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Disseminated cutaneous leishmaniasis (DCL) is an uncommon form of Leishmania braziliensis infection. It remains unknown why some people develop this clinical condition. We describe 14 DCL patients in Northeast Brazil during 2015-2018. These patients regularly drank large amounts of alcohol, possibly increasing their risk for DCL.

Leishmania braziliensis is the main causative agent of American tegumentary leishmaniasis in Brazil. Current treatment includes different drugs that have important side effects and identification of cases of parasite resistance to treatment support the search for new therapeutic strategies. Recent findings have indicated that CXCL10, a chemokine that recruits and activates Th1 cells, NK cells, macrophages, dendritic cells and B lymphocytes, is a potential alternative to treat Leishmania infection. Here, we tested CXCL10 immunotherapy against experimental infection caused by an antimony-resistant isolate of Leishmania braziliensis. Following infection, mice were treated with CXCL10 for 7 days after onset of lesions. We demonstrate that mice treated with CXCL10 controlled lesion progression and parasite burden more efficiently comparing to controls. An increased IFN-γ, IL-10, TGF-β and low IL-4 production combined with a distinct inflammatory infiltrate composed by activated macrophages, lymphocytes and granulomas was observed in the CXCL10-treated group comparing to controls. However, CXCL10 and Glucantime combined therapy did not improve CXCL10-induced protective effect. Our findings reinforce the potential of CXCL10 immunotherapy as an alternative treatment against infection caused by L. braziliensis resistant to conventional chemotherapy.

Sensory receptors are distributed throughout the oral cavity, pharynx, and larynx. Laryngeal sensitivity is crucial for maintaining safe swallowing, thus avoiding silent aspiration. Morphologic description of different receptor types present in larynx vary because of the study of many different species, from mouse to humans. The most commonly sensory structures described in laryngeal mucosa are free nerve endings, taste buds, muscle spindles, glomerular and corpuscular receptors. This study aimed at describing the morphology and the distribution of nerve endings in premature newborn glottic region. Transversal serial frozen sections of the whole vocal folds of three newborns were analyzed using an immuno-histochemical process with a pan-neuronal marker anti-protein gene product 9.5 (PGP 9.5). Imaging was done using a confocal laser microscope. Nerve fiber density in vocal cord was calculated using panoramic images in software Morphometric Analysis System v1.0. Some sensory structures, i.e. glomerular endings and intraepithelial free nerve endings were found in the vocal cord mucosa. Muscle spindles, complex nerve endings (Meissner-like, spherical, rectangular and growing) spiral-wharves nerve structures were identified in larynx intrinsic muscles. Nervous total mean density in vocal cord was similar in the three newborns, although they had different gestational age. The mean nerve fiber density was higher in the posterior region than anterior region of vocal cord. The present results demonstrate the occurrence of different morphotypes of sensory corpuscles and nerve endings premature newborn glottic region and provide information on their sensory systems.

American visceral leishmaniasis is caused by the protozoan Leishmania infantum. The control of the disease depends on the magnitude of the Th1 cell response and IL-10 producing regulatory T cells. Administration of chemokine, such as CXCL10, has shown promising results in the leishmaniasis treatment. Previous studies from our group have shown that CXCL10 induces a reduction in parasite burden in the spleen and a decrease in IL-10 and TGF-β production in L. infantum-infected BALB/c mice. This work investigated whether CXCL10-treatment reduces IL-10 + Treg cell populations (CD4

INTRODUCTION: The dengue classification proposed by the World Health Organization (WHO) in 2009 is considered more sensitive than the classification proposed by the WHO in 1997. However, no study has assessed the ability of the WHO 2009 classification to identify dengue deaths among autopsied individuals suspected of having dengue. In the present study, we evaluated the ability of the WHO 2009 classification to identify dengue deaths among autopsied individuals suspected of having dengue in Northeast Brazil, where the disease is endemic. METHODS: This retrospective study included 121 autopsied individuals suspected of having dengue in Northeast Brazil during the epidemics of 2011 and 2012. All the autopsied individuals included in this study were confirmed to have dengue based on the findings of laboratory examinations. RESULTS: The median age of the autopsied individuals was 34 years (range, 1 month to 93 years), and 54.5% of the individuals were males. According to the WHO 1997 classification, 9.1% (11/121) of the cases were classified as dengue hemorrhagic fever (DHF) and 3.3% (4/121) as dengue shock syndrome. The remaining 87.6% (106/121) of the cases were classified as dengue with complications. According to the 2009 classification, 100% (121/121) of the cases were classified as severe dengue. The absence of plasma leakage (58.5%) and platelet counts

American visceral leishmaniasis is caused by the protozoan Leishmania infantum. The control of the disease depends on the magnitude of the Th1 cell response and IL-10 producing regulatory T cells. Administration of chemokine, such as CXCL10, has shown promising results in the leishmaniasis treatment. Previous studies from our group have shown that CXCL10 induces a reduction in parasite burden in the spleen and a decrease in IL-10 and TGF-β production in L. infantum-infected BALB/c mice. This work investigated whether CXCL10-treatment reduces IL-10 + Treg cell populations (CD4+CD25+Foxp3+ and Tr1) and induces morphological changes in the spleen. BALB/c mice were infected and treated or not with CXCL10 on the 1st, 3rd and 7th days of infection. CXCL10-treatment was able to reduce the parasite load in the spleen in L. infantum-infected BALB/c mice and this decrease in the number of parasites correlated with the decrease in size of this organ in treated animals compared to untreated animals. 7, 23, and 45 days post-treatment (p.t.), the phenotype and frequency of IL-10 + Treg cells were evaluated by flow cytometry, and the morphological changes of the spleen were analyzed by optical microscopy. After 7 and 23 days p.t., CXCL10-treated animals showed a significant reduction of CD25-Foxp3−IL-10+ (Tr1) cells in the spleen when compared to untreated animals, whereas CD4+CD25+Foxp3+IL-10+ Treg cells reduced later at 23rd and 45th days p.t. Furthermore, while untreated animals showed a significant positive correlation between IL-10 production and Tr1 cells, in CXCL10-treated group this correlation was negative. Thus, these findings show that treatment with CXCL10 chemokine in L. infantum-infected BALB/c mice results in suppression of IL10+ Treg (Foxp3+ and Tr1) cells in the spleen, associated with a reduction in parasite load and splenomegaly.

Federal University of Cear?. Fortaleza, CE, Brazil. Federal University of Cear?. Fortaleza, CE, Brazil. Federal University of Cear?. Fortaleza, CE, Brazil / Servi?o de Verifica??o de ?bitos. Fortaleza, CE, Brazil. Cear? State Central Public Health Laboratory. Fortaleza, CE, Brazil. Federal University of Cear?. Fortaleza, CE, Brazil / Servi?o de Verifica??o de ?bitos. Fortaleza, CE, Brazil. Federal University of Cear?. Fortaleza, CE, Brazil / Servi?o de Verifica??o de ?bitos. Fortaleza, CE, Brazil. University of Fortaleza. Fortaleza, CE, Brazil / Cear? State Secretariat of Health. Fortaleza, CE, Brazil. Cear? State Central Public Health Laboratory. Fortaleza, CE, Brazil. Federal University of Cear?. Fortaleza, CE, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. University of Virginia School of Medicine. Charlottesville, VA, USA. Chan School of Public Health. Harvard T.H. Boston, MA, USA / University of California, San Francisco. CA, USA. Federal University of Cear?. Fortaleza, CE, Brazil. Postmortem examination of 7 neonates with congenital Zika virus infection in Brazil revealed microcephaly, ventriculomegaly, dystrophic calcifications, and severe cortical neuronal depletion in all and arthrogryposis in 6. Other findings were leptomeningeal and brain parenchymal inflammation and pulmonary hypoplasia and lymphocytic infiltration in liver and lungs. Findings confirmed virus neurotropism and multiple organ infection.

OLIVEIRA, Augusto Cesar Aragao ; FONTES, Raissa Matos ; PRACIANO, Claudenia Costa ; ARAUJO, Fernanda Montenegro de Carvalho ; CAVALCANTI, Luciano Pamplona de Goes ; COLARES, Jeova Keny Baima ; POMPEU, Margarida Maria de Lima ; LIMA, Danielle Malta. Recognition of leptospirosis in dengue-suspected cases during dengue outbreak in Ceara State, Brazil. African Journal of Microbiology Research, v. 8, n. 17, p. 1789-1792, abr. 2014.

Recruitment of a specific cell population after Leishmania infection can influence the outcome of the disease. Cellular migration in response to Leishmania or vector saliva has been reported in air pouch model, however, cellular migration induced by Leishmania associated with host's blood and vector saliva in this model has not been described. Herein we investigated cellular migration into air pouch of hamster after stimulation with combination of L. chagasi and host's blood and Lutzomyia longipalpis saliva. Migration induced by saliva was 3-fold more than those induced by L. chagasi alone. Additionally, L. chagasi associated with blood and saliva induced significantly even more leukocytes into air pouch than Leishmania alone. L. chagasi recruited a diverse cell population; however, most of these cells seem to have not migrated to the inflammatory exudate, remaining in the pouch lining tissue. These results indicate that L. chagasi can reduce leukocyte accumulation to the initial site of infection, and when associated with vector saliva in the presence of blood components, increase the influx of more neutrophils than macrophages, suggesting that the parasite has developed a strategy to minimize the initial inflammatory response, allowing an unlimited progression within the host. This work reinforces the importance of studies on the salivary components of sand fly vectors of leishmaniasis in the transmission process and the establishment of the infection.

Leishmaniasis is a vector-borne disease transmitted by phlebotomine sand fly. Susceptibility and refractoriness toLeishmaniadepend on the outcome of multiple interactions that take place within the sand fly gut. Promastigote attachment to sand fly midgut epithelium is essential to avoid being excreted together with the digested blood meal. Promastigote and gut sand fly surface glycans are important ligands in this attachment. The purpose of the present study was to evaluate the interaction of three lectins isolated from leguminous seeds (Diocleinae subtribe), D-glucose and D-mannose-binding, with glycans onLutzomyia migoneimidgut. To study this interaction the lectins were labeled with FITC and a fluorescence assay was performed. The results showed that onlyDioclea violacealectin (DVL) was able to interact with midgut glycans, unlikeCratylia floribundalectin (CFL) andCanavalia gladiatalectin (CGL). Furthermore, when DVL was blocked with D-mannose the interaction was inhibited. Differences of spatial arrangement of residues and volume of carbohydrate recognition domain (CRD) may be the cause of the fine specificity of DVL for glycans in the surface onLu. migoneimidgut. The findings in this study showed the presence of glycans in the midgut with glucose/mannose residues in its composition and these residues may be important in interaction betweenLu. migoneimidgut andLeishmania.

Protein-free guar gum (DGG) was oxidized (DGGOX) or sulfated (DGGSU) by insertion of new groups in C-6 (manose) and C-6 (galactose), for DGGOX and DGGSU, respectively. Rats were subjected to anterior cruciate ligament transection (ACLT) of the knee, joint pain recorded using the articular incapacitation test, and the analgesic effect of intraarticular 100μg DGG, DGGOX or DGGSU solutions at days 4-7 was evaluated. Other groups received DGG or saline weekly, from days 7 to 70 and joint damage assessed using histology and biochemistry as the chondroitin sulfate (CS) content of cartilage. The molar mass of CS samples was obtained by comparing their relative electrophoretic mobility to standard CS. DGG but not DGGOX or DGGSU significantly inhibited joint pain. DGG significantly reversed the increase in CS, its reduced electrophoretic mobility, and histological changes following ACLT, as compared to vehicle. Structural integrity accounts for DGG benefits in experimental osteoarthritis.

Dengue remains a problem in Brazil, and a substantial number of cases that progress to death are not diagnosed by health services. We evaluated the impact of a protocol adopted by the Coroner's Office Rocha Furtado (CO-RF) for the detection of unreported deaths from dengue in Brazil. We evaluated prospectively cases of deaths referred to the CO-RF with suspicion of dengue and those referred with other diagnosis in which the pathologists suspected dengue as the cause of death. Biological material was collected from all bodies autopsied, for which the suspected cause of death was dengue, between January 2011 and December 2012. Of the 214 bodies autopsied, 134 (62.6%) tested positive for dengue; of these cases, 121 were classified as dengue according to the World Health Organization's case definition (1997 or 2009, as appropriate). Thus, CO-RF detected 90 deaths from dengue, which were not suspected during disease progression. This CO-RF protocol, through a combined effort of the surveillance and laboratory teams, increased the detection of fatal dengue cases by 5-fold. This is the largest series of autopsies performed in cases of death related to dengue in the world to date.

Em algumas áreas endêmicas de leishmaniose tegumentar(LT) causada por Leishmania (Viannia) braziliensis existem evidências de transmissão domiciliar, e de que cães infectados estariam associados a casos humanos de LT. Neste trabalho, são apresentados os resultados de um estudo realizado na Serra de Baturité, no Ceará, em 1993, quando foram investigadas as evidências de transmissão domiciliar e a associação entre LT humana e infecção canina. Casos humanos de LT foram indivíduos que apresentavam úlceras de bordas elevadas, arredondadas, de diâmetro > 5 mm, duração > 15 dias e uma Reação de Montenegro positiva. Cães soropositivos foram aqueles que apresentavam uma prova de imunofluorescência indireta positiva. A idade foi dividida em dois estratos, de 1 a 10 e 11 a 89 anos. A incidência de LT não estava associada ao sexo (Razão de Incidência (RI) = 1,21; IC 95%: 0,86 - 1,72) e nem à idade (RI = 1,38; IC 95%: 0,97 - 1,96). Para os indivíduos que conviviam com um ou mais cães soropositivos, a incidência de LT humana foi 45% mais elevada (RI = 1,45; IC 95%: 1,02 - 2,06) depois de ajustar para o sexo, e 48% (RI = 1,48%; IC 95%: 1,04 - 2,10) depois de ajustar para idade. Considerando-se o risco domiciliar, a incidência de LT humana foi duas vezes mais elevada (RI = 2,01; IC 95%:1,32 - 3,06) nos domicílios que possuíam um ou mais cães soropositivos. Conclui-se que os indivíduos foram acometidos de LT independente do sexo e da idade e que os casos humanos estavam associados à presença de cães infectados. Portanto, estes resultados suportam a hipótese de transmissão domiciliar da leishmaniose tegumentar na Serra de Baturité, no Ceará.

Over the last few years, several cases of feline leishmaniasis (FL) with cutaneous and visceral forms have been reported around the world. Nonetheless, the real susceptibility of cats to infection with Leishmania spp. and the outcome of leishmaniasis in these animals are poorly understood. Experimental studies on feline models will contribute to the knowledge of natural FL. Thus, in order to determine the susceptibility of domestic cats (Felis catus) to experimental infection with Leishmania braziliensis, 13 stray cats were infected with 10(7) promastigotes by the intradermal route in the ear and nose simultaneously and followed up for 72 weeks. Soon after infection, the earliest indication of a lesion was a papule on the ear at 2 weeks post-infection (w.p.i.). The emergence of satellite papules around the primary lesion was observed about 4 w.p.i. Two weeks later these papules coalesced and formed a huge and irregular nodule. Thereafter, there was lesion dissemination to the external and marginal surface of the ipsilateral ear, and later to the contralateral ear. At 10 w.p.i., some nodules became ulcerated. Nose lesions presented a similar evolution. At both sites, the largest lesion sizes occurred at 10 w.p.i. and started to decrease 15 days later. Ear and nose nodules healed at 32 and 40 w.p.i., respectively. Specific L. braziliensis IgG antibody titers (optical density> or = 0.01 as positive result) were detected as early as 2 w.p.i. (0.09 +/- 0.02) in only three animals (23%), and all cats had positive titers at 20 w.p.i. (0.34 +/- 0.06). Only three animals (38%) continued to show positive serology at 72 w.p.i. (0.08 +/- 0.02). Up to that time, none of the cats had lesion recurrence. In a feline model of cutaneous leishmaniasis, it seems that there is no correlation between active lesions and positive serology. The implications of these data are discussed.

Inflammatory events duringLeishmania braziliensisinfection in mice were investigated. Large lesions were directly correlated with the inflammatory reaction but not with parasite burden. DifferentL. braziliensisstrains induce different paces of chemokine expression patterns, leading to diverse cell recruitment and differential inflammatory responses.

To the Editor: Leishmania infantum is endemic to northeastern Brazil. It is responsible for visceral leishmaniasis (VL), a major emerging health problem in urban areas. Transmission occurs predominantly by the Lutzomyia longipalpis sand fly, but transfusion-associated VL, caused by L. infantum, has been reported from southern Europe and, by L. donovani, on the Indian subcontinent (1,2). Most L. infantum infections are asymptomatic (3), raising concern that the parasite could be present in donated blood from otherwise healthy residents in areas to which it is endemic (4). VL caused by L. infantum is endemic to 20 of Brazil’s 27 states; an annual average of 3,553 cases occur nationwide, with 54% of all cases reported from Brazil’s northeastern region. The state of Ceara historically ranks first or second in number of cases; an annual average of 467 cases were reported during the last decade (5). Thirty-eight percent of cases were reported from Fortaleza, the capital, where 28.4% of the state’s population resides. Over a 10-year-period, 277 (7.8%) persons with VL have died, and 109 (39%) of VL-related deaths have occurred in Fortaleza. Sixty-nine percent of blood donors for Ceara reside in Fortaleza. To determine the prevalence of Leishmania infection among healthy blood donors, we tested blood donated to the State of Ceara Public Blood Bank. Compulsory serologic testing was also done for Trypanosoma cruzi (Chagas disease), hepatitis B and C, Treponema pallidum (syphilis), human T-cell lymphotropic virus types 1 and 2, and HIV-1 and -2. In the blood bank, 60% of units are centrifuged to separate the buffy coat in preparation for platelet separation. During May–November 2011, we randomly selected 431 buffy coats and tested them for Leishmania spp. by ELISA and PCR. To separate plasma from cells, 10 mL of buffy coat was centrifuged in Ficoll-Hypaque (Histopaque −1077, Sigma-Aldrich, Sao Paulo, Brazil). We tested plasma for leishmanial IgG by ELISA using a modified protocol of Evans et al. (6). IOC/L2906 L. infantum strain (MHOM/BR/2002/LPC-RPV) was used as the source of promastigote antigens. In addition, DNA was extracted from the mononuclear cell preparation, and PCR was performed with primers 150 (5′-GGG[G/T]AGGGGCGTTCT[G/C]CGAA-3′) and 152 (5′-[G/C][G/C][G/C][A/T]CTAT[A/T]TTACACCAACCCC-3′) that target the 120-bp conserved region of the Leishmania kDNA minicircle present in all Leishmania spp (7). As a positive control, kDNA was extracted from L. amazonensis promastigotes, strain BA-125 (MHOM/BR/87), characterized by PCR and isoenzymes (8). All PCR-positive samples were purified and sent to Ludwig Biotec (Alvorada, Brazil) for sequencing by ACTGENE-Molecular Analysis. The Federal University of Ceara Ethics Committee approved this study. Buffy coats from 57 (13.2%) serum samples from 431 donors were positive for leishmanial IgG, and 20 (4.6%) were positive for Leishmania spp. DNA. Sequencing of all PCR-positive samples confirmed the Leishmania genus. Three donors tested positive by both ELISA and PCR. Overall, the prevalence of leishmanial infection was 17.1% of blood donors. Eighty of the 431 units tested positive for >1 of compulsorily screened infections and were rejected. Of the remaining 351 that were negative for co-infection, 43 (12.2%) were positive for leishmanial IgG and 15 (4.3%) for Leishmania spp. DNA. Two donors were positive for both by ELISA and PCR. The prevalence of Leishmania infection among blood units accepted for transfusion was 16%. The results demonstrate a surprisingly high prevalence of Leishmania infection in blood donors in Fortaleza, several times higher than that other diseases for which blood is screened (Figure). In a recent study in Salvador, Brazil (9), 5.4% of blood donors had leishmanial antibodies, of which 68% were positive by its PCR targeting kDNA amplification. Figure Comparison of the prevalence of Leishmania infantum as tested by PCR and ELISA and of other infections compulsorily tested in 431 blood donors in Fortaleza, state of Ceara, northeastern Brazil. HBV, hepatitis B virus; HCV, hepatitis C virus; HTLV, ... The percentages of antibody- or PCR-positive units capable of transmitting Leishmania and the outcomes are unknown. Viable Leishmania might not be in the blood of all PCR-positive donors, and even when present, the inoculum might be reduced by removal of infected circulating mononuclear phagocytes in the buffy coat, or parasites might be affected by steps involved in preparation or storage. However, if we consider units that test positive by PCR as being potentially infectious, the number of recipients at risk is of substantial concern. For example, in 2011, there were 99,933 blood donations to the State of Ceara Public Blood Bank. After compulsory screening for the other bloodborne pathogens, 93,238 units were accepted for transfusion. Extrapolating from the PCR-positive rate of 4.3%, a total of 4,009 recipients possibly were exposed to infection. Further studies are needed to determine whether recipients of blood from donors who are PCR positive and/or leishmanial antibody positive become infected with L. infantum. Persons with advanced AIDS or other immunosuppressive conditions seemingly would be at greatest risk for VL. In Brazil, legislation requires that all blood for transfusion be tested for T. cruzi ., hepatitis B and C, T. pallidum, human T-cell lymphotropic virus types 1 and 2, and HIV-1 and -2. As additional information becomes available, screening for L. infantum also might be advisable to reduce the possibility of the recipient becoming infected, developing VL, and possibly being a reservoir of infection in the community (10), particularly in Ceara and other regions where the prevalence of L. infantum infection is high.

A 52-year-old male farm worker from Ceara state, northeast Brazil, presented with a 4-month history of skin lesions. Lesions appeared first on the face and legs, than spread across much of his body, including the genitalia, soles of feet, and scalp. He reported daily fevers for the first 2 weeks of illness. On examination the patient had 178 lesions of different sizes and aspects1–3; ulcerated, crusted (Figure 1A and ​andBB ), papular, nodular, and pustular. Many had apparent bacterial superinfections (Figure 1C). The remainder of the physical examination was normal. A complete blood count and liver and kidney function tests were within normal limits. Serological tests for syphilis and human immunodeficiency virus were negative. A skin biopsy imprint revealed amastigotes. Skin culture in Novy-MacNeal-Nicolle medium grew promastigotes characterized as Leishmania (Viannia) braziliensis by isoenzymes. The leishmanin skin test was positive. Bacterial superinfection of the ulcers was treated with cephalexin. Because of a contraindication to treatment with pentavalent antimony (individual > 50 years of age), and lack of medical infrastructure where he lived for providing amphotericin B, the patient was treated with oral fluconazole, 9 mg/kg per day.4 Over the following 6-month period of treatment, the lesions healed completely, many leaving no scars (Figure 1D). Disseminated cutaneous leishmaniasis is an uncommon presentation of Leishmania (V.) braziliensis infection. Most of the lesions are thought to be due to bloodstream dissemination, not to multiple sand fly bites. The reason that some people develop disseminated cutaneous leishmaniasis is not fully understood.3 Figure 1. (A and B) More than 20 crusted and ulcerated lesions can be seen on the face, with involvement of the forehead, eyelid, nose, cheeks as well as the upper lip. (C) The larger ulcerated lesion is clearly draining purulent secretion. (D) Patient's face after ...

Introduction : Immunoglobulins, soluble antigens, cells, cytokines and other immune system products can be transferred from infected mother to her offspring, leading to suppression or stimulation of immune response. Objective : To evaluate the influence of gender and maternal infection with Leishmania braziliensis in the course of the disease in the offspring of hamsters. Meodths: Offspring born from infected mother (IMO) or non-infected mother (NIMO) by Leishmania braziliensis, both sexes, was infected with the same strain of the mother after 30 days of life and followed for 18 weeks. We evaluated the thickness of the lesion, parasite load and histology of the lesions. Results : The number of parasite in both lesions and lymph node of IMO offspring showed a significant reduction in the 5th week post-infection compared to the NIMO offspring; however, this did not correspond to clinical symptoms. Histopathological analysis revealed that in the IMO offspring, the inflammatory process was more prominent. In relation to gender, it was observed that the male offspring showed lesion thickness and higher parasite burden than females. Conclusion : Maternal infection by L. braziliensis in hamsters does not appear to influence the course of the disease in the homologous offspring infection, as well as the male offspring presented augmented susceptibility to L. braziliensis infection regardless of whether they were born from IMO or NIMO. Also, the reduction of the granuloma index in the IMO offspring, together with the higher inflammatory response, suggests a less effective cellular response in the chronic phase of the disease in these animals.

We report for the first time the successful use of fluconazole to treat cutaneous leishmaniasis due to Leishmania braziliensis. We used escalating doses from 5 to 8 mg/kg per day. At a dose of 5 mg/kg per day, 75% patients were cured, and at 8 mg/kg per day, the cure rate was 100%. Fluconazole was well tolerated.

The effects of 1,8-cineole on d-galactosamine/lipopolysaccharide (GalN/LPS)-induced shock model of liver injury was investigated in mice. The co-administration of GalN (700 mg kg−1, i.p.) and LPS (5 μg kg−1, i.p.) greatly elevated serum concentrations of tumour necrosis factor-α (TNF-α), alanine aminotransferase and aspartate aminotransferase, and induced massive hepatic necrosis and lethality in 100% of control mice. Pretreatment with 1,8-cineole (400 mg kg−1, p.o.) and dexamethasone (1 mg kg−1, s.c.),60 min before GalN/LPS, offered complete protection (100%) against the lethal shock and acute elevation in serum TNF-α and serum transaminases. Hepatic necrosis induced by GalN/LPS was also greatly reduced by both 1,8-cineole and dexamethasone treatment. The results indicate that 1,8-cineole protects mice against GalN/LPS-induced liver injury through the inhibition of TNF-α production, and suggest that 1,8-cineole may be a promising agent to combat septic-shock-associated pathologies.

This study aims to evaluate the in vitro and in vivo leishmanicidal activity of lapachol, a naphthoquinone found in the seeds and heartwood of certain tropical plants, and to compare its efficacy with a reference drug, sodium stibogluconate (Pentostam®). These compounds (0.0125–4.0 mg/mL) were evaluated in vitro against intracellular amastigotes of Leishmania (Viannia) braziliensis (LVb), then tested in an animal model (hamster) to try to reproduce the leishmanicidal activity. In vitro, lapachol exhibited an anti-amastigote effect, whereas in vivo it did not prevent the development of LVb-induced lesions at an oral dose of 300 mg/kg/day for 42 days. Pentostam® demonstrated a significant anti-amastigote effect in vitro for LVb and apparent clinical cure in vivo (60 mg/kg/day). However, it could not completely eradicate parasites from the tissues of infected animals. The observation that lapachol exerts leishmanicidal activity in vitro without offering significant protection against LVb-infected lesions in hamsters suggests that lapachol in vivo might possibly inhibit the microbicidal functioning of macrophages. Alternatively, it might be transformed into an inactive metabolite(s) or neutralized, losing its leishmanicidal activity. It is also possible that an optimal and sustained plasma level of the drug could not be achieved at the dose used in this study. Copyright © 2001 John Wiley & Sons, Ltd.

Objective and Design: We studied the contribution of periarthritis and synovitis to gait disturbance in zymosan (Zy)-induced arthritis.¶Methods: Sixty Wistar rats were subjected to injection of Zy (1mg) into their right knee joints. A first group of animals (GI) had Zy injected through the intact skin. A second group (GII) had Zy injected directly into the articular cavity after excision of the skin and subcutaneous tissue surrounding the joint. Gait disturbance was evaluated using the rat-knee joint incapacitation test. Increase in vascular permeability and cell influx were assessed in joint fluids and joint histology was performed.¶Results: Zy injection induced a dose-dependent gait disturbance which was maximal at the third/fourth hour of arthritis, being significantly greater in GI rats, whereas cell influx (neutrophils ≤80%) was maximal at the sixth hour. Cell influx and increase in vasopermeability did not differ between both groups. Histology revealed no significant difference between GI and GII. A third group (GIII), subjected to immune-complex arthritis, that received anti-bovine serum albumin (BSA) antibodies intra-articularly and BSA i.v., did not present gait disturbance, despite the increase in cell counts.¶Conclusions: Vascular permeability increase and cell influx are phenomena independent of gait disturbance. Neutrophils do not seem to contribute to development of gait disturbance in Zy arthritis. Sensitization of specific pain receptors in periarticular rather than in synovial tissue is responsible for gait disturbance in Zy-induced arthritis.

O programa de controle do calazar, adotado pela Fundação Nacional de Saúde (FNS), não tem conseguido reduzir a níveis aceitáveis a incidência de calazar humano. Utiliza como método diagnóstico a imunofluorescência de eluato de sangue e sacrifíca os cães infectados com uma média de 80 dias após a coleta. A longa permanência do cão infectado na área e a baixa sensibilidade do teste utilizado podem ser importantes para esta falha. Neste trabalho, compara-se o programa de rotina da FNS, com outra estratégia baseada na identificação de cães infectados pelo enzyme-linked immunosorbent assay (ELISA) e eliminação dos cães infectados dentro de um prazo máximo de 7 dias. A prevalência do calazar canino foi medida nas duas áreas, antes e 10 meses após a medição inicial. Na área submetida ao controle de rotina da FNS observou-se um decréscimo de 9% na prevalência, enquanto na submetida ao método proposto a redução observada foi de 27%, sendo esta diferença sinificativamente maior (p = 0,0015).The kala-azar control program, adopted by the Fundação Nacional de Saúde- FNS (National Health Foundation) has not been able to reduce to an acceptable level the incidence of human cases. The diagnostic method utilized is a blood eluate immunofluorescence. A dogs diagnosed as infected is eliminated a mean of eighty days after the blood collection. The low sensitivity of the test used and the continuing residence of the infected dog in the region due to the elimination delay may be critical in the lack of success of this program. In this study, the FNS standard canine control method is compared to a strategy based on ELISA identification of infected dog and elimination within 7 days. In both study areas the canine seroprevalence was noted ten months before and ten months after the intervention. In the routine FNS area a 9% decrease in seroprevalence was noted, compared to statistically significant greater 27%, reduction (p = 0.0015) in the ELISA intervention area.

Background: The repair of the immature squamous epithelium following HPV infection may mimic HSIL in adolescent women. Aim: to study the utility of p16 INK4a and Ki-67 in diagnosis of cervical squamous lesions in adolescents and young adults. Materials and Methods: In a cross-sectional study, the evaluation of p16 INK4a and Ki-67 immunohistochemistry was performed on 72 cervical biopsies of adolescents and young adults women diagnosed as negative for malignancy and intraepithelial lesion (NML) (n = 18) or positive for low grade (LSIL) (n = 31) and high grade (HSIL) (n = 23) squamous intraepithelial lesions in two references services in Fortaleza-Brazil. Data was evaluated using Fisher′s test and Kappa index. Results: p16 INK4a was positive in 81% of HSIL, 19% of LSIL and in no NML (P < 0.0001). Ki-67 was positive in 74%, 32% and 5.5% of HSIL, LSIL and NML, respectively. p16 INK4a and Ki-67 in the diagnosis of HSIL showed high sensitivity and negative predictive value. Kappa index was very good for p16 INK4a (k = 0.72). Conclusions: In adolescents and young adults p16 INK4a alone or with Ki-67 represents important tool to reduce mistaken diagnosis of HSIL and to avoid overtreatment.

Background Intra-articular (i.a) injection of a purified Guar gum (GG) solution or hydrogel provided analgesia similar to that of Hylan G-F20 in experimental osteoarthritis (eOA) suggesting that rheology is not crucial to viscosupplements. Objectives Evaluate the effect of purified GG (DGG), sulfated (DGGSU) of oxidized (DGGOX) GG on pain and cartilage damage in eOA Methods GG was oxidated with a n-oxil-2, 2, 6, 6-tetra metilpiperidina (TEMPO) reagent or subjected to sulfation to produce carboxylated and sulphate groups. Rats subjected to anterior cruciate ligament transection (ACLT) of the knee received 100 μg GG, sulfated or oxidized GG or vehicle (NT) i.a. at day 4 to 7 after ACLT. Joint pain was recorded using the articular incapacitation test. A group received GG weekly or vehicle from day 7 to 70 and joint damage assessed by histology and biochemically, measuring hondroitin-sulfate (CS) cartilage content (μg/mg) and Mw of CS through PAGE. Results FT-InfraRed spectra of DDGSU and DDGOX were different from protein free DGG. DGGSU has a new intense band at 1262 cm –1 , due to S=O. Spectral difference occurs in DGGOX but not DGG and a C=O stretching is present in DGGOX spectrum (COOH group). 13 C NMR spectrum of DGGOX and DGG revealed absence of the signal at 63.7 ppm present in DGG and two new signals at 177.5 and 177.8 ppm. Absence of the signal from C-6 of free mannose indicates it was substituted in the oxidation. The new signals are from C=O group from COOH. 1 H- 13 C HSQC spectra exhibited a shift to a lower d value of the proton H-1 from mannose residue in DGGOX, indicating substitution at C-6 from mannose. New signals at 70.3, 75.6, and 100.5 ppm were observed in 13 C NMR spectrum of sulfated gum due to the insertion of sulfate group in the primary carbon (C-6). All derivatives show lower molar mass than the original guar gum. DGG M w =3.9 x 10 6 g/mol, DGGOX and DGGSU, 2.6 x 10 6 and 2.8 x 10 6 g/mol, respectively. DGGOX has the lowest intrinsic (2.2 dL/g) and apparent viscosity (9.0 mPa s).DGG exhibits the highest values ([h] =6.2 dL/g and h app =58 mPa s). Intermediate values are obtained for DGGSU ([h] =4.9 dL/g and h app =39 mPa s). GG significantly reduced joint pain, as compared to vehicle whereas oxidat ion or sulfation of GG abrogated GG analgesia. GG also prevented the increase in both CS content (43.8±4.7 μg/mg) and Mw (mobility: 1.18 ±0.03), as compared to NT (p Conclusions The structural integrity of GG is crucial to its analgesia. Intra-articular polysaccharides can be analgesic and chondroprotective regardless of being water-soluble. Acknowledgement Supported by grants 302218/2014–9 and 459334/2014–0 from CNPq-Brasil). Disclosure of Interest None declared

1. Curso de Medicina, Universidade de Fortaleza, Fortaleza, CE. 2. Departamento de Patologia e Medicina Legal, Universidade Federal do Ceara, Fortaleza, CE. 3. Centro de Pesquisa em Virologia, Faculdade de Medicina, Universidade de Sao Paulo, Ribeirao Preto, SP. 4. Setor de Virologia, Laboratorio Central de Saude Publica do Estado do Ceara, Fortaleza, CE. 5. Nucleo de Vigilância Epidemiologica, Secretaria Estadual de Saude do Estado do Ceara, Fortaleza, CE. 6. Departamento de Saude Publica, Universidade Federal do Ceara, Fortaleza, CE.

Patients from across the spectrum of clinical manifestations of Leishmania chagasi infection were evaluated for in vitro correlates of immunity. Peripheral blood mononuclear cells were assayed for parasite-specific lymphoproliferation, cytokine generation, and the capacity to activate autologous macrophages to kill intracellular amastigotes. Patients with acute kala-azar were generally unreactive in each of these assays. Children with subclinical infection demonstrated relatively low levels of proliferation and interferon-gamma production, but none went on to develop overt kala-azar during the study. Patients evaluated after therapy for kala-azar demonstrated yet higher levels of lymphoproliferation and cytokine generation and produced low but significant levels of cytokines in vitro in response to parasite antigens, but not during the activation of infected macrophages. Finally, peripheral blood mononuclear cells from adults with positive delayed-type hypersensitivity responses and no history of kala-azar showed the broadest reactivity in vitro. These patients' cells generated the largest amounts of activating cytokines in vitro during the activation of autologous macrophages to a leishmanicidal state.

Evidence that combined glucosamine sulfate and chondroitin sulfate (Gluchon) or isolated glucosamine (Glu) modifies joint damage in osteoarthritis (OA) is still lacking. We studied joint pain and cartilage damage using the anterior cruciate ligament transection (ACLT) model. Wistar rats were subjected to ACLT of the right knee (OA) or sham operation. Groups received either Glu (500 mg/kg), Gluchon (500 mg/kg glucosamine +400 mg/kg chondroitin) or vehicle (non-treated—NT) per os starting 7 days prior to ACLT until sacrifice at 70 days. Joint pain was evaluated daily using the rat-knee joint articular incapacitation test. Structural joint damage was assessed using histology and biochemistry as the chondroitin sulfate (CS) content of cartilage by densitometry (microgram per milligram dried cartilage), comparing to standard CS. The molar weight (Mw) of the CS samples, used as a qualitative biochemical parameter, was obtained by comparing their relative mobility on a polyacrylamide gel electrophoresis to standard CS. Gluchon, but not Glu, significantly reduced joint pain (P

We investigated the effect of an extract from a helminth (Ascaris suum) in zymosan-induced arthritis (ZYA) or collagen-induced arthritis (CIA). Rats and mice, respectively, received 1 mg and 0.1 mg zymosan intra-articularly (i.a.). Test groups received anA. suumextract either per os (p.o.) or intraperitoneally (i.p.) 30 min prior to i.a. zymosan. Controls received saline. Hypernociception was measured using the articular incapacitation test. Cell influx, nitrite, and cytokine levels were assessed in joint exudates. The synovia and distal femoral extremities were used for histopathology. Cartilage damage was assessed through determining glycosaminoglycan (GAG) content. DBA/1J mice were subjected to CIA. The test group receivedA. suumextract i.p. 1 day after CIA became clinically detectable. Clinical severity and hypernociception were assessed daily. Neutrophil influx was determined using myeloperoxidase activity. TheA. suumextract, either i.p. or p.o., significantly and dose-dependently inhibited cell influx and hypernociception in ZYA in addition to reducing GAG loss and ameliorating synovitis. TheA. suumextract reduced i.a. levels of NO, interleukin-1β (IL-1β), and IL-10 but not tumor necrosis factor alpha (TNF-α) in rats subjected to ZYA while reducing i.a. IL-10, but not IL-1β or TNF-α, levels in mice. Clinically, mice subjected to CIA treated with theA. suumextract had less severe arthritis. Hypernociception, myeloperoxidase activity, and synovitis severity were significantly reduced. These data show that a helminth extract given p.o. protects from arthritis severity in two classical arthritis models. ThisA. suumeffect is species independent and functions orally and parenterally. The results show clinical and structural benefits whenA. suumextract is given either prophylactically or therapeutically.

A 55-year-old physician had a PPD made as part of a clinical evaluation. Laboratory tests were unremarkable and PPD, read in 48 hours, showed an area of induration of 18 mm and an erythema of 51 mm (Panel A). A 2 mm punch biopsy was done in the center of the induration 96 hours after the injection. Histopathology showed an intense perivascular cuffing with macrophages and lymphocytes and edema involving the dermis. The macrophages were predominant and most of them showed cellular swelling. There was some degree of vasculitis, as shown (Panel C). A positive PPD test is generally defined as greater than 10 mm of (Panel B) induration, not erythema, and can be measured by using a medium-point ballpoint pen to draw a line starting 1 to 2 cm away from the skin reaction (induration) and moving toward its center. The pen is lifted when resistance is felt. The procedure is repeated from the opposite direction and the distance between opposing lines is measured (Panel A). Virtually all people with PPD greater than 15 mm of induration are infected with Mycobacterium tuberculosis (Mtb), so the physician is infected with Mtb. PPD is also called Tuberculin and Mantoux test after the French physician Charles Mantoux (1877-1947). In 1934 Florence B. Seibert (1897-1991), an American Scientist, developed the method to obtain the Purified Protein Derivative-PPD, that became the basis for the standard Tuberculosis Test. Seibert, was a Professor of Biochemistry at the Henry Phipps Institute at the University of Pennsylvania.

To the Editor—We appreciate Drs Torres and Suarez's interest in our article, “High-Dose Oral Fluconazole Therapy Effective for Cutaneous Leishmaniasis Due to Leishmania (Vianna) braziliensis” [1]. They refer to evidence of Leishmania amazonensis isolated from human as well as domestic and wild reservoir hosts from the state of Ceara and suggest it is an important etiologic agent of cutaneous leishmaniasis in the region. Concerning human hosts, their statement is based on 2 publications [2, 3] that showed a single isolate of L. amazonensis from only 1 human case of cutaneous leishmaniasis. Both articles deal with the same parasite grown from a human skin biopsy sample in 1984. Because that event was so rare, the question as to whether it was an imported case remains. In respect to domestic and wild reservoirs of L. amazonensis in the state of Ceara, there is no proof of isolates of L. amazonensis from any reservoir or from sandflies. The data available for isolates from humans [4–7], from domestic and wild reservoir hosts [4, 6], and from sandflies [4, 6, 8, 9] support our statement: Leishmania (Viannia) braziliensisis is the only known agent of cutaneous leishmaniasis in the state of Ceara. In a large study of 272 human isolates from 39 different municipalities, all were characterized as L. braziliensis [5]. In another report [6], the author studied 354 isolates from humans, rodents, dogs, and sandflies; all were characterized as L. braziliensis. Furthermore, because the patients in our study were from several different endemic areas, the hypothesis that the response of our patients was due to aspecific clinical syndrome caused by L. braziliensis from a specific endemic area is not true. The lymphadenopathy described in L. braziliensis infection [5] has been documented in other regions of Brazil, and its detection depends on the moment patients seek medical attention and on a comprehensive physical examination because this lymphadenopathy is an early manifestation of infection and, as a rule, the lymphadenopathy is painless and abates as the ulcer progresses. There is evidence that 1 strain of L. braziliensis from 1 region may be genetically different from another strain from other regions [10], and it is possible that these strains may respond differently to therapy. Hence, it is critically important that fluconazole be tested in other areas where L. braziliensis and other species of Leishmania are endemic, keeping in mind that optimal weight-based dosing is very likely to be an important determinant of therapeutic success [1].

In vivo administration of Canavalia brasiliensis lectin (at the time of infection, or maintained throughout the infection) reduced the lesions of highly susceptible BALB/c mice infected by Leishmania amazonensis. At the doses used C. brasiliensis lectin (ConBr) does not interfere with penetration or fate of Leishmania in the macrophages in vitro. Since Interferon-gamma (IFN-gamma) is the major macrophage activating factor, and considered a critical element in the successful immune response against leishmaniasis, we explored its participation in this phenomenon. ConBr either in vivo or in vitro induced IFN-gamma production in normal or in leishmania-infected BALB/c mice. However we were unable to change the course of disease by in vivo IFN-gamma administration (although IFN-gamma preparations were effective in inducing a leishmanicidal effect in vitro on L. amazonensis-infected peritoneal macrophages). Additionally, IFN-gamma neutralization with anti-IFN-gamma monoclonal antibody did not alter the protection conferred by ConBr administration. These data show that lectin administration in vivo is protective in the otherwise unchecked L. amazonensis infection of BALB/c mice, and suggest that such effect is not mediated by IFN-gamma.

Enlarged regional lymph nodes have been reported to accompany the cutaneous lesions of Leishmania (Viannia) braziliensis (= L. braziliensis). A survey in Ceara State, Brazil indicated that 77% of persons (456 of 595) with parasitologically confirmed cutaneous leishmaniasis reported lymphadenopathy in addition to skin lesions. A group of 169 persons with recently diagnosed leishmaniasis and lymph nodes measuring ≥ 2 cm in diameter (mean = 3.6 cm, maximum = 10.5 cm) underwent detailed clinical examination. Lymphadenopathy preceded the skin lesions in more than two-thirds of these, on the average by two weeks. Cultures of lymph node aspirates yielded Leishmania more frequently (86%) than cultures of aspirates of skin (53%) or biopsies of skin (74%). Parasites were isolated from the peripheral blood of one patient. Persons with lymphadenopathy gave a history of fever and had enlarged livers or spleens more often than a comparison group of 50 persons with cutaneous lesions but no lymphadenopathy. Persons with lymphadenopathy had more intense leishmanin skin reactions and lymphocyte proliferation following stimulation with specific antigens, whereas persons without lymphadenopathy had a higher frequency of previous infection. Isolates of parasites from both groups were identified as L. braziliensis. These data demonstrate the early spread of L. braziliensis beyond the skin and suggest differences in host immunity between persons with and without lymphadenopathy. Leishmaniasis braziliensis should be considered in cases of unexplained lymphadenopathy in endemic areas.

RESUMO Objetivo Relatar a experiência da aplicação de material didático composto por catálogo com imagens macroscópicas e microscópicas, disponível em meio impresso e digital, acompanhado de coleções de lâminas, instrumento criado por alunos do mestrado e iniciação científica, focando a autonomia do aprendizado em Patologia Humana para os alunos da graduação. Métodos Foi elaborado um questionário para ser aplicado aos alunos, para avaliação de resultados alcançados pelo modelo criado, que será oferecido aos estudantes de diversos cursos que possuem a disciplina de Patologia Humana, oferecida na Faculdade de Medicina da Universidade Federal do Ceará (UFC). O catálogo relata a história e evolução da doença de uma paciente com tuberculose sistêmica, suas manifestações clínicas, exames físicos e laboratoriais, além de achados de necropsia, com diferentes aspectos da resposta inflamatória. Resultados Os alunos que viveram esta experiência se manifestaram a favor da implantação da nova ferramenta como material didático da disciplina e sua manutenção nos laboratórios. Conclusão A inovação nas aulas é necessária, e este modelo surge como um material complementar para a melhoria e motivação das aulas práticas no tema proposto.

The initial encounter of Leishmania with its host's immune system is important in the outcome of infection. Previous studies have shown that PBMCs from healthy volunteers (HV) exposed to Leishmania differ in IFN-γ production. We have expanded such observations evaluating the profile and kinetics of cytokines (IFN-γ, IL-12p70, IL-10, IL-13), chemokines (CCL5, CCL3, CCL4, CXCL10), and chemokine receptors (CCR1,CCR5, CXCR3, CCR4) in vitro L. amazonensis-stimulated of HV́s PBMCs. HVs were divided in groups of high (HR) or low (LR) IFN-γ responders. In both groups, HR and LR, after L. amazonensis infection there was a predominance of IL-10 and IL-13 over IFN-γ production, while IL-12 was produced in similar amount. Regarding chemokines, a more striking difference was observed for CCL3 expression that was lower at 12 hours and 48 hours post infection in LR than in HR. Interestingly, a downregulation of CCR5 and a greater expression of CCR4 were found in low IFN-γ responders. These data suggest that early after L. amazonensis infection there is a cytokine milieu dominated by IL-13 and IL-10, and despite of this environment, IFN-γ is produced, supporting the complexity of the response. It is noteworthy that the pattern of immune response is mounted in first hours after Leishmania stimulation, with the definition of the differentiation of Th1 versus Th2 cells. It remains to be determined if such an in vitro difference has an in vivo counterpart in terms of susceptibility to infection. Keywords: Leishmania amazonensis, interleukin-10, interleukin-13

Background Dengue is an important worldwide public health problem, and continues to spread in Brazil. This article presents the first Brazilian case report of the death of an indigenous child by dengue fever. Case presentation In August 2013, a child aged 2 years and from the Tremembé ethnic group, who was previously healthy with no complaints, suddenly presented intense crying, precordial pain, and general malaise. A few minutes after these non-specific symptoms, the patient started tonic–clonic convulsions and had cyanosis, a substantial increase in body temperature to the touch, cold sudoresis, sphincter relaxation, and unconsciousness. This situation remained for 15 minutes, progressing to respiratory insufficiency, with consequent absence of peripheral pulses. Death was confirmed approximately 40 minutes after the first symptoms. An autopsy was performed using the usual techniques. Immunohistochemistry was positive for dengue, and microscopic examination indicated micro perivascular edema and cerebral hemorrhage. Conclusion Considering that the death occurred during the major endemic seasonal period for dengue fever, primary clinical evidence suggestive of viral infection presenting with sudden and quick death, and positive immunohistochemistry results, the case was closed as severe dengue fever. Clinicians must consider dengue as a diagnostic hypothesis among the indigenous population in Brazil.