Dynamics of the Trypanosoma cruzi infection in adipose tissue: Assessing gene expression of PNPLA2, FASN, and ACAT1 under Benzonidazole treatment and indirect mononuclear immune cells interaction.

Trypanosoma cruzi is a parasite with a high capacity to adapt to the host. Animal models have already demonstrated that the tropism of this parasite occurs not only in cardiac/digestive tissues but also in adipose tissue (AT). That said, the consequences ofT. cruziinfection for AT and the implications of treatment with Benzonidazole in this tissue are under discussion. Here, we tested the hypothesis that T. cruzi infection in adipose tissue upon treatment with Benzonidazole (Bz) and the interaction of mononuclear immune cells (PBMC) influences the relative expression of ACAT1, FASN, and PNPLA2 genes. Thus, stem cells derived from adipose tissue (ADSC) after adipogenic differentiation were indirectly cultivated with PBMC after infection with the T. cruzi Y strain and treatment with Bz. We use the TcSAT-IAM system and RT-qPCR to evaluate the parasite load and the relative quantification (ΔCt) of the ACAT1, FASN, and PNPLA2 genes. Our results demonstrate that treatment with Bz did not reduce adipocyte infection in the presence (p-value: 0.5796) or absence (p-value: 0.1854) of cultivation with PBMC. In addition, even though there is no statistical difference when compared to the control group (AT), T. cruzi induces the FASN expression (Rq: 14.00). However, treatment with Bz in AT suggests the increases of PNPLA2 expression levels (Rq: 12.58), even in the absence of T. cruzi infection. During indirect cultivation with PBMC, T. cruzi smooths the expression of PNPLA2 (Rq: 0.824) and instigates the expression of ACAT1 (Rq: 1.632) and FASN (Rq: 1.394). Furthermore, the treatment with Bz during infection induces PNPLA2 expression (Rq: 1.871), maintaining FASN expression levels (Rq: 1.334). Given this, our results indicate that treatment with Benzonidazole did not decrease T. cruzi infection in adipose tissue. However, treating the adipocyte cells with Bz during the interaction with PBMC cells influences the lipid pathways scenario, inducing lipolytic metabolism through the expression of PNPLA2.

Induction of SK-MEL-28 Invasion by Brain Cortical Cell-Conditioned Medium Through CXCL10 Signaling.

Melanoma, an infrequent yet significant variant of skin cancer, emerges as a primary cause of brain metastasis among various malignancies. Despite recognizing the involvement of inflammatory molecules, particularly chemokines, in shaping the metastatic microenvironment, the intricate cellular signaling mechanisms underlying cerebral metastasis remain elusive. In our pursuit to unravel the role of cytokines in melanoma metastasis, we devised a protocol utilizing mixed cerebral cortical cells and SK-MEL-28 melanoma cell lines. Contrary to expectations, we observed no discernible morphological change in melanoma cells exposed to a cerebral conditioned medium (CM). However, a substantial increase in both migration and proliferation was quantitatively noted. Profiling the chemokine secretion by melanoma in response to the cerebral CM unveiled the pivotal role of interferon gamma-induced protein 10 (CXCL10), inhibiting the secretion of interleukin 8 (CXCL8). Furthermore, through a transwell assay, we demonstrated that knockdown CXCL10 led to a significant decrease in the migration of the SK-MEL-28 cell line. In conclusion, our findings suggest that a cerebral CM induces melanoma cell migration, while modulating the secretion of CXCL10 and CXCL8 in the context of brain metastases. These insights advance our understanding of the underlying mechanisms in melanoma cerebral metastasis, paving the way for further exploration and targeted therapeutic interventions.

Firework-related ocular trauma in Pernambuco, Brazil / Trauma ocular por fogos de artifício em Pernambuco, Brasil

ABSTRACT Purpose: This study aimed to describe the demographic and clinical characteristics of victims of fireworkrelated ocular trauma treated at the ophthalmologic emergency de partments of two reference centers in Pernambuco, Brazil, and to identify risk factors related to poor visual prognosis. Methods: We retrospectively evaluated the medical records of patients admitted in emergency departments with a report of firework-related trauma between January 2012 and December 2018. Data collected included patient's age, sex, place of origin, month and year of the accident, ocular structures affected, characteristics of the injuries, and type of treatment that patients received. For patients who were followed for >30 days, the final visual acuity and patient's origin were analyzed. Results: Three hundred and seventy eyes from 314 patients were included, of which 248 (79.0%) were male and 160 (51.0%) were from the metropolitan region of Recife. The mean patient age was 25.6 ± 18.8 years. In 56 (17.8%) patients, the ocular trauma was bilateral. A total of 152 (48.4%) cases occurred in June. The most affected sites were the eyelids in 91 (24.6%) eyes and ocular surface in 252 (68.1%). Surgical treatment was required in 87 (23.5%) eyes. After clinical and surgical management, 37 (10.0%) eyes presented final visual acuity of <20/400. Of these, 34 (91.9%) eyes were from patients from the countryside or from another state. Patients from the countryside presented higher risk of developing blindness after a firework trauma than those from the metropolitan area (odds ratio of 5.46). Conclusions: Victims of firework-related ocular trauma were mostly male, from the metropolitan region of Pernambuco state and mainly pediatric patients or economically active. Those coming from the countryside and other states had higher risk of developing blindness

RESUMO Objetivos: Descrever as características demográficas e clínicas das vítimas de trauma ocular por fogos de artifício atendidas nas emergências oftalmológicas de dois centros de referência em Pernambuco e identificar fatores relacionados a mau prognóstico visual. Métodos: Avaliação retrospectiva dos prontuários de pacientes admitidos na emergência oftalmológica com história de trauma por fogos de artifício entre janeiro de 2012 e dezembro de 2018. A coleta de dados incluiu idade, gênero, procedência, mês e ano do acidente, estruturas oculares acometidas e características das lesões, além do tipo de tratamento a que os pacientes foram submetidos. Naqueles pacientes acompanhados por mais de 30 dias, analisou-se a acuidade visual final e a associação com sua procedência. Resultados: Foram incluídos 370 olhos de 314 pacientes. Destes, 248 (79,0%) vítimas eram do sexo masculino e 160 (51,0%) da região metropolitana do Recife, com uma média de idade de 25.6 ± 18.8 anos. Em 56 (17,8%) dos casos o trauma foi bilateral. No mês de junho ocorreu um total de 152 (48,4%) casos. Os sítios mais acometidos foram pálpebras em 91 (24,6%) olhos e superfície ocular em 252 (68,1%). O tratamento cirúrgico foi necessário em 87 (23,5%) olhos. Após manejo clínico-cirúrgico, 37 (10.0%) olhos desenvolveram visão pior do que 20/400. Destes, 34 (91,9%) olhos eram de pacientes do interior do estado de Pernambuco ou de outro estado. Os pacientes provenientes do interior do estado apresentaram maior chance de desenvolver cegueira quando comparados aos que eram provenientes da região metropolitana (Odds Ratio de 5,46). Conclusões: As vítimas de trauma ocular por fogos de artificio foram em sua maioria do sexo masculino, procedentes da região metropolitana do estado e das faixas etárias pediátrica e economicamente ativa. Aqueles provenientes do interior ou de outros estados apresentaram maior chance de desenvolver cegueira.

Firework-related ocular trauma in Pernambuco, Brazil.

PURPOSE: This study aimed to describe the demographic and clinical characteristics of victims of fireworkrelated ocular trauma treated at the ophthalmologic emergency de partments of two reference centers in Pernambuco, Brazil, and to identify risk factors related to poor visual prognosis. METHODS: We retrospectively evaluated the medical records of patients admitted in emergency departments with a report of firework-related trauma between January 2012 and December 2018. Data collected included patient's age, sex, place of origin, month and year of the accident, ocular structures affected, characteristics of the injuries, and type of treatment that patients received. For patients who were followed for >30 days, the final visual acuity and patient's origin were analyzed. RESULTS: Three hundred and seventy eyes from 314 patients were included, of which 248 (79.0%) were male and 160 (51.0%) were from the metropolitan region of Recife. The mean patient age was 25.6 ± 18.8 years. In 56 (17.8%) patients, the ocular trauma was bilateral. A total of 152 (48.4%) cases occurred in June. The most affected sites were the eyelids in 91 (24.6%) eyes and ocular surface in 252 (68.1%). Surgical treatment was required in 87 (23.5%) eyes. After clinical and surgical management, 37 (10.0%) eyes presented final visual acuity of <20/400. Of these, 34 (91.9%) eyes were from patients from the countryside or from another state. Patients from the countryside presented higher risk of developing blindness after a firework trauma than those from the metropolitan area (odds ratio of 5.46). CONCLUSIONS: Victims of firework-related ocular trauma were mostly male, from the metropolitan region of Pernambuco state and mainly pediatric patients or economically active. Those coming from the countryside and other states had higher risk of developing blindness.

Benznidazole treatment decreases IL-6 levels in Trypanosoma cruzi-infected human adipocytes differentiated from adipose tissue-derived stem cells.

BACKGROUND: Trypanosoma cruzi, which causes Chagas disease (CD), is a versatile haemoparasite that uses several strategies to evade the host's immune response, including adipose tissue (AT), used as a reservoir of infection. As it is an effective barrier to parasite evasion, the effectiveness of the drug recommended for treating CD, Benznidazole (BZ), may be questionable. OBJECTIVE: To this end, we evaluated the parasite load and immunomodulation caused by BZ treatment in the culture of adipocytes differentiated from human adipose tissue-derived stem cells (ADSC) infected with T. cruzi. METHODS: The ADSC were subjected to adipogenic differentiation. We then carried out four cultures in which we infected the differentiated AT with trypomastigote forms of the Y strain of T. cruzi and treated them with BZ. After the incubation, the infected AT was subjected to quantitative polymerase chain reaction (qPCR) to quantify the parasite load and transmission electron microscopy (TEM) to verify the infection. The supernatant was collected to measure cytokines, chemokines, and adipokines. FINDINGS: We found elevated secretion of IL-6, CXCL-10/IP-10, CCL2/MCP-1, CCL5/RANTES, and leptin in infected fat cells. However, treatment with BZ promoted a decrease in IL-6. MAIN CONCLUSION: Therefore, we believe that BZ has a beneficial role as it reduces inflammation in infected fat cells.

Benznidazole treatment decreases IL-6 levels in Trypanosoma cruzi-infected human adipocytes differentiated from adipose tissue-derived stem cells

BACKGROUND Trypanosoma cruzi, which causes Chagas disease (CD), is a versatile haemoparasite that uses several strategies to evade the host's immune response, including adipose tissue (AT), used as a reservoir of infection. As it is an effective barrier to parasite evasion, the effectiveness of the drug recommended for treating CD, Benznidazole (BZ), may be questionable. OBJECTIVE To this end, we evaluated the parasite load and immunomodulation caused by BZ treatment in the culture of adipocytes differentiated from human adipose tissue-derived stem cells (ADSC) infected with T. cruzi. METHODS The ADSC were subjected to adipogenic differentiation. We then carried out four cultures in which we infected the differentiated AT with trypomastigote forms of the Y strain of T. cruzi and treated them with BZ. After the incubation, the infected AT was subjected to quantitative polymerase chain reaction (qPCR) to quantify the parasite load and transmission electron microscopy (TEM) to verify the infection. The supernatant was collected to measure cytokines, chemokines, and adipokines. FINDINGS We found elevated secretion of IL-6, CXCL-10/IP-10, CCL2/MCP-1, CCL5/RANTES, and leptin in infected fat cells. However, treatment with BZ promoted a decrease in IL-6. MAIN CONCLUSION Therefore, we believe that BZ has a beneficial role as it reduces inflammation in infected fat cells.

Evaluation of cytotoxic, immunomodulatory and antibacterial activities of aqueous extract from leaves of Conocarpus erectus Linnaeus (Combretaceae).

OBJECTIVES: This work evaluated the antibacterial activity, cytotoxicity and immunomodulatory effect on human peripheral blood mononuclear cells (PBMCs) promoted by aqueous extract from Conocarpus erectus leaves (AELCe). METHODS: The extract was characterized by thin layer chromatography and ultraperformance liquid chromatography coupled to mass spectrometry (UPLC-MS). Cytotoxicity of AELCe (6.25-50 µg/ml) was investigated using annexin V and propidium iodide. Cytokine and nitric oxide levels in PBMCs culture supernatants exposed or not to AELCe (12.5 µg/ml) were determined, and antibacterial activity was evaluated by disc diffusion and broth microdilution methods. KEY FINDINGS: AELCe contained 3',4'-OH flavonoids, phenylpropanoglycosides, saponins, polymeric proanthocyanidins and hydrolysable tannins. Moreover, 10 other compounds were identified through UPLC-MS technique. AELCe did not affect lymphocyte viability at 6.25 and 12.5 µg/ml. IL-2, IL-10, TNF-α, IFN-γ and nitric oxide was produced in higher levels by cells treated with AELCe. Proliferation and activation of CD8+ T lymphocytes were also stimulated. AELCe showed bacteriostatic activity against clinical and antibiotic-resistant strains of Staphylococcus aureus (MIC between 250 and 1000 µg/ml). CONCLUSIONS: AELCe showed a moderate bacteriostatic activity and promoted an immunomodulatory status through higher production of Th1 cytokines, nitric oxide release and T CD8+ lymphocytes stimulation.

Expression of co-stimulatory molecules CD80 and CD86 is altered in CD14 + HLA-DR + monocytes from patients with Chagas disease following induction by Trypanosoma cruzi recombinant antigens.

INTRODUCTION: The relationships between monocytes and lymphocytes through MHC class II molecules and costimulatory, are of utmost importance for the production of an efficient immune response. In this work, we assessed the expression of surface molecules CD80 and CD86 on CD14+HLA-DR+ monocytes from patients with Chagas disease. METHODS:: The study population consisted of 31 patients with chronic clinical forms of Chagas disease. Patient blood samples were cultured in the presence of recombinant cytoplasmic repetitive antigen (CRA) and flagellar repetitive antigen (FRA). RESULTS:: We found considerable differences in the expression profile of surface molecules involved in antigen presentation. CONCLUSIONS:: CRA and FRA may contribute to host immune response evasion by Trypanozoma cruzi.

Avaliação dos monócitos entre as formas clínicas crônicas da doença de Chagas antes e após estímulo in vitro com os antígenos recombinantes CRA e FRA de Trypanosoma cruzi / Assessing of monocytes between the clinical forms of chronic Chagas' disease before and after in vitro stimulation with CRA and FRA recombinant antigens of Trypanosoma cruzi

As interações moleculares entre os monócitos e os linfócitos são de extrema importância para a produção de uma resposta imune eficiente. Nesse contexto nos propomos avaliar a expressão da molécula de superfície HLA-DR+ em monócitos CD14+, e das moléculas co-estimulatórias CD80+ e CD86+ e a produção das citocinas IL-10, IFN-gama, TNF-alfa e IL-12 por monócitos CD14+ HLA-DR+ de portadores de formas clínicas crônicas da doença de Chagas. O grupo de indivíduos acometidos cronicamente pela infecção chagásica foi composto por 10 portadores da forma cardíaca (FC) sem dilatação cardíaca (FC1), 14 portadores da forma cardíaca com dilatação cardíaca (FC2) e 7 portadores da forma indeterminada (FI). O sangue total destes indivíduos foi submetido à cultura na presença dos antígenos CRA ou FRA, específicos ao Trypanosoma cruzi. Observamos que o aumento na expressão da molécula co-estimulatória CD80+ associado à diminuição da expressão de CD86+ no grupo de indivíduos FC, quando comparamos os contextos ex vivo e após estímulo com CRA ou FRA, aponta um possível envolvimento desses antígenos nos mecanismos de fuga do sistema imune do hospedeiro pelo T. cruzi. Correlações positivas entre citocinas inflamatórias e anti-inflamatórias foram observadas entre os portadores da FI e FC1, indicando um controle do parasitismo, concomitantemente a uma modulação da resposta inflamatória. No grupo de pacientes FC2 observamos uma predominância de uma correlação positiva entre as citocinas inflamatórias, bem como de correlação positiva entre citocinas Th1 e Th2, indicando que há predomínio de um perfil do tipo Th1, mas com indícios de mecanismo de regulação da resposta inflamatória exacerbada. Nossos resultados sugerem que os antígenos CRA e FRA, por estarem presentes no parasito, poderiam atuar no processo de apresentação e ativação celular durante a cronicidade da doença, sobretudo nos indivíduos portadores da FC.