Impairment of effector molecules response in diabetes induces susceptibility to Leishmania amazonensis infection.

Type 2 Diabetes is a chronic disease resulting from insulin dysfunction that triggers a low-grade inflammatory state and immune impairment. Leishmaniasis is an infectious disease characterized by chronic inflammation resulted from the parasite's immunomodulation ability. Thus, due to the delicate immune balance required in the combat and resistance to Leishmania infection and the chronic deregulation of the inflammatory response observed in type 2 diabetes, we evaluated the response of PBMC from diabetic patients to in vitro Leishmania amazonensis infection. For that, peripheral blood was collected from 25 diabetic patients and 25 healthy controls matched for age for cells extraction and subsequent experimental infection for 2 or 24 h and analyzed for phagocytic and leishmanicidal capacity by optical microscopy, oxidative stress by GSSG generation, labeling of intracellular mediators by enzyme-Linked immunosorbent assay, and cytokines measurement with cytometric beads array technique. We found that the diabetic group had a higher percentage of infected cells and a greater number of amastigotes per cell. Also, even inducing NF-kB phosphorylation and increasing TNF production after infection, cells from diabetic patients were unable to downregulate NRF2 and generate oxidative stress, which may be associated with the exacerbated levels of IL-6 observed. PBMC of diabetic individuals are more susceptible to infection by L. amazonensis and fail to control the infection over time due to the inability to generate effector microbicidal molecules.

Strongyloides-specific IgA, IgG and IgG immune complex profile in patients with pulmonary tuberculosis.

AIMS: To describe an anti-Strongyloides IgA, IgG and IgG immune complex antibody response profile in patients with pulmonary tuberculosis. METHODS AND RESULTS: Saliva and serum samples were collected from 100 individuals: group I, 50 apparently healthy individuals; and group II, 50 pulmonary tuberculosis patients. The IgA, IgG and IgG immune complex detection were carried out via an ELISA immunoenzymatic test. Optical density medians in saliva samples of IgA antibody (median of 7.21) and IgG-IC (median of 4.95) were significantly higher in tuberculosis group compared to control individuals (median IgA of 3.93 and IgG-IC of 2.38). CONCLUSION: This study presents antibody data to the field of pulmonary tuberculosis and strongyloidiasis coinfection, including saliva samples, and especially IgG immune complex detection.

Evaluation of the Dot-ELISA as a diagnostic test for human strongyloidiasis based on the detection of IgA in saliva.

This study aimed to evaluate the use of saliva samples in the Dot-ELISA test for immunodiagnosis of human strongyloidiasis. The Dot-ELISA presented similar results to the ELISA test, with 70% and 60% sensitivity and 85% and 90% specificity, respectively, for IgA in the saliva. The Dot-ELISA with alternative saliva samples may be a suitable tool for diagnosing human strongyloidiasis, especially in populations with high levels of exposure to helminth.

Temporal and spatial distribution of American tegumentary leishmaniasis in north Paraná: 2010-2015.

INTRODUCTION: Describing the general aspects of American tegumentary leishmaniasis enables the identification of the epidemiological scenario of the disease and the development of preventive actions. METHODS: We analyzed the records of patients with American tegumentary leishmaniasis in north Paraná between 2010 and 2015. RESULTS: We identified 108 cases (mostly in 2014) with the following characteristics: male individuals, rural workers, and ages averaging 56.8 years. Isolated ulcerated lesions were predominant, and Glucantime® was the most frequently used drug. CONCLUSIONS: American tegumentary leishmaniasis remains endemic and affects mostly men found in areas surrounded by woods; its treatment is partially efficient considering its side effects and incidence of recurrences.

Temporal and spatial distribution of American tegumentary leishmaniasis in north Paraná: 2010-2015

Abstract INTRODUCTION: Describing the general aspects of American tegumentary leishmaniasis enables the identification of the epidemiological scenario of the disease and the development of preventive actions. METHODS: We analyzed the records of patients with American tegumentary leishmaniasis in north Paraná between 2010 and 2015. RESULTS: We identified 108 cases (mostly in 2014) with the following characteristics: male individuals, rural workers, and ages averaging 56.8 years. Isolated ulcerated lesions were predominant, and Glucantime® was the most frequently used drug. CONCLUSIONS: American tegumentary leishmaniasis remains endemic and affects mostly men found in areas surrounded by woods; its treatment is partially efficient considering its side effects and incidence of recurrences.

Perfil clínico de pacientes com diagnóstico de tuberculose atendidos no Hospital Universitário de Londrina, Paraná / Clinical profile of patients diagnosed with tuberculosis treated at the University Hospital of Londrina, Paraná

O objetivo deste estudo consistiu em analisar o perfil clínico de pacientes positivos para tuberculose através de um estudo epidemiológico, descritivo e retrospectivo, com base em dados secundários contidos em prontuários dos pacientes atendidos no Hospital Universitário de Londrina compreendendo de janeiro de 2010 a dezembro de 2014. Dos prontuários disponíveis para a análise no momento de estudo 86 casos eram positivos para tuberculose, sendo a maioria do sexo masculino (65/76%) com faixa etária compreendida entre 2 e 91 anos. Os principais sinais clínicos apresentados foram tosse (50/58%), febre (45/52%) e perda de peso (34/40%). Em relação à forma clinica, 58/67% dos pacientes apresentaram a forma pulmonar, e 28/33% a forma não pulmonar. Casos positivos para tuberculose em associação com HIV/AIDS corresponderam a 32/37%. Também foram relatados hábitos prejudiciais dos pacientes nos quais 30/34% eram tabagistas, 20/23% usuários de drogas e 14/16% etilistas. Diante da escassez de dados publicados referentes à tuberculose na nossa região e sua relevância para a saúde pública, nosso estudo contribui com os aspectos epidemiológicos principalmente em relação ao elevado número de casos de coinfecção com Vírus da Imunodeficiência Humana e pacientes com evolução a óbito, auxiliando assim, o desenvolvimento e implementação de campanhas ou projetos que visem o diagnóstico e tratamento precoce.

The objective of this study was to analyze the clinical profile of patients positive for tuberculosis through an epidemiological study, descriptive and retrospective, based on secondary data contained in medical records of patients that attended the Hospital Universitário de Londrina comprising from January 2010 to December 2014. From the medical records available for analysis at the moment of study 86 cases were positive for tuberculosis, being the majority male (65/76%) with age group comprised between 2 to 91 years. The main symptoms presented were cough (50/58%), fever (45/52%) and weight loss (34/40%). Regarding the clinical form, 58/67% of the patients presented the pulmonary form, and 28/33% the non-pulmonary form. Positive cases for tuberculosis in association with HIV/AIDS corresponded to 32/37%. Harmful habits were also reported in patients in which 30/34% were smokers, 20/23% drug users and 14/16% were alcoholics. Against scarcity of published data referring to tuberculosis in our region and its relevance to public health, our study contributes to the epidemiological aspects mainly in relation to the high number of cases of coinfection with Human Immunodeficiency Virus and patients with evolution to death, assisting the development and implementation of campaigns or projects aimed at early diagnosis and treatment.

Brazilian propolis promotes immunomodulation on human cells from American Tegumentar Leishmaniasis patients and healthy donors infected with L. braziliensis.

American Tegumentar Leishmaniasis (ATL) is an infectious disease caused by Leishmania parasites with ineffective treatment. The properties of propolis have been studied in different experimental studies, however, few works have investigated the effects of propolis on human-derived peripheral blood mononuclear cells (PBMC) in leishmaniasis models. Thus, we investigate the immunomodulatory effects of propolis treatment on PBMC from ATL patients and on PBMC from healthy donors infected with Leishmania braziliensis. Our data demonstrate that propolis pretreatment shows immunomodulatory effects on both healthy donors and ATL patients adherent cells, increasing IL-4 and IL-17 and decreasing IL-10, in either the presence or absence of the L. braziliensis infection, demonstrating that propolis contributes with the decrease of the inflammation and could also contribute with parasite control.

Coproparasitological survey of intestinal parasites in the city of Londrina, Parana, Brazil: a retrospective analysis

Intestinal parasites are a major public health problem. It is important to inform and educate the public about these infections, especially where such data are scarce. This study aimed to determine the prevalence of intestinal parasites from the analysis of medical records of individuals of the city of Londrina. We analyzed 11,641 fecal reports from February 2009 to December 2012. Data were cataloged after the completion of parasitological testing of Hoffmann, Pons & Janer, Faust and Kato-Katz. From 11,641 reports, 19.1% were positive for intestinal parasites. Among those, 52.1% pertained to females and 47.9% to males, with predominance of positivity of 27.1% among children 0-10 years. For the regions studied, the northern region stood out with 35.4% of cases and prevalence of 6.8%. Among the pathogenic protozoa, reports of Giardia lamblia comprised 19.1% of positivity, while hookworms were the most frequent among helminths, comprising 7.8% of positive cases. It follows that poor conditions of basic sanitation contribute to the dissemination of these parasites. Early diagnosis is a determinant of successful treatment. Additionally, epidemiological data may be used to study the risk factors for transmission and may result in measures applicable to improving living conditions in the community

Detection of parasite-specific IgG and IgA in paired serum and saliva samples for diagnosis of human strongyloidiasis in northern Paraná state, Brazil.

Human strongyloidiasis is an infection caused by the helminth Strongyloides stercoralis that can be fatal, especially in immunosuppressed patients. The aim of this study is to evaluate parasite-specific IgG and IgA levels using S. venezuelensis third-stage (L3) infective larvae alkaline extract as a heterologous antigen by ELISA in paired serum and saliva samples with improved sensitivity and specificity. Individuals from northern Paraná state, Brazil were divided into three groups: 30 patients copropositive for S. stercoralis (Group I); 30 clinically healthy individuals (Group II); and 30 patients copropositive for other parasites (Group III). The area under ROC curve (AUC), an overall index of diagnostic accuracy, and Kappa index were calculated. Data were analyzed using analysis of variance (ANOVA) followed by a Kruskal-Wallis test. Probability (p) values of <0.05 were regarded as significant. In Group I, IgG was detected in 96.7% serum and in 6.7% saliva samples. IgG was not detected in Group II. In Group III, cross-reactivity was observed for serum IgG in 26.7% and in 6.7% for saliva samples. In Group I, IgA was detected in 76.7% serum and 56.7% saliva samples. In Group II, 3.3% were positive for IgA in serum, whereas IgA was not detected in any saliva samples. Group III showed 6.7% serum and 26.7% saliva-positive samples. The sensitivity values for detection of IgG and IgA in serum samples were 96.7% and 76.7%, respectively. In saliva samples, the sensitivity values for detection of IgG and IgA were 6.7% and 56.7%, respectively. The specificity value was 100% for the detection of IgG in serum and for detection of IgG and IgA in saliva, and 96.7% for detection of IgA in serum samples. The proper choice of immunological diagnosis to supplement parasitological methods is essential to estimate the true prevalence of the parasite, and will permit analysis of population immune response profiles, particularly in northern Paraná state, where there are no previous reports.

Estudo da incidência de parasitas intestinais em verduras comercializadas em feira livre e supermercado de Londrina / Study of the incidence of intestinal parasites in vegetables commercializes in free trade fair and supermarket Londrina

O objetivo deste trabalho foi determinar a presença de parasitas intestinais em amostras de alfaces e almeirões comercializadas em feira livre e supermercado da cidade de Londrina, Paraná. Foram coletadas e analisadas oito amostras de alface (Lactuca sativa) e oito amostras de almeirão (Cichirium intybus). A análise foi realizada no Laboratório de Extensão e Pesquisas em Enteroparasitoses – L.E.P.En. Foram encontrados cistos de Endolimax nana, Entamoeba coli, Entamoeba histolytica, Balantidium coli, larvas de Strongyloides stercoralis, larvas e ovos de Ancilostomídeos e ovos de Ascaris spp. Esses resultados demonstram a necessidade de medidas higiênico-sanitárias, visando à educação da comunidade, possibilitando a melhoria na condição de vida da população.

The objective of this study was to determine the presence of intestinal parasites in samples of lettuce andchicory sold in street market and supermarket Londrina city, Paraná. For this purpose, eight samples of lettuce (Lactuca sativa) and eight samples of chicory (Cichorium intybus) were collected and analyzed. The analysis was performed at the Laboratory for Extension and Research in Enteroparasitosis – L.E.P.En. Cysts of Endolimax nana, Entamoeba coli, Entamoeba histolytica, Balantidium coli and Strongyloides stercoralis larvae, Hookworm larvae and eggs, and eggs of Ascaris spp were found in the samples. These results demonstrate the need for hygienic-sanitary measures, aimed at educating the community, enabling the improvement in living conditions of the population.

Análise dos efeitos da epigalocatequina-3-galato (EGCG) de Camelliasinensis (chá verde) em modelo de hepatotoxicidade químicaexperimental induzida pela Dietilnitrosamina (DEN) / Analysis of the effects of epigallocatechin-3-gallate (EGCG) of Camellia sinensis (green tea) in experimental chemical model of hepatotoxicity induced by Diethylnitrosamine (DEN)

A Dietilnitrosamina (DEN), uma substância reconhecidamente hepatotóxica e carcinogênica, foiutilizada na indução da necrose hepática centrolobular em ratos isogênicos Lewis divididos em 5 grupos de 5 animais. O objetivo deste estudo foi avaliar o efeito quimiopreventivo da epigalocatequina-3-galato (EGCG), de Camellia sinensis (chá verde) no tratamento da hepatotoxicidade celular induzida pela DEN. Foi mensurada a concentração sérica da alanina aminotransferase (ALT) e aspartato aminotransferase(AST) dos diferentes grupos experimentais. No ensaio bioquímico para AST e ALT, houve diferença significativa entre os valores médios do grupo controle (163±70,32) comparado ao grupo DEN (1631±1039,44), sugerindo que a DEN influencia na função hepática. Entretanto, não houve diferença significativa entre o grupo DEN e o tratado com epigalocatequina. A lactato desidrogenase (LDH) éconsiderada um marcador bioquímico comum para avaliação da progressão tumoral, e em relação ao LDH, as amostras não apresentaram diferenças significativas entre o grupo DEN (1385,5±43,13) e DEN + EGCG 150mg ou DEN + EGCG 200mg 1537,5±1010,45). Neste trabalho foi demonstrado que a epigalocatequina nas concentrações de 150 e 200 mg/Kg não induziram alterações hepáticas e também não foi possível verificar nenhuma quimioproteção pela EGCG em animais inicialmente tratados comDEN durante 24 horas. Sendo assim, novos experimentos com diferentes concentrações de EGCG sãonecessários para comprovar seu possível efeito quimioprotetor.

Diethylnitrosamine (DEN), a known hepatotoxic and carcinogenic substance, was used in the induction of centrilobular hepatic necrosis in isogenic Lewis rats divided into 5 groups with 5 animals. The aim of this study was to evaluate the chemopreventive effect of epigallocatechin-3-gallate (EGCG)from Camellia sinensis (green tea) in the treatment of cellular hepatotoxicity induced by DEN. It was measured the serum concentration of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) of the different experimental groups. In the biochemistry assay for AST and ALT, there was significant difference between median values of control group (163±70.32) compared to DEN group(1631±1039.44), suggesting that DEN influences on hepatic function. However, there was no significant difference between DEN group to that treated with epigallocatechin. Lactate dehydrogenase (LDH) is considered a common biochemical marker for evaluation of tumor progression, and regarding LDH,the samples presented no significant differences between the DEN group (1385.5±43.13) and DEN + EGCG 150mg or DEN + EGCG 200mg (1537.5± 1010.45). In this work it was demonstrated that epigallocatechin concentrations of 150 and 200 mg/kg did not induce liver alterations and though was not verified any chemoprotective effect by EGCG in animals initially treated with DEN for 24 hours. Moreover, new experiments with different concentrations of EGCG are needed to verify its possiblechemoprotector effect.

Detection of Lsr2 gene of Mycobacterium leprae in nasal mucus

In the present study, nasal mucus from patients with leprosy were analyzed by PCR using specific primers for Lsr2 gene of Mycobacterium leprae. The presence of Lsr2 gene in the nasal mucus was detected in 25.80% of patients with paucibacillari leprosy, and 23.07% of contacts. Despite the absence of clinical features in the contact individuals, it was possible to detect the presence of Lsr2 gene in the nasal mucus of these individuals. Therefore, PCR detection of M. leprae targeting Lsr2 gene using nasal mucus samples could contribute to early diagnosis of leprosy.

Concanavalin-A induces IL-17 production during the course of Candida albicans infection.

In a previous study, our group verified that 100% of mice survived to a lethal dose of Candida albicans following pretreatment with concanavalin-A (Con-A) for 3 days. This work proposed to investigate whether treatment could mediate an adaptative immune response involving T(H) 17 cells. A significant increase in IL-17 levels at 6 h postinfection was observed and was maintained up to 18 h in the Con-A group, whereas in control mice, a reduction in this cytokine was verified. In addition, T(H) 17 cells develop in the presence of TGF-ß, IL-1 ß, and IL-6 that were increased significantly 2 h postinfection in Con-A-treated mice. Macrophages were involved in the process, engulfing greater numbers of yeast cells, and were activated through TNF-α and interferon-γ produced at significant levels at 2 h postinfection. A significant increase in IL-12 levels was also observed at 2 h postinfection. Thus, activated macrophages were probably more capable of killing and processing Candida antigens, signalizing an adaptative immune response. Macrophages from controls did not prevent yeast-to-hyphae transition and were partially destroyed, as shown in scanning microscopy. These results suggest that treatment with Con-A facilitated the triggering of T(H) 17 and T(H) 1 responses via IL-17 and IFN-γ production, leading to the resolution of C. albicans infection.

Artin M enhances TNF-α production and phagocytosis of Candida albicans mediated by dectin-1 and mannose receptors.

The activities of dectin-1 and mannose receptors on phagocytosis of Candida albicans and the production of TNF-α by macrophages from mice pretreated for 3 days with extract of Artocarpus intergrifolia seeds (jack extract), Artin M or jacalin were studied. Macrophages from these mice were coincubated with C. albicans CR15 (yeast), in the presence of mannose (50mM) plus mannan (100 µg) or laminarin (1mg). Phagocytosis was significantly enhanced to 52% in macrophages from mice pretreated intraperitoneally for 3 days with jack extract (500 µg/250 µl PBS). Reduction in phagocytosis from 52% to 34% (P<0.05) occurred in the presence of mannose receptor inhibitors and from 52% to 16% (P<0.01) in the presence of dectin-1 inhibitor laminarin, whereas only 20% of control macrophages phagocytosed blastoconidia. Similar results were verified for pretreatment of mice with Artin M (2.5 µg/250 µl PBS), but not for jacalin (25 µg/250 µl PBS). Macrophages from mice pretreated 3 days previously with jack extract or Artin M and then coincubated for 2h with C. albicans presented a significant increase in TNF-α production, correlating with significantly less transition of yeast to filamentous forms compared to pretreatment with jacalin. These results suggest that Artin M, but not jacalin present in jack extract significantly increased TNF-α production and the activity of mannose and dectin-1 receptors.

Protective effect of Artin M from extract of Artocarpus integrifolia seeds by Th1 and Th17 immune response on the course of infection by Candida albicans.

The immunoregulatory effect of Artin M and jacalin from extract of Artocarpus integrifolia seeds (jack extract) against infection with Candida albicans was investigated. Swiss mice received jack extract containing 500 µg protein/ml PBS intraperitoneally (i.p.) or PBS alone and after 72 h were infected i.p. with C. albicans CR15 (10(7)) and sacrificed after 30 min, 2, 6, 24, and 72 h. ELISA analysis revealed that in jack extract-treated mice IFN-γ was predominantly produced versus IL-10 in control mice. These results suggest that jack extract induced a protective immune response, since C. albicans clearance was complete at 72 h postinfection. Jack extract presents two lectins (Artin M and jacalin) with distinct biological properties. Artin M was able to induce IL-12 production by macrophages. Also, Artin M in different concentrations, associated with jacalin or in jack extract induced both IFN-γ and IL-17 production. As a consequence, phagocytic and candidacidal activity increased significantly. Alanine aminotransferase activity (ALT) was used as parameter for damage of the liver. The activity of ALT correlated with inoculum size that increased significantly in control group, however, mice pretreated with jack extract 3 days before infection presented normal ALT. Mice pretreated with jack extract that received a lethal inoculum of Candida presented 90% survival versus 20% among controls or mice pretreated with jacalin. Thus, the results suggest that Artin M by itself, associated with jacalin or present in jack extract is able to induce protective Th1 and Th17 immune responses against Candida albicans infection.

Increased tumour necrosis factor-alpha production, higher mannose receptor activity and ability to kill Candida by concanavalin-A-activated macrophages.

In a previous study, our group verified that mice pretreated with concanavalin-A (Con-A) produced more tumour necrosis factor (TNF)-alpha and presented greater Candida clearance from the peritoneal cavity, liver and spleen, which yielded a higher survival rate than control animals. In this work, the hypothesis that macrophages were of crucial importance in overcoming the infection was tested. Thus, peritoneal macrophages from mice pretreated for 3 days with Con-A or phosphate-buffered saline (PBS) were coincubated with CR1, CR15 and 577 isolates of Candida albicans for 0.5, 1 and 2 h. The ability of Con-activated macrophages to produce TNF-alpha, ingest via mannose receptors and kill all the isolates was significantly greater compared with PBS-treated macrophages, and activated macrophages exhibited a lower incidence of apoptosis, verified by binding to annexin V-fluorescein isothiocyanate. The transition of yeast cells to filamentous forms during coincubation for 2 h with control macrophages was about 73-80%, whereas in the presence of Con-A-activated macrophages, it was 35-40%. Our results suggest that a greater clearance of C. albicans infection through treatment with Con-A is probably due to the activation of macrophages, which produce more TNF-alpha, express more mannose receptors and are better endowed to kill ingested C. albicans.