Abstract Background Oncostatin M (OSM) is a pleiotropic cytokine which, after arterial injury, has proven to be to be rapidly expressed. Objectives To correlate the serum levels of OSM, soluble OSM receptor (sOSMR), and soluble fraction of glycoprotein 130 (sgp130) in patients with coronary artery disease (CAD) with clinical parameters. Methods Levels of sOSMR and sgp130 were evaluated by ELISA and OSM by Western Blot, in patients with CCS (n=100), patients with ACS (n=70), and 64 control volunteers without clinical manifestations of the disease. P-values < 0.05 were considered to be statistically significant. Results CAD patients exhibited significantly lower levels of sOSMR and sgp130 and higher levels of OSM when compared to the controls (both p < 0.0001). Clinical analysis displayed, lower levels of sOSMR in men ([OR] = 2.05, p = 0.026), youth (OR = 1.68, p = 0.0272), hypertensives (OR = 2.19, p = 0.041), smokers (OR = 2.19, p = 0.017), patients that did not present dyslipidemia (OR = 2.32, p = 0.013), patients with Acute Myocardial Infarction [AMI] (OR = 3.01, p = 0.001) and patients not treated with statin (OR = 1.95, p = 0.031), antiplatelet agent (OR = 2.46, p = 0.005), inhibitors of calcium channels (OR = 3.15, p = 0.028), and antidiabetic drugs (OR = 2.97, p = 0.005). The levels of sOSMR were also correlated with gender, age, hypertension, and use of medications in multivariate analysis. Conclusions Our data suggest that the enhanced serum levels of OSM, and decreased levels of sOSMR and sGP130 in patients with cardiac injury may play an important role in the pathophysiological mechanism of the disease. Furthermore, lower levels of sOSMR were associated with gender, age, hypertension, and the use of medications.

Resumo Fundamento A oncostatina M (OSM) é uma citocina pleiotrópica que, após lesão arterial, demonstra ser expressa rapidamente. Objetivos Correlacionar os níveis séricos da OSM, do receptor solúvel de oncostatina M (sOSMR) e da fração solúvel de glicoproteína 130 (sgp130) em pacientes com doença arterial coronariana (DAC) a parâmetros clínicos. Métodos Os níveis de sOSMR e sgp130 foram avaliados por ELISA, enquanto os de OSM foram avaliados por Western Blot, em pacientes com SCC (n=100), pacientes com SCA (n=70) e 64 voluntários do grupo de controle sem manifestações clínicas da doença. Valores de p <0,05 foram considerados estatisticamente significativos. Resultados Pacientes com DAC exibiram níveis significativamente mais baixos de sOSMR e sgp130 e níveis mais altos de OSM em comparação ao grupo de controle (ambos p <0,0001). A análise clínica mostrou níveis mais baixos de sOSMR em homens ([OR] = 2,05, p = 0,026), jovens (OR = 1,68, p = 0,0272), hipertensos (OR = 2,19, p = 0,041), fumantes (OR = 2,19, p = 0,017), pacientes que não apresentavam dislipidemia (OR = 2,32, p = 0,013), pacientes com infarto agudo do miocárdio [IAM] (OR = 3,01, p = 0,001) e pacientes não tratados com estatina (OR = 1,95, p = 0,031), antiplaquetário (OR = 2,46, p = 0,005), inibidores dos canais de cálcio (OR = 3,15, p = 0,028) e antidiabéticos (OR = 2,97, p = 0,005). Os níveis de sOSMR também foram correlacionados a sexo, idade, hipertensão e uso de medicamentos na análise multivariada. Conclusões Nossos dados sugerem que o aumento dos níveis séricos de OSM e a diminuição dos níveis de sOSMR e sGP130 em pacientes com injúria cardíaca podem desempenhar um papel importante no mecanismo fisiopatológico da doença. Além disso, níveis mais baixos de sOSMR foram associados a sexo, idade, hipertensão e uso de medicamentos.

Abstract Oxazolidine derivatives (OxD) have been described as third-line antibiotics and antitumoral agents. The inclusion complexes based on cyclodextrin could improve the solubility and bioavailability of these compounds. A novel synthetic OxD was used, and its inclusion complexes were based on 2-hydroxy-beta-cyclodextrin (2-HPβCD). We conducted an in silico study to evaluate the interaction capacity between OxD and 2-HPβCD. Characterization studies were performed through scanning electron microscopy (SEM), Fourier-transformed infrared (FTIR), nuclear magnetic resonance spectroscopy (1H-NMR), X-ray diffraction (XRD), and thermal analyses. A kinetic study of the OxD was performed, including a cytotoxicity assay using peripheral blood mononuclear cells (PBMCs). The maximum increment of solubility was obtained at 70 mM OxD using 400 mM 2-HPβCD. SEM analyses and FTIR spectra indicated the formation of inclusion complexes. 1H-NMR presented chemical shifts that indicated 1:1 stoichiometry. Different thermal behaviors were obtained. The pharmacokinetic profile showed a short release time. Pure OxD and its inclusion complex did not exhibit cytotoxicity in PBMCs. In silico studies provided a foremost insight into the interactions between OxD and 2-HPβCD, including a higher solubility in water and an average releasing profile without toxicity in normal cells.

Abstract Osteoarthritis (OA) encompasses degeneration of articular cartilage, subchondral bone erosions and sclerosis. Chondrocyte apoptosis and an oxygen-deprived microenvironment are essential factors in OA pathogenesis. PAR-4 (Prostate apoptosis response-4) is a pro-apoptotic protein implicated in many pathologies as well as in chondrocyte cell death mechanism. Vitamin D supplementation has been identified as a therapeutic tool for a variety of inflammatory pathologies. In the present manuscript, we investigated whether first, PAR-4 expression is influenced by chondrocytes in a model of OA, in vitro, and second, whether vitamin D modulates PAR-4 expression in the same model. To test our hypothesis, we used the primary culture of murine chondrocytes isolated from the femoral and tibial condyles of wistar rats. The expression of the pro-inflammatory effect interleukin IL-1β was evaluated in the presence and absence of vitamin D. Western blot and immunofluorescence analysis confirmed protein expression. In the normoxia condition, the chondrocytes expressed PAR-4 in the cell nucleus, and in the hypoxic condition, PAR-4 was expressed in the cell cytoplasm. We disclosed that the treatment with Vitamin D decreased PAR-4 (p= 0.0137) and caspase-3 (p= 0.0007) expression. Thus, the results suggested that PAR-4 and caspase-3 proteins could be potential targets for OA.However, we believe that research is needed to identify the mechanisms implicated in the regulation of PAR-4 in OA.

Abstract INTRODUCTION Herein, the authors describe a simple enhancement to a commercial rapid DNA extraction kit based on simple viral lysis for detecting COVID-19 via RT-qPCR. METHODS After testing several different modifications, the adapted protocol with the best results in preliminary experiments was statistically evaluated in comparison with an automated robotic protocol. RESULTS Processing and testing of 119 nasopharyngeal samples ultimately yielded near-perfect agreement with the automated protocol (κ = 0.981 [95% confidence interval 0.943-1.000]). CONCLUSIONS The low cost and rapidity of the enhanced protocol makes it suitable for adoption in laboratories diagnosing COVID-19, especially those with high demand for examinations.

Allergic asthma is a chronic, complex inflammatory disease of the airway. Despite extensive studies on the immunomodulation of T helper (Th) cell pathways (i.e., Th1 and Th2) in asthma, little is known about the effects of Th17 pathway modulation, particularly that involving peroxisome proliferator-activated receptors (PPARs). In response, two new thiazolidinedione derivatives-namely, LPSF-GQ-147 and LPSF-CR-35 were synthesized and evaluated for their immunomodulatory effects on Th17-related cytokines, including interferon γ (IFNγ), interleukin IL-6, IL-17, and IL-22 in the peripheral blood mononuclear cells of asthmatic children. Both compounds were nontoxic even at high concentrations (i.e., 100 µM). The LPSF-CR-35 compound significantly reduced the levels of IL-17A (p = .039) and IFNγ (p = .032) at 10 µM. For IL-22 and IL-6, significant reduction occurred at 100 µM (p = .039 and p = .02, respectively). Conversely, LPSF-GQ-147 did not significantly inhibit the production of the tested cytokines, the levels of all of which were more efficiently reduced by LPSF-CR-35 than methylprednisolone, the standard compound. Real-time polymerase chain reaction assay confirmed that LPSF-GQ-147 has significant PPARγ modulatory activity. Such data indicate that both LPSF-CR-35 and LPSF-GQ-147 are promising candidates as drugs for treating inflammation and asthma.

ABSTRACT Introduction: Schistosomiasis is an infectious parasitic disease caused by trematodes of the genus Schistosoma, which threatens at least 258 million people worldwide and its control is dependent on a single drug, praziquantel. The aim of this study was to evaluate the anti-Schistosoma mansoni activity in vitro of novel imidazolidine derivatives. Material and methods: We synthesized two novel imidazolidine derivatives: (LPSF/PTS10) (Z)-1-(2-chloro-6-fluorobenzyl)-4-(4-dimethylaminobenzylidene)-5-thioxoimidazolidin-2-one and (LPSF/PTS23) (Z)-1-(2-chloro-6-fluoro-benzyl)-5-thioxo-4-(2,4,6-trimethoxy-benzylidene)-imidazolidin-2-one. The structures of two compounds were determined by spectroscopic methods. During the biological assays, parameters such as motility, oviposition, mortality and analysis by Scanning Electron Microscopy were performed. Results: LPSF/PTS10 and LPSF/PTS23 were considered to be active in the separation of coupled pairs, mortality and to decrease the motor activity. In addition, LPSF/PTS23 induced ultrastructural alterations in worms, after 24 h of contact, causing extensive erosion over the entire body of the worms. Conclusion: The imidazolidine derivatives containing the trimetoxy and benzylidene halogens showed promising in vitro schistosomicidal activity.

A melhoria dos procedimentos diagnósticos que levam a uma identificação prematura do câncer de mama tem elevado a incidência de casos, mas também vêm reduzindo sofrimentos e óbitos em decorrência da doença. Dados da Organização Mundial de Saúde mostram que todo ano, são quase 13 milhões de casos novos de câncer no mundo e 7,5 milhões de mortes por causa da doença. Nas mulheres o câncer mais prevalente é o de mama, no Brasil as taxas de mortalidade continuam elevadas, muito provavelmente porque a doença é diagnosticada em estágios avançados. Neste estudo é feito um levantamento dos exames mais comuns para o diagnóstico do câncer de mama, baseado nos pontos positivos e negativos de cada método e sua efetividade no diagnóstico precoce. Pretende-se assim, identificar os propulsores para inovar nessa área e os métodos mais novos que chegaram ao mercado. Nas análises dos resultados foram observados: que os métodos de diagnósticos mais utilizados são: mamografia, ultrassonografia e ressonância magnética, o mais usado para o rastreamento e diagnóstico precoce é a mamografia, contudo a mesma apresenta-se defasada e ineficaz na diminuição dos óbitos por câncer de mama. Ademais, o acesso ao exame está longe de ser o ideal e a técnica em si depende de uma serie de fatores, tornando-se um procedimento limitado. Concluímos que existe um campo bastante amplo para inovação no diagnóstico em câncer de mama, buscando sempre por técnicas rápidas, práticas, com valores que se encaixem em todos os níveis econômicos e cheguem ate onde os antigos métodos não conseguiram. Os novos métodos são bastante promissores e mostram-se eficazes, se devidamente distribuídos e direcionados.

Improving diagnostic procedures that lead to premature identification of breast cancer has increased the incidence of cases, but have also reducing suffering and deaths from the disease. World Health Organization data show that each year, are nearly 13 million new cases of cancer worldwide and 7.5 million deaths from the disease. In women, the most prevalent cancer is breast cancer, in Brazil mortality rates remain high, most likely because the disease is diagnosed in advanced stages. In this study data is made a survey of the most common tests for the diagnosis of breast cancer, based on the strengths and weaknesses of each method and its effectiveness in the early diagnosis. This is intended to identify the drivers for innovation in this area and the newest methods that hechead the market. in the analyzes of results were observed: the most used diagnostic methods are: mammography, ultrasound and magnetic resonance imaging, the most widely used for screening and early diagnosis is mammography. However it presents itself outdated and ineffective in decreasing deaths breast cancer. in addition, access to the exam is far from ideal and the technique itself depends on a number of factors, making it a flawed procedure. We conclude that there is a very broad field for innovation in the diagnosis of breast cancer, always looking for quick techniques, practices, values that fit all economic levels and to come up to where the old methods have failed. The new methods are very promising and have proven effective if properly distributed and targeted.

OBJECTIVES: Hydroxychloroquine is an antimalarial agent that has been used in systemic lupus erythematosus and rheumatoid arthritis treatment for many years. Recently, novel mechanisms of action have been proposed, thereby broadening the therapeutic perspective of this medication. The purpose of this study was to evaluate the immunomodulatory activity of hydroxychloroquine in T helper 17 (Th17) cytokines in healthy individuals and patients. METHODS: Eighteen female patients with systemic lupus erythematosus (mean age 39.0±12.9 years) and 13 female patients with rheumatoid arthritis (mean age 51.5±7.7 years) were recruited from Universidade Federal de Pernambuco-Brazil. The patients were included after fulfilling four classification criteria for systemic lupus erythematosus or rheumatoid arthritis from the American College of Rheumatology. After being stimulated with phorbol 12-myristate 13-acetate and ionomycin in the absence or presence of different concentrations of hydroxychloroquine, the interleukin 6, 17 and 22 levels were quantified with an enzyme-linked immunosorbent assay in culture supernatants of peripheral blood mononuclear cells from healthy individuals and patients. RESULTS: We demonstrated that in peripheral blood mononuclear cells from healthy volunteers and in systemic lupus erythematosus and rheumatoid arthritis patients, there was a significant reduction in the IL-6, IL-17 and IL-22 supernatant levels after adding hydroxychloroquine. CONCLUSIONS Our in vitro results demonstrated that hydroxychloroquine inhibits IL-6, IL-17 and IL-22 production and contributes to a better understanding of the mechanism of action of this medication.

OBJETIVO: Investigar os níveis de IL-10 e IL-13 no soro de portadores da esquistossomose mansônica na forma hepatoesplênica (EHE), avaliando o papel destas citocinas no desenvolvimento da fibrose hepática. MÉTODOS: O estudo foi prospectivo e analítico, desenvolvido no Departamento de Cirurgia da Universidade Federal de Pernambuco, Laboratório de Imunologia Keizo Asami. Foram estudados três grupos: Grupo I - 25 portadores de esquistossomose mansônica na forma hepatoesplênica e não submetidos a tratamento cirúrgico; Grupo II - 30 submetidos à esplenectomia e ligadura da veia gástrica esquerda; Grupo III - 33 indivíduos sem esquistossomose mansônica na forma hepatoesplênica ou qualquer outra doença ou agravo que comprometesse a reserva funcional hepática. As concentrações séricas de IL-10 e IL-13 foram obtidas pelo método ELISA. Considerando-se a natureza não paramétrica, todas as concentrações foram analisadas pelo teste de Kruskal-Wallis. p<0,05 foi usado para rejeição da hipótese de nulidade. RESULTADOS: As médias das concentrações de IL-10, em ng/mL, no soro foram: GI 50,0 ± 59,0; GII 38,0 ± 270; GIII 38,0 ± 20,0. As concentrações de IL-13, em ng/mL, no soro dos pacientes foram respectivamente: GI 41,0 ± 93,0; GII 16,0 ± 17,0; GIII 18,0 ± 34,0. Não se observou diferença significante entre as médias das concentrações de IL-10 e IL-13 entre os grupos de estudo (p>0,05). CONCLUSÃO: As médias das concentrações séricas de IL-10 e IL-13 foram similares nos três grupos estudados, indicando que, possivelmente, estas citocinas no soro não estejam associadas aos diferentes graus de fibrose de Symmers nos pacientes.

OBJECTIVE: To investigate the serum levels of IL-10 and IL-13 in patients with hepatosplenic schistosomiasis mansoni (HSM), evaluating the role of these cytokines in the development of hepatic fibrosis. METHODS: The study was prospective and analytical, developed at the Department of Surgery, Federal University of Pernambuco, Keizo Asami Laboratory of Immunology. We studied three groups: Group I - 25 patients with hepatosplenic schistosomiasis mansoni who were not submitted to surgery; Group II - 30 individuals who underwent splenectomy and ligature of left gastric vein; Group III - 33 subjects without hepatosplenic schistosomiasis mansoni or any other disease or condition that could compromise the hepatic functional reserve. Serum concentrations of IL-10 and IL-13 were obtained through ELISA. Considering their non-parametric nature, all concentrations were analyzed by Kruskal-Wallis test, with p<0.05 used to reject the null hypothesis. RESULTS: The mean concentrations of IL-10 in ng/mL in serum were GI: 50.0 ± 59.0; GII: 38.0 ± 270; GIII: 38.0 ± 20.0. Concentrations of IL-13 in ng/mL in the serum of patients were respectively: 41.0 ± 93.0 in GI, 16.0 ± 17.0 in GII and 18.0 ± 34.0 in GIII. There was no significant difference between the mean concentrations of IL-10 and IL-13 between the study groups (p> 0.05). CONCLUSION: The mean serum concentrations of IL-10 and IL-13 were similar in all three groups, indicating that possibly the presence of these cytokines in serum is not associated with different degrees of Symmers fibrosis in patients with hepatosplenic schistosomiasis mansoni.