of paleoparasitology is reviewed in this paper, with special reference to the studies of ancient DNA identified in archaeological material.Paleoparasitologia é o estudo de parasitos encontrados em material arqueológico. O desenvolvimento deste campo da pesquisa teve início com a identificação de ovos de helmintos em tecidos mumificados, análise de coprólitos e, recentemente, através da biologia molecular. Neste artigo faz-se uma breve revisão da história da paleoparasitologia com referência especial aos estudos de ADN antigo (ancient DNA) em material arqueológico

Leishmaniasis is an endemic disease in developing countries. The efficacy of therapy is usually evaluated through clinical parameters. To define the parasitologic cure, 20 patients were biopsied before and 1 month to 8 years after treatment. Paraffin-embedded tissue was used for DNA isolation. All patients had a positive polymerase chain reaction (PCR) result before therapy, except 1, for whom no histopathologic material was available. The causative agent was identified as belonging to the Leishmania (Viannia) subgenus by hybridization. Despite clinical healing and absence of reactivation or development of mucosal lesions, PCR was positive in scars of 16 patients (80%). The results suggest that parasites persist in the skin for many years despite treatment. Depending on specific pathogenetic features of the parasite and the immune status of the host, this phenomenon might result in mucosal lesions. Alternatively, it could have a role in the maintenance of immunologic memory in patients living in areas in which leishmaniasis is endemic.

It is in our interest, in this brief manuscript, to report the creation of the first program of regional integration of a network of research institutes in Biomedicine belonging to members of the MERCOSUR countries. We discuss some of the foundations that gave sustenance to its creation and its objectives in the medium and long term. In addition, we consider the potential of the results of this program in the fields of applied medical research, education and biotechnology.

The lack of analytical mechanisms of R&D national networks is a significant problem for policy makers. This paper presents an analytical archetype for performance evaluation of national co-authoring networks based on Social Network Analysis parameters. The model evaluates paper co-authoring data of professionals responsible for culture collections in Brazil and Japan. Were identified professionals from the World Federation for Culture Collections data bank and co-authoring identified from international bibliographic databases. The Brazilian network is the focus of the analysis and the Japanese is the reference network. The Research and Development networks in culture collections are fundamental to the performance of the biotechnological innovation chain in areas such as health, agriculture and industry, and thus fundamental to emerging countries such as Brazil. This country has the fifth largest population in the world, being most of this population user of public health services and it has in the agribusiness one of its main sources of wealth. The model allows a performance evaluation of the networks and identifies improvement paths for the Brazilian network. SNA parameters combined with a reference network showed meaningful elements to the understanding of a R&D networks. This proved helpful for the strategic analysis of the network weaknesses and strengths. The model also proposes policy implications for the three analysis of sub-levels, namely cohesion, subgroups and centralization.

A partir de relato de diagnóstico de leishmaniose visceral em cinco cães dos municípios de Santa Maria, Itaara e Júlio de Castilhos, no centro do Estado do Rio Grande do Sul, foi realizado levantamento sorológico através de teste de imunofluorescência indireta em sangue obtido em papel de filtro de 40 cães de Santa Maria, 20 de Itaara e 11 de Júlio de Castilhos, em geral relacionados aos cinco cães considerados como positivos. Além destes, foi também examinado o sangue de 44 cães vadios de Júlio de Castilhos, 68 de Cachoeira do Sul e 20 de Caçapava do Sul. Soros fracamente reagentes com títulos < 1:40 foram avaliados em ensaios de Ensaio Imuno-enzimático. Amostras de fígado, baço, pulmão e linfonodos de um sexto cão de Santa Maria, considerado como positivo à necropsia, foram examinadas através de histopatologia e PCR. Todos os resultados dos 204 cães foram negativos, indicando que, apesar de não ser impossível a ocorrência de leishmaniose visceral na região, por vários fatores aqui discutidos, ela deve ser pelo menos extremamente rara, sendo necessários estudos mais detalhados do material dos cinco cães tidos como positivos.

Introduction Localized Cutaneous Leishmaniasis (LCL) and Mucosal Leishmaniasis (ML) are two extreme clinical forms of American Tegumentary Leishmaniasis that usually begin as solitary primary cutaneous lesions. Host and parasite factors that influence the progression of LCL to ML are not completely understood. In this manuscript, we compare the gene expression profiles of primary cutaneous lesions from patients who eventually developed ML to those that did not. Methods Using RNA-seq, we analyzed both the human and Leishmania transcriptomes in primary cutaneous lesions. Results Limited number of reads mapping to Leishmania transcripts were obtained. For human transcripts, compared to ML patients, lesions from LCL patients displayed a general multi-polarization of the adaptive immune response and showed up-regulation of genes involved in chemoattraction of innate immune cells and in antigen presentation. We also identified a potential transcriptional signature in the primary lesions that may predict long-term disease outcome. Conclusions We were able to simultaneously sequence both human and Leishmania mRNA transcripts in primary cutaneous leishmaniasis lesions. Our results suggest an intrinsic difference in the immune capacity of LCL and ML patients. The findings correlate the complete cure of L. braziliensis infection with a controlled inflammatory response and a balanced activation of innate and adaptive immunity., Author Summary In Brazil, American tegumentary leishmaniasis is mainly caused by Leishmania braziliensis infection. Usually, it begins as a solitary skin lesion, which is called Localized Cutaneous Leishmaniasis (LCL). However, after this lesion heals, 5% of the patients may develop destructive lesions of the mucosa of nose and throat, which is called Mucosal Leishmaniasis (ML). Currently, there is no technology to identify individuals at risk for ML, and the factors that control the evolution to ML remain unknown. This work aims to study the human gene expression patterns that may contribute to the clinical manifestation of the disease. We used the RNA-Seq technique to study skin lesions from individuals that had LCL (LCL group) and those who developed ML (ML group). Our results suggest that individuals that progressed to ML expressed low levels of genes involved in the immune and inflammatory responses, which might lead to insufficient control of the infection. We were also able to identify a potential gene expression signature to predict long-term disease outcome.

Immunological, parasitological, and molecular techniques were applied to blood samples of dogs to diagnose Leishmania infections. In 1997, 644 domestic dogs were studied. Peripheral blood samples were collected for serological diagnosis and detection of Leishmania parasite by polymerase chain reaction (PCR). The indirect immunofluorescence test was positive in 139 (21.6%) of 644 dogs examined. The PCR was performed in 70 blood samples and 3 bone marrow aspirates. A 120-bp fragment specific for Leishmania was present in PCR hybridization analysis of all seropositive samples in the molecular assays. The PCR hybridization test, which used a minicircle of Leishmania chagasi as a probe, was negative in 20 seronegative dogs. These results suggest that a combined PCR-Southern hybridization technique is a highly sensitive approach to diagnose leishmaniasis in dogs, which are a zoonotic reservoir of leishmaniasis for humans.

Summary Cutaneous lesions caused by Leishmania braziliensis infection occasionally heal spontaneously, but with antimonials therapy heal rapidly in approximately 3 weeks. However, about 15% of the cases require several courses of therapy. Matrix metalloproteinase-2 (MMP-2) and MMP-9 are gelatinases that have been implicated in other chronic cutaneous diseases and skin re-epithelialization. These enzymes are controlled by their natural inhibitors [tissue inhibitors of metalloproteinase (TIMPs)] and by some cytokines. Uncontrolled gelatinase activity may result in intense tissue degradation and, consequently, poorly healing wounds. The present study correlates gelatinase activity to therapeutic failure of cutaneous leishmaniasis (CL) lesions. Our results demonstrate an association between gelatinase activity and increased numbers of cells making interferon (IFN)-γ, interleukin (IL)-10 and transforming growth factor (TGF)-β in lesions from poor responders. Conversely, high levels of MMP-2 mRNA and enhanced MMP-2 : TIMP-2 ratios were associated with a satisfactory response to antimonials treatment. Additionally, high gelatinolytic activity was found in the wound beds, necrotic areas in the dermis and within some granulomatous infiltrates. These results indicate the importance of gelatinase activity in the skin lesions caused by CL. Thus, we hypothesize that the immune response profile may be responsible for the gelatinase activity pattern and may ultimately influence the persistence or cure of CL lesions.

Leprosy presents as a clinical spectrum that is precisely paralleled by a spectrum of immunological reactivity. The disease provides a useful and accessible model, in this case in the skin, in which to study the dynamics of cellular immune responses to an infectious pathogen, including the role of adhesion molecules in those responses. In lesions characterized by strong delayed-type hypersensitivity against Mycobacterium leprae (tuberculoid, reversal reaction, and Mitsuda reaction), the overlying epidermis exhibited pronounced keratinocyte intracellular adhesion molecule 1 (ICAM-1) expression and contained lymphocytes expressing the ICAM-1 ligand, LFA-1. Conversely, in lesions in which delayed-type hypersensitivity was lacking (lepromatous), keratinocyte ICAM-1 expression was low and LFA-1+ lymphocytes were rare. Expression of these adhesion molecules on the cells within the dermal granulomas was equivalent throughout the spectrum of leprosy. The percentage of lymphocytes in these granulomas containing mRNA coding for gamma interferon and tumor necrosis factor alpha, synergistic regulators of ICAM-1 expression, paralleled epidermal ICAM-1 expression. In lesions of erythema nodosum leprosum, a reactional state of lepromatous leprosy thought to be due to immune complex deposition, keratinocyte ICAM-1 expression and gamma interferon mRNA+ cells were both prominent. Antibodies to LFA-1 and ICAM-1 blocked the response of both alpha beta and gamma delta T-cell clones in vitro to mycobacteria. Overall, the expression of adhesion molecules by immunocompetent epidermal cells, as well as the cytokines which regulate such expression, correlates with the outcome of the host response to infection.

Background The oncoproteins of human papillomavirus (HPVs) directly effect cell-cycle control. We hypothesize that regulatory and cell cycle protein expression might be additionally modified in the cervix of HIV/HPV co-infected women. Methods We analyzed the expression of Rb, p27, VEGF and Elf-1 transcriptor factor by immunohistochemistry in 163 paraffin-embeded cervical samples using Tissue Micro-Array (TMA) and correlated this to HIV-1 and HPV infection. Results HIV/HPV co-infection was associated with a significant increase in expression (p < 0.001) of VEGF and p27 in both low and high grade CIN when compared to the cervices of women infected by HPV alone. Decreased Rb expression was evident with increased CIN grade in the cervices of women infected with HPV alone (p = 0.003 average of cells/mm2 in CIN I: 17.9, CIN II/III: 4.8, and tumor 3.9). Rb expression increased 3-fold for both low and high grade CIN with HPV/HIV-1 co-infection compared to HPV infection alone but did not reach statistical significance. There was a significant increase in Elf-1 expression in HPV+/HIV- women with CIN II/III and tumor (average of cells/mm2 in CIN I: 63.8; CIN II/III: 115.7 and tumor: 112.0, p = 0.005), in comparison to controls. Conclusion Co-infection of HPV and HIV leads to significant increase in the VEGF and p27 expression when compared to HPV+/HIV-negative infection that could facilitate viral persistence and invasive tumor development.