Blockade of TLR2 and TLR4 Attenuates Inflammatory Response and Parasite Load in Cutaneous Leishmaniasis.

Human cutaneous leishmaniasis (CL) caused by Leishmania braziliensis is characterized by a pronounced inflammatory response associated with ulcer development. Monocytes/macrophages, the main cells harboring parasites, are largely responsible for parasite control. Toll-like receptor (TLR) signaling leads to the transcription of inflammatory mediators, such as IL-1ß and TNF during innate immune response. TLR antagonists have been used in the treatment of inflammatory disease. The neutralization of these receptors may attenuate an exacerbated inflammatory response. We evaluated the ability of TLR2 and TLR4 antagonists to modulate host immune response in L. braziliensis-infected monocytes and cells from CL patient skin lesions. Following TLR2 and TLR4 neutralization, decreased numbers of infected cells and internalized parasites were detected in CL patient monocytes. In addition, reductions in oxidative burst, IL-1ß, TNF and CXCL9 production were observed. TNF production by cells from CL lesions also decreased after TLR2 and TLR4 neutralization. The attenuation of host inflammatory response after neutralizing these receptors suggests the potential of TLR antagonists as immunomodulators in association with antimonial therapy in human cutaneous leishmaniasis.

The application of rapid prototyping in facial surgeries: a series of case studies / Aplicação de prototipagem rápida em cirurgias de face: uma série de estudos de casos

Aim: the aim of this paper is to report on the applicability of biomodels obtained from computerized tomography (CT) images through the technique of stereolithography (SLA) and three-dimensional printing (3DP), in the clinical case reports of patients who underwent surgeries involving dentoskeletal deformity, oral pathology and oral and maxillofacial trauma. Methodology: clinical case 01 deals with fractures in the orbital and zygomatic arch region that required reconstruction and correction of diplopia, by means of CT images, using the technique of three-dimensional printing through SLA, generating a mirrored biomodel for surgical planning and the making of customized prosthesis. In clinical case 02, by means of CT images, a biomodel utilizing the 3DP technique showed the total area invaded by invasive ameloblastoma, making it possible to plan the osteotomy with maximum preservation of the adjacent tissue and prior modeling of the plate. In clinical case 03, rapid prototyping technology (RP) was used to make customized prosthesis for temporomandibular joint, with the goal of correcting a serious idiopathic pathology provoking the resorption of the right and left condyles. Discussion: complex cases require the team to have recourse to technology for the implementation of the procedures, in order to offer excellent quality treatment to the patient, in addition to facilitating surgical planning and permitting the construction of customized prostheses. Conclusion: the rapid prototyping for the acquisition of biomodels is an important auxiliary tool for the surgical team.

Objetivo: o objetivo deste estudo foi informar sobre a aplicabilidade de biomodelos obtidos a partir de imagens de tomografia computadorizada (TC) através da técnica de estereolitografia (SLA) e impressão tridimensional (3DP), nos casos clínicos de pacientes submetidos a cirurgias envolvendo deformidade dentoesquelética, patologia oral e trauma oral e maxilofacial. Metodologia: o caso clínico 01 trata de fraturas na região do arco orbital e zigomático que requerem reconstrução e correção da diplopia, por meio de imagens de TC, usando a técnica de impressão tridimensional através de SLA, gerando um biomodelo espelhado para planejamento cirúrgico e fabricação de próteses personalizadas. No caso clínico 02, por meio de imagens TC, um biomodelo utilizando a técnica 3DP mostrou a área total invadida pelo ameloblastoma, possibilitando o planejamento da osteotomia com preservação máxima do tecido adjacente e modelagem prévia da placa. No caso clínico 03, a tecnologia de prototipagem rápida (RP) foi utilizada para fabricar próteses personalizadas para a articulação temporomandibular, com o objetivo de corrigir uma séria patologia idiopática que provocou a reabsorção dos côndilos direito e esquerdo. Discussão: casos complexos exigem que a equipe recorra à tecnologia para realização dos procedimentos, a fim de oferecer um tratamento de excelência e qualidade ao paciente, além de facilitar o planejamento cirúrgico e permitir a construção de próteses personalizadas. Conclusão: a prototipagem rápida para aquisição de biomodelos é uma importante ferramenta auxiliar para equipe cirúrgica.

Leishmania braziliensis Infection Enhances Toll-Like Receptors 2 and 4 Expression and Triggers TNF-α and IL-10 Production in Human Cutaneous Leishmaniasis.

Cutaneous leishmaniasis (CL) caused by infection with Leishmania braziliensis is characterized by an exaggerated inflammatory response that controls the parasite burden, but also contributes to pathology. While myeloid cells are required to eliminate the parasite, recent studies indicate that they may also participate in the inflammatory response driving disease progression. The innate immune response to leishmania is driven in part by the Toll-like receptors (TLRs) TLR2, TLR4, and TLR9. In this study, we used flow cytometric analysis to compare TLR2 and TLR4 expression in monocyte subsets (classical, intermediate, and non-classical) from CL patients and healthy subjects (HS). We also determined if there was an association of either the pro-inflammatory cytokine TNF or the anti-inflammatory cytokine IL-10 with TLR2 or TLR4 expression levels after L. braziliensis infection. In vitro infection with L. braziliensis caused CL monocytes to up-regulate TLR2 and TLR4 expression. We also found that intermediate monocytes expressed the highest levels of TLR2 and TLR4 and that infected monocytes produced more TNF and IL-10 than uninfected monocytes. Finally, while classical and intermediate monocytes were mainly responsible for TNF production, classical monocytes were the main source of IL-10. Collectively, our studies revealed that up-regulated TLR2/4 expression and TNF production by intermediate/inflammatory subsets of monocytes from patients correlates with detrimental outcome of cutaneous leishmaniasis.

Characterization of Neutrophil Function in Human Cutaneous Leishmaniasis Caused by Leishmania braziliensis.

Infection with different Leishmania spp. protozoa can lead to a variety of clinical syndromes associated in many cases with inflammatory responses in the skin. Although macrophages harbor the majority of parasites throughout chronic infection, neutrophils are the first inflammatory cells to migrate to the site of infection. Whether neutrophils promote parasite clearance or exacerbate disease in murine models varies depending on the susceptible or resistant status of the host. Based on the hypothesis that neutrophils contribute to a systemic inflammatory state in humans with symptomatic L. braziliensis infection, we evaluated the phenotype of neutrophils from patients with cutaneous leishmaniasis (CL) during the course of L. braziliensis infection. After in vitro infection with L. braziliensis, CL patient neutrophils produced more reactive oxygen species (ROS) and higher levels of CXCL8 and CXCL9, chemokines associated with recruitment of neutrophils and Th1-type cells, than neutrophils from control healthy subjects (HS). Despite this, CL patient and HS neutrophils were equally capable of phagocytosis of L. braziliensis. There was no difference between the degree of activation of neutrophils from CL versus healthy subjects, assessed by CD66b and CD62L expression using flow cytometry. Of interest, these studies revealed that both parasite-infected and bystander neutrophils became activated during incubation with L. braziliensis. The enhanced ROS and chemokine production in neutrophils from CL patients reverted to baseline after treatment of disease. These data suggest that the circulating neutrophils during CL are not necessarily more microbicidal, but they have a more pro-inflammatory profile after parasite restimulation than neutrophils from healthy subjects.

Immunologic Markers of Protection in Leishmania (Viannia) braziliensis Infection: A 5-Year Cohort Study.

BACKGROUND: The control of Leishmania braziliensis by individuals with subclinical infection (SC) are unknown. METHODS: A cohort of 308 household contacts (HCs) of patients with cutaneous leishmaniasis (CL) was established in 2010 in an endemic area and followed up for 5 years. Whole-blood cultures stimulated with soluble Leishmania antigen and a Leishmania skin test (LST) were performed in years 0, 2, and 4. The identification of the lymphocyte subsets secreting interferon (IFN) γ and the ability of monocytes to control Leishmania were determined. RESULTS: During follow-up, 118 subjects (38.3%) had evidence of L. braziliensis infection. Of the HCs, CL was documented in 45 (14.6%), 101 (32.8%) had SC infection, and 162 (52.6%) did not have evidence of exposure to L. braziliensis The ratio of infection to disease was 3.2:1. IFN-γ production, mainly by natural killer cells, was associated with protection, and a positive LST result did not prevent development of disease. Moreover, monocytes from subjects with SC infection were less permissive to parasite penetration and had a greater ability to control L. braziliensis than cells from patients with CL. CONCLUSIONS: Protection against CL was associated with IFN-γ production, negative LST results, impaired ability of Leishmania to penetrate monocytes, and increased ability to control Leishmania growth.

The Role of Nitric Oxide and Reactive Oxygen Species in the Killing of Leishmania braziliensis by Monocytes from Patients with Cutaneous Leishmaniasis.

Human cutaneous leishmaniasis (CL) caused by Leishmania braziliensis, presents an exaggerated Th1 response that is associated with ulcer development. Macrophages are the primary cells infected by Leishmania parasites and both reactive oxygen species (ROS) and nitric oxide (NO) are important in the control of Leishmania by these cells. The mechanism involved in the killing of L. braziliensis is not well established. In this study, we evaluate the role of ROS and NO in the control of L. braziliensis infection by monocytes from CL patients. After in vitro infection with L. braziliensis, the oxidative burst by monocytes from CL patients was higher when compared to monocytes from healthy subjects (HS). Inhibition of the ROS pathway caused a significant decrease in the oxidative burst in L. braziliensis infected monocytes from both groups. In addition, we evaluated the intracellular expression of ROS and NO in L. braziliensis-infected monocytes. Monocytes from CL patients presented high expression of ROS after infection with L. braziliensis. The expression of NO was higher in monocytes from CL patients as compared to expression in monocytes from HS. A strong positive correlation between NO production and lesion size of CL patients was observed. The inhibition of ROS production in leishmania-infected monocytes from CL patients allowed the growth of viable promastigotes in culture supernatants. Thus, we demonstrate that while production of ROS is involved in L. braziliensis killing, NO alone is not sufficient to control infection and may contribute to the tissue damage observed in human CL.

A presença de patógenos respiratórios no biofilme bucal de pacientes com pneumonia nosocomial / Presence of respiratory pathogens in the oral biofilm of patients with nosocomial pneumonia

JUSTIFICATIVA E OBJETIVOS: O paciente internado em unidade de terapia intensiva (UTI), geralmente apresenta higiene bucal insatisfatória, podendo a região orofaríngea ser colonizada por patógenos envolvidos em pneumonia nosocomial. O objetivo deste estudo foi investigar a presença de patógenos respiratórios na cavidade bucal em pacientes em UTI. MÉTODO: Foram incluídos neste estudo transversal, 30 pacientes residentes no município de Nova Friburgo no estado do Rio de Janeiro, com idade entre 18 e 82 anos e média ponderada de 53,53 anos, sendo 17 homens e 13 mulheres, internados na UTI geral, excetuando a unidade coronariana, do Hospital Municipal Raul Sertã, Nova Friburgo, com diagnóstico de pneumonia nosocomial (PN). Foi realizada cultura das amostras do aspirado traqueal para identificar os micro-organismos responsáveis pela PN. Em contrapartida amostras microbiológicas da placa dental supragengival, da língua e do tubo do umidificador, foram analisadas para avaliação da presença do agente etiológico da PN. RESULTADOS: As bactérias mais freqüentemente encontradas no aspirado traqueal dos pacientes foram S. pneumoniae 23,3 por cento (7), P. aeruginosa 20 por cento (6), S. aureus 13,3 por cento (4), Kleibsella pneumoniae 13,3 por cento (4), Candida albicans 6,6 por cento (2), Streptococcus a-hemolítico 6,6 por cento (2), Staphylococcus sp. 6,6 por cento (2), Acinetobacter calcoaceticus - baumanii complex (A. calcoaceticus) em 1 paciente (3,3 por cento do total de pacientes), Eschericia coli (E.coli) 3,3 por cento (1), Enterobacter cloacae (E. cloacae) 3,3 por cento (1). Nesses pacientes, 70 por cento destas bactérias foram encontradas no biofilme dental, 63,33 por cento em amostras da língua, 73,33 por cento nas amostras do tubo do respirador artificial e em 43,33 por cento em todos as áreas simultaneamente. Não foram observadas diferenças significativas nas proporções das amostras dos locais de coleta (p > 0,05). CONCLUSÕES: Os resultados...

BACKGROUND AND OBJECTIVES: Hospitalized patients receiving treatment at intensive care units (ICU) usually show poor oral hygiene, and may have the mouth and oropharingeal region colonized by pathogens involved in nosocomial pneumonia. The presence of these pathogens may increase the risk for respiratory diseases. The aim of this study was to investigate the presence of respiratory pathogens in the oral cavity of hospitalized patients at ICU. METHODS: Were included in the study 30 patients from Hospital Raul Sertã, Nova Friburgo, with the diagnostic of nosocomial pneumonia, and tracheal aspirate samples were cultured to identify the causing microorganisms. In addition, microbiological samples from supragingival dental plaque, tongue and respiratory tube were cultured for the presence of a panel of respiratory pathogens. RESULTS: The most frequently found bacteria in the tracheal aspirate were S. Pneumoniae 23.3 percent (7), P. aeruginosa 20 percent (6), S. aureus 13.3 percent (4), K. pneumoniae 13.3 percent (4), C. albicans 6.6 percent (2), a-hemolytic streptococcus 6.6 percent (2), Staphylococcus sp. 6.6 percent (2), A. calcoaceticus 3.3 percent (1), E. coli 3.3 percent (1) and E. cloacae 3.3 percent (1). 70 percent (21) of these microorganisms were found in the dental biofilm, 63.33 percent (19) in tongue samples; 73.33 percent (22) in the respiratory tube; and 43.33 percent (13) in all sampling sites simultaneously. No differences in proportions could be observed between the sampling sites (p > 0.05) CONCLUSIONS: The results of this study show that respiratory pathogens associated with nosocomial pneumonia are present in the oral biofilm of hospitalized patients in ICU, which may serve as a reservoir for these microorganisms.

[Presence of respiratory pathogens in the oral biofilm of patients with nosocomial pneumonia]. / A presença de patógenos respiratórios no biofilme bucal de pacientes com pneumonia nosocomial.

BACKGROUND AND OBJECTIVES: Hospitalized patients receiving treatment at intensive care units (ICU) usually show poor oral hygiene, and may have the mouth and oropharingeal region colonized by pathogens involved in nosocomial pneumonia. The presence of these pathogens may increase the risk for respiratory diseases. The aim of this study was to investigate the presence of respiratory pathogens in the oral cavity of hospitalized patients at ICU. METHODS: Were included in the study 30 patients from Hospital Raul Sertã, Nova Friburgo, with the diagnostic of nosocomial pneumonia, and tracheal aspirate samples were cultured to identify the causing microorganisms. In addition, microbiological samples from supragingival dental plaque, tongue and respiratory tube were cultured for the presence of a panel of respiratory pathogens. RESULTS: The most frequently found bacteria in the tracheal aspirate were S. Pneumoniae 23.3% (7), P. aeruginosa 20% (6), S. aureus 13.3% (4), K. pneumoniae 13.3% (4), C. albicans 6.6% (2), a-hemolytic streptococcus 6.6% (2), Staphylococcus sp. 6.6% (2), A. calcoaceticus 3.3% (1), E. coli 3.3% (1) and E. cloacae 3.3% (1). 70% (21) of these microorganisms were found in the dental biofilm, 63.33% (19) in tongue samples; 73.33% (22) in the respiratory tube; and 43.33% (13) in all sampling sites simultaneously. No differences in proportions could be observed between the sampling sites (p > 0.05) CONCLUSIONS: The results of this study show that respiratory pathogens associated with nosocomial pneumonia are present in the oral biofilm of hospitalized patients in ICU, which may serve as a reservoir for these microorganisms.