Cytotoxicity evaluation of haloperidol, clozapine and a new molecule with antipsychotic potential, PT-31, in NIH-3T3 cells

Abstract Schizophrenia is an illness that affects 26 million people worldwide. However, conventional antipsychotics present side effects and toxicity, highlighting the need for new antipsychotics. We aimed to evaluate the cytotoxicity of haloperidol (HAL), clozapine (CLO), and a new molecule with antipsychotic potential, PT-31, in NIH-3T3 cells. The neutral red uptake assay and the MTT assay were performed to evaluate cell viability and mitochondrial activity, morphological changes were assessed, and intracellular reactive oxygen species (ROS) detection was performed. HAL and CLO (0.1 µM) showed a decrease in cell viability in the neutral red uptake assay and in the MTT assay. In addition, cell detachment, content decrease, rounding and cell death were also observed at 0.1 µM for both antipsychotics. An increase in ROS was observed for HAL (0.001, 0.01 and 1 µM) and CLO (0.01 and 1 µM). PT-31 did not alter cell viability in any of the assays, although it increased ROS at 0.01 and 1 µM. HAL and CLO present cytotoxicity at 0.1 µM, possibly through apoptosis and necrosis. In contrast, PT-31 does not present cytotoxicity to NIH-3T3 cells. Further studies must be performed for a better understanding of these mechanisms and the potential risk of conventional antipsychotics

Ability of two new thiazolidinediones to downregulate proinflammatory cytokines in peripheral blood mononuclear cells from children with asthma

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

Synthesis and biological evaluation of novel imidazolidine derivatives as candidates to schistosomicidal agents

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.

Cytokine measurements in Brazilian postmenopausal osteoporosis patients reveal high levels of IL-8 / Dosagem de citocinas em pacientes brasileiras com osteoporose pós-menopausa revela altos níveis de IL-8

AIMS: Osteoporosis is a common disease that affects mostly women and has been associated with the immune system. The aims of this study were to evaluate the serum levels of inflammatory cytokines in women with postmenopausal osteoporosis and to investigate their relationship with clinical and laboratory parameters. METHODS: This study recruited patients with postmenopausal osteoporosis (osteoporosis group) and non-osteoporotic postmenopausal women (control group) matched for age. All patients and controls had their bone mineral density measured for the diagnosis of osteoporosis and answered a clinical questionnaire. Blood samples were collected for cytokine measurements. Cytokines IFN-γ, IL-1ß, IL-6, IL-8, IL-9, IL-10, IL-17A, IL-22, IL-27, IL-29, IL-35, and TNF-α were measured by an enzyme-linked immunosorbent assay. RESULTS: Twenty-nine out of the 52 (55.8%) postmenopausal osteoporosis patients showed high levels of IL-8, while no patients from the control group (n=21) showed IL-8 values above the detection limit (p<0.0001). Higher levels of IFN-γ and IL-35 were associated with the control group, with p values of 0.0053 and 0.0214, respectively. In the osteoporosis group, IFN-γ was correlated with longer duration of smoking (p=0.003), IFN-γ and IL-6 were correlated with higher age at menarche (p=0.0454 and p=0.0380), IL-22 was correlated with duration of menopause (p=0.0289) and IL-9 with calcium intake (p=0.019). The other cytokines showed no association or correlation with clinical parameters. CONCLUSIONS: IL-8 was elevated in the serum of patients with postmenopausal osteoporosis, perhaps because it may trigger osteoclast activation and bone wear in osteoporosis. Higher levels of IFN-γ, IL-6, IL-9, IL-22, IL-27, and IL-35 were also associated with the osteoporosis group patients and showed significant correlation with clinical parameters in postmenopausal osteoporosis.

OBJETIVOS: A osteoporose é uma doença comum, que afeta principalmente as mulheres e tem sido associada com o sistema imune. Este estudo objetivou avaliar níveis séricos de citocinas inflamatórias em mulheres pós-menopáusicas com osteoporose, assim como investigar as suas relações com parâmetros clínicos e laboratoriais. MÉTODOS: Este estudo recrutou pacientes com osteoporose pós-menopausa e voluntárias sem a doença, pareadas por idade. Todas as pacientes do grupo com osteoporose e as integrantes do grupo controle passaram pelo exame de mensuração de densidade óssea para diagnosticar a doença e todas responderam a um questionário clínico. Amostras de sangue foram coletadas para as dosagens séricas. As citocinas IFN-γ, IL-1ß, IL-6, IL-8, IL-9, IL-10, IL-17A, IL-22, IL-27, IL-29, IL-35 e TNF-α foram dosadas por ensaio imunoenzimático. RESULTADOS: Vinte e nove entre as 52 (55,8%) pacientes com osteoporose pós-menopausa mostraram altos níveis de IL-8, enquanto nenhuma integrante do grupo controle teve valores de IL-8 acima do nível de detecção do kit (p<0,0001). Altos níveis de IFN-γ and IL-35 foram associados ao grupo controle, com valores de p de 0,0053 and 0,0214 respectivamente. No grupo osteoporose, IFN-γ mostrou correlação com o tempo de duração do tabagismo (p=0,003). IFN-γ e IL-6 foram correlacionadas com a idade de ocorrência da menarca (p=0,0454 e p=0,0380). A citocina IL-22 correlacionou-se com a duração da menopausa (p=0,0289), e a IL-9 com a ingestão de mais cálcio na dieta (p=0,019). As outras citocinas dosadas não mostraram associações ou correlações com os parâmetros clínicos. CONCLUSÕES: A IL-8 mostrou-se elevada no soro das pacientes com osteoporose pós-menopausa, talvez por atuar como um gatilho para a ativação dos osteoclastos e desgaste ósseo que ocorre na osteoporose. Níveis mais altos de IFN-γ, IL-6, IL-9, IL-22, IL-27 e IL-35 também estiveram presentes no soro das pacientes do grupo osteoporose e mostraram associações significativas com os parâmetros clínicos na osteoporose pós-menopausa.

Tegumental Changes in Adult Schistosoma mansoni Induced by a New Imidazolidinic Derivative.

Aims: Verify the potential of the schistosomicidal imidazolidine derivative (5Z)-3-(4- bromo-benzyl)-5-(4-chloro-benzylidene)-4-thioxo-imidazolidin-2-one. Study Design: In this study, we tested the imidazolidinic derivative 3 through in vitro evaluations, cytotoxicity assay and analysis of Scanning Electron Microscopy to verify its therapeutic potential in the treatment of schistosomiasis. Place and Duration of Study: Departamento de Antibióticos, Universidade Federal de Pernambuco (UFPE), Fundação Oswaldo Cruz (FIOCRUZ)/PE and (FIOCRUZ)/BA between January 2013 and march 2014. Methodology: This study was approved by the Ethics Committee on Animal Use Research Center Aggeu Magalhães/Oswaldo Cruz Fundação (CPqAM/FIOCRUZ) authorized by the license No. 21/2011. Male albino Swiss mice were used Mus musculus 25 days old weighing 50 grams. Compound 3 was assayed for its cytotoxicity through cell J774 macrophage lineage. The amount of inhibitory concentration (LC50) was determined by nonlinear regression using the Graph Pad Prism version 5.01. Then the compound was evaluated against adult worms of S. mansoni by performing the activity in vitro at doses 100-20μg/mL and ultrastructural investigation by Scanning Electron Microscopy (SEM) at doses of 100 and 60μg/ml. The PZQ was the positive control of the experiment. Results: The derivative 3 showed LC50 of 29.7±3.9mM. Compound 3 was able to have decreased motility of S. mansoni culminating with a mortality rate of 100% at doses of 60 and 100μg/mL on the fourth day of observation of the experiment. In the SEM, the compound caused various soft tissue changes of S. mansoni parasites such as blistering, destruction of the integument with loss of spines and tubercles, body contraction and windy. Conclusion: The derivative imidazolidine 3 showed a promising schistosomicidal activity in vitro. However, conducting further studies with the completion of work in front of the live schistosomiasis is required.

Activity Anti-Inflammatory and in silico Study of New Thiazolidinedione Derivatives.

Aims: Evaluation of the anti-inflammatory properties of new thiazolidine-2,4-diones derivatives. Study Design: Study the effects of new thiazolidine-2,4-diones derivatives on the inflammatory process. Place and Duration of Study: Departamento de Antibióticos, Universidade Federal de Pernambuco (UFPE), between June 2011 and July 2012. Methodology: Compounds thiazolidine-2,4-diones were tested for anti-inflammatory activity by air pouch model. Swiss albino mice were used for the study. Air cavities were produced by subcutaneous injection of 2.5 mL of sterile air into the intrascapular area of the back. An additional 2.5 mL of air was injected into the cavity every 3 days to keep the space open. Seven days after the initial air injection, 1 mL of a 1% solution of carrageenan dissolved in saline was injected directly into the pouch to produce an inflammatory response. The compoundsthiazolidine-2,4-diones and standard piroxicam were tested at doses of 3 mg/kg body weight. The total number of polymorphonuclear leukocytes (PMNL) was countedusing an improved. Results: The results support the use of these derivatives in inflammatory process. Among the compounds tested the ones that showed a greater effect in inhibiting the migration of neutrophils were the 3a, 3b, 3c, 3d and 3e. The anti-inflammatory effects showed by 3a-j were promising, probably due to the duality of action on PPAR alpha and gamma. Conclusion: In conclusion, this study has shown that the thiazolidine derivatives do possess significant anti-inflammatory effects in laboratory animals. The exact mechanism and the bioactive principles responsible for these actions remain to be explained.

Osteoarticular tuberculosis in an HIV-positive patient: a case report

The authors report a case of a 38-year-old HIV-positive woman, with subcutaneous nodules on the thoracic region with 3 months of evolution. Clinical, laboratory, and epidemiological features were evaluated and associated with apparent damage to the T11-T12 vertebrae, identification by imaging tests, positivity in a polymerase chain reaction-based test, and reactivity to the Mantoux tuberculin skin test (PPD-RT 23). The patient was diagnosed with osteoarticular tuberculosis and received treatment for a year, and clinical cure was achieved.

Anti-inflammatory, Antiarthritic and Antinociceptive Activities of 3,5-Disubstituted Thiazolidine Derivatives.

Aims: The aim of this study was to investigate the anti-inflammatory, antiarthritic and antinociceptive effects of two thiazolidinedione derivatives: 3-(2-bromo-benzyl)-5-(4- methylsufonyl-benzylidene)-thiazolidine-2,4-dione (LPSF/GQ-125) and 3-(2,6-difluorobenzyl)- 5-(4-methylsufonyl-benzylidene)-thiazolidine-2,4-dione (LPSF/GQ-192). Study Design: Study the effects of thiazolidinedione derivatives on the inflammatory process. Place and Duration of Study: Department of Antibiotics of the Federal University of Pernambuco, Brazil, between March 2010 and February 2012. Methodology: The carrageenan-induced air pouch in mice was performed and cytokine levels (TNF-α and IL-1β) were determined. Arthritis was induced in female wistar rats by complete Freund's adjuvant (CFA). To determine the antinociceptive activity, acetic acidinduced nociception and hot plate tests in mice were utilized. Results: Treatment with the compounds reduced leukocyte migration and the release of both TNF-α and IL-1β in the air pouch. Both LPSF/GQ-125 and LPSF/GQ-192 reduced the CFA-induced paw edema and the nociception induced by acetic acid; the hot plate test, however, showed no antinociceptive activity when compared to the control group. Conclusion: These results indicate that LPSF/GQ-125 and LPSF/GQ-192 exhibit promising anti-inflammatory, antinociceptive and antiarthritic activities related to their ability to inhibit TNF-α and IL-1β production as well as reduction of leukocyte migration.

Hydroxychloroquine decreases Th17-related cytokines in systemic lupus erythematosus and rheumatoid arthritis patients

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. .

Relacao estrutura-atividade e mecanismo de acao de anticonvulsivantes derivados do 2-sulfamoilbenzoato. 1. Atividade anti-estricnina / Structure-activity relationship and mechanism of anticonvulsant action of 2-sulfamoylbenzoate derivatives. I. Anti-strychinine activity

Com o objetivo de estudar as relacoes quantitativas entre a estrutura quimica e a atividade anti-estricnina em uma serie de derivados 2-sulfamoilbenzoatos, foram determinados parametros fisico-quimicos obtidos pelo metodo CNDO/2, alguns dos quais, juntamente com a conectividade molecular, permitiram estabelecer os fatores que influem na atividade farmacologica e o possivel mecanismo de acao ao nivel molecular