Diazepam, in a single dose, inhibits cellular chemotaxis, macrophage stimulation, and TNF-α activity in LPS-induced acute inflammatory responses in mice

Os benzodiazepínicos estão entre as drogas mais freqüentemente prescritas em razão de suas propriedades ansiolíticas. O objetivo deste trabalho foi avaliar a influência do diazepam sobre a resposta inflamatória peritoneal aguda induzida por lipopolissacarídeo. Para tanto, camundongos Swiss foram tratados com diazepam (1 ou 10 mg/kg de peso), em dose única, por via subcutânea, uma hora antes do desafio intraperitoneal com lipopolissacarídeo bacteriano. Após 16 horas do desafio, os animais foram sacrificados, coletando-se os lavados peritoneais para determinação do número total de células e das subpopulações de mononucleares e polimorfonucleares, além da atividade de TNF-α e da porcentagem de macrófagos espraiados. Observou-se que o tratamento com diazepam, nas doses de 1 ou 10 mg/kg, reduziu significativamente a porcentagem de macrófagos estimulados por LPS e a liberação de TNF-α independente de estímulo. Houve também significativa redução da migração de leucócitos nos animais estimulados com LPS e tratados com 10 mg/kg de diazepam em relação aqueles não tratados. Concluímos que a administração do diazepam, em dose única, pode influenciar significativamente o influxo celular, a estimulação de macrófagos e a atividade de TNF-α na resposta inflamatória aguda induzida por LPS em camundongos, com possíveis implicações na eficiência da resposta anti-infecciosa.Benzodiazepines are one of the most frequently prescribed drugs due to their anxiolytic properties. The aim of this study was to evaluate the effects of diazepam on lipopolysaccharide-induced peritoneal acute inflammatory responses. Swiss mice were treated with diazepam in a single dose of 1 or 10 mg/kg- subcutaneously 1 h before an intraperitoneal injection of lipopolysaccharide or sterile saline solution. The mice were killed 16 h after and the cells were washed from the peritoneal cavity to determine the total number of cells and the mononuclear and polimorfonuclear subpopulations, as well as the TNF-alpha activity and percentage of spread macrophages. Our results showed that the diazepam treatment (1 and 10 mg/kg) induced a significant reduction in the LPS-induced macrophage stimulation and TNF-α activity. Diazepam (10 mg/kg) also reduced the inflammatory cellular migration when compared to the control. It can be concluded that the diazepam treatment in a single dose is able to influence the inflammatory cellular influx, macrophage stimulation and TNF-α activity in the acute inflammatory response in mice, having possible implications on the anti-infectious response efficiency

Efeito de tratamentos térmicos após a polimerização sobre a citotoxicidade de duas resinas acrílicas para base de próteses

INTRODUCTION: Most denture base acrylic resins have polymethylmethacrylate in their composition. Several authors have discussed the polymerization process involved in converting monomer into polymer because adequate polymerization is a crucial factor in optimizing the physical properties and biocompatibility of denture base acrylic resins. To ensure the safety of these materials, in vitro cytotoxicity assays have been developed as preliminary screening tests to evaluate material biocompatibility. ³H-thymidine incorporation test, which measures the number of cells synthesizing DNA, is one of the biological assays suggested for cytotoxicity testing. AIM: The purpose of this study was to investigate, using ³H-thymidine incorporation test, the effect of microwave and water-bath post-polymerization heat treatments on the cytotoxicity of two denture base acrylic resins. MATERIALS AND METHODS: Nine disc-shaped specimens (10 x 1 mm) of each denture base resin (Lucitone 550 and QC 20) were prepared according to the manufacturers' recommendations and stored in distilled water at 37ºC for 48 h. The specimens were assigned to 3 groups: 1) post-polymerization in a microwave oven for 3 min at 500 W; 2) post-polymerization in water-bath at 55º C for 60 min; and 3) without post-polymerization. For preparation of eluates, 3 discs were placed into a sterile glass vial with 9 mL of Eagle's medium and incubated at 37ºC for 24 h. The cytotoxic effect of the eluates was evaluated by ³H-thymidine incorporation. RESULTS: The results showed that the components leached from the resins were cytotoxic to L929 cells, except for the specimens heat treated in water bath (pINTRODUÇÃO: A maioria das resinas acrílicas utilizadas para confecção de bases de próteses é composta pelo polimetacilato de metila. Muitos autores têm discutido o processo de polimerização dessas resinas em relação à conversão do monômero em polímero devido a sua importância na melhora da biocompatibilidade e das propriedades físicas. Para assegurar a utilização desses materiais, testes preliminares de citotoxicidade in vitro têm sido desenvolvidos para avaliação da biocompatibilidade. Um dos ensaios biológicos sugeridos para a análise da citotoxicidade é o teste de incorporação de ³H-timidina, o qual mede o número de células por meio da síntese de DNA. OBJETIVO: O objetivo do presente estudo foi avaliar, por meio do teste de incorporação de ³H-timidina, a citotoxicidade de duas resinas acrílicas para base de próteses submetidas aos tratamentos em microondas e em banho de água após a polimerização. MATERIAL E MÉTODO: Nove corpos-de-prova em forma de discos (10 x 1 mm) foram confeccionados com as resinas acrílicas Lucitone 550 e QC 20 de acordo com as instruções dos fabricantes, e foram armazenados em água destilada a 37ºC por 48 h. Os corpos-de-prova foram divididos em três grupos: 1) tratamento em forno de microondas por 3 min a 500 W; 2) tratamento em banho de água a 55ºC por 60 min; e 3) sem tratamento térmico. Extratos foram preparados pela colocação de 3 discos em tubos de ensaio estéreis com 9 mL de meio de cultura Eagle e incubação a 37ºC por 24 h. O efeito citotóxico dos extratos foi avaliado utilizando o teste de incorporação de ³H-timidina. RESULTADOS: Os resultados indicaram que os componentes liberados pelas resinas foram citotóxicos para as células L929 exceto para as amostras tratadas em banho de água (

Interplay among the oral microbiome, oral cavity conditions, the host immune response, diabetes mellitus, and its associated-risk factors : an overview

This comprehensive review of the literature aimed to investigate the interplay between the oral microbiome, oral cavity conditions, and host immune response in Diabetes mellitus (DM). Moreover, this review also aimed to investigate how DM related risk factors, such as advanced age, hyperglycemia, hyperlipidemia, obesity, hypertension and polycystic ovary syndrome (PCOS), act in promoting or modifying specific mechanisms that could potentially perpetuate both altered systemic and oral conditions. We found that poorly controlled glycemic index may exert a negative effect on the immune system of affected individuals, leading to a deficient immune response or to an exacerbation of the inflammatory response exacerbating DM-related complications. Hyperglycemia induces alterations in the oral microbiome since poor glycemic control is associated with increased levels and frequencies of periodontal pathogens in the subgingival biofilm of individuals with DM. A bidirectional relationship between periodontal diseases and DM has been suggested: DM patients may have an exaggerated inflammatory response, poor repair and bone resorption that aggravates periodontal disease whereas the increased levels of systemic pro-inflammatory mediators found in individuals affected with periodontal disease exacerbates insulin resistance. SARSCoV-2 infection may represent an aggravating factor for individuals with DM. Individuals with DM tend to have low salivary flow and a high prevalence of xerostomia, but the association between prevalence/experience of dental caries and DM is still unclear. DM has also been associated to the development of lesions in the oral mucosa, especially potentially malignant ones and those associated with fungal infections. Obesity plays an important role in the induction and progression of DM. Co-affected obese and DM individuals tend to present worse oral health conditions. A decrease in HDL and, an increase in triglycerides bloodstream levels seem to be associated with an increase on the load of periodontopathogens on oral cavity. Moreover, DM may increase the likelihood of halitosis. Prevalence of impaired taste perception and impaired smell recognition tend to be greater in DM patients. An important interplay among oral cavity microbiome, DM, obesity and hypertension has been proposed as the reduction of nitrate into nitrite, in addition to contribute to lowering of blood pressure, reduces oxidative stress and increases insulin secretion, being these effects desirable for the control of obesity and DM. Women with PCOS tend to present a distinct oral microbial composition and an elevated systemic response to selective members of this microbial community, but the association between oral microbiome, PCOS are DM is still unknown. The results of the studies presented in this review suggest the interplay among the oral microbiome, oral cavity conditions, host immune response and DM and some of the DM associated risk factors exist. DM individuals need to be encouraged and motivated for an adequate oral health care. In addition, these results show the importance of adopting multidisciplinary management of DM and of strengthening physicians-dentists relationship focusing on both systemic and on oral cavity conditions of DM patients

A DONOR-DEPENDENT SUBSET OF CYTOKINE-INDUCED KILLER (CIK) CELLS EXPRESS CD16 AND CAN BE RETARGETED TO EXERT A POTENT ANTIBODY-DEPENDENT CELL-MEDIATED CYTOTOXICITY (ADCC)

Cancer adoptive cell therapy (ACT) relies on the infusion of immune cell populations mediating direct antitumor effects, such as cytotoxic CD8+ T lymphocytes (CTL), natural killer (NK) cells and Cytokine-Induced Killer (CIK) cells. In this study, we aimed at improving CIK cell potential for adoptive immunotherapy strategies. CIK cells are a heterogeneous population of ex vivo expanded lymphocytes, which share phenotypic and functional features with both NK and T cells. They exert a potent MHC-independent antitumor activity against both hematological and solid malignancies, but not normal tissues and hematopoietic precursors. Several clinical trials have demonstrated the feasibility and the therapeutic efficacy together with low toxicity of CIK cells infusion, supporting CIK cells as a very promising cell population for adoptive immunotherapy. In this work, CIK cells were obtained from PBMCs of healthy donors by the timed addition of IFN-γ, anti-CD3 antibody and IL-2. Analyzing their phenotype, we demonstrated for the first time a relevant expression of CD16 in a donor-dependent manner and, based on this observation, we proved the ability of CIK cells to kill tumors by an Antibody-Dependent Cell-mediated Cytotoxicity (ADCC) mechanism. Indeed, the concurrent administration of clinically therapeutic mAbs, such as trastuzumab or cetuximab, led to a significant improvement of their antitumor activity in vitro against both ovarian and breast cancer cell lines. To formally prove that the CD16 receptor is functional and directly involved in the ADCC, an anti-CD16 blocking antibody was added to the assays. NK cell depletion from bulk cultures confirmed that the ADCC activity is accountable to the CIK CD16+ subpopulation. This novel function of CIK cells, never exploited before, was assessed for therapeutic efficacy in mouse models of human ovarian carcinoma xenografted in NOD/SCID common γ chain knockout (NSG) mice. Co-administration of CIK cells and mAbs significantly increased the survival of tumor-bearing mice, as compared to animals receiving CIK cells alone. CIK cell antitumor activity in vitro was also enhanced by the combination with bispecific antibodies and immunoligands, which are able to target both a tumor-associated antigen and activating receptors expressed by effector cells. Taken together, these data envisage new perspectives for adoptive immunotherapy where antigen-specific retargeting of T cells can be achieved by a combination therapy with clinical-grade monoclonal antibodies already widely used in cancer therapy, and CIK cell populations that are easily expandable in very large numbers, inexpensive, safe and do not require genetic manipulations. In conclusion, this new therapeutic strategy for the ACT treatment of different types of tumors could find wide implementation and application, and be expanded to the use of additional therapeutic antibodies

Immunological response in mice bearing LM3 breast tumor undergoing Pulchellin treatment

Background: Ribosome-inactivating proteins (RIP) have been studied in the search for toxins that could be used as immunotoxins for cancer treatment. Pulchellin, a type 2 RIP, is suggested to induce immune responses that have a role in controlling cancer. Methods: The percentage of dendritic cells and CD4+ and CD8+ T cells in the spleen (flow cytometry), cytokines’ release by PECs and splenocytes (ELISA) and nitric oxide production by PECs (Griess assay) were determined from tumor-bearing mice injected intratumorally with 0.1 ml of pulchellin at 0.75 μg/kg of body weight. Statistical analysis was performed by one-way ANOVA with Tukey’s post hoc test. Results: Pulchellin-treated mice showed significant immune system activation, characterized by increased release of IFN-γ and Th2 cytokines (IL-4 and IL-10), while IL-6 and TGF-β levels were decreased. There was also an increase in macrophage’s activation, as denoted by the higher percentage of macrophages expressing adhesion and costimulatory molecules (CD54 and CD80, respectively). Conclusions: Our results suggest that pulchellin is promising as an adjuvant in breast cancer treatment.Fil: de Matos, Djamile Cordeiro. Universidade de Sao Paulo; BrasilFil: Abreu de Ribeiro, Livia Carolina. Universidade de Sao Paulo; BrasilFil: Tansini, Aline. Universidade de Sao Paulo; BrasilFil: Ferreira, Lucas Souza. Universidade de Sao Paulo; BrasilFil: Polesi Placeres, Marisa Campos. Universidade de Sao Paulo; BrasilFil: Colombo, Lucas Luis. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Iracilda Zeppone, Carlos. Universidade de Sao Paulo; Brasi

Foxp3 Silencing with Antisense Oligonucleotide Improves Immunogenicity of an Adjuvanted Recombinant Vaccine against Sporothrix schenckii

Background: In recent years, there has been great interest in developing molecular adjuvants based on antisense oligonucleotides (ASOs) targeting immunosuppressor pathways with inhibitory effects on regulatory T cells (Tregs) to improve immunogenicity and vaccine efficacy. We aim to evaluate the immunostimulating effect of 2′OMe phosphorothioated Foxp3-targeted ASO in an antifungal adjuvanted recombinant vaccine. Methods: The uptake kinetics of Foxp3 ASO, its cytotoxicity and its ability to deplete Tregs were evaluated in murine splenocytes in vitro. Groups of mice were vaccinated with recombinant enolase (Eno) of Sporothix schenckii in Montanide Gel 01 adjuvant alone or in combination with either 1 µg or 8 µg of Foxp3 ASO. The titers of antigen-specific antibody in serum samples from vaccinated mice (male C57BL/6) were determined by ELISA (enzyme-linked immunosorbent assay). Cultured splenocytes from each group were activated in vitro with Eno and the levels of IFN-γ and IL-12 were also measured by ELISA. The results showed that the anti-Eno antibody titer was significantly higher upon addition of 8 µM Foxp3 ASO in the vaccine formulation compared to the standard vaccine without ASO. In vitro and in vivo experiments suggest that Foxp3 ASO enhances specific immune responses by means of Treg depletion during vaccination. Conclusion: Foxp3 ASO significantly enhances immune responses against co-delivered adjuvanted recombinant Eno vaccine and it has the potential to improve vaccine immunogenicity

A construção das personagens em The Zoo Story: uma abordagem sistêmico-funcional

Este trabalho busca mostrar como se dá a interação das personagens Peter e Jerry na peça The Zoo Story, de Edward Albee, tomando como aporte teórico a linguística sistêmico-funcional. Foram analisadas e quantificadas as trocas de informação e de bens e serviços e do recurso à modalidade nas falas das personagens, bem como dos tipos de processo associados a cada uma nas rubricas do texto, de modo a observar como se deu a construção dessas personagens e a dinâmica de sua interação ao longo da peça. A contabilização das funções atribuídas ao Finito mostrou que as personagens deram primazia a localizar aquilo que expressavam no tempo e no espaço; no entanto, os casos de modalidade e de modalização e modulação elucidaram um forte contraste na personalidade de ambas, reflexo de suas diferentes condições socioeconômicas. Finalmente, as manifestações das personagens no plano físico, através da quantificação dos tipos de processos associados a elas, serviram para mostrar as atitudes de cada uma com relação a sua contraparte, atitudes essas nem sempre manifestadas verbalmente, daí a utilidade de observar os processos que guiam as rubricas do texto. Este trabalho busca elucidar algumas propriedades das metafunções experiencial e interpessoal da LSF e mostrar como ela pode ser aplicada à análise do texto literário, revelando aspectos não somente formais do texto, mas também a maneira como as personagens são construídas em relação umas às outras.This paper seeks to show how the interaction between the characters Peter and Jerry takes place in the play The Zoo Story, by Edward Albee, taking systemic-functional linguistics as a theoretical contribution. The exchanges of information and goods and services and the use of modality in the characters’ dialogues were analyzed and quantified, as well as the types of process associated with each one in the performance rubrics, in order to observe the construction of these characters and the dynamics of their interaction throughout the play. The accounting of the functions attributed to Finite showed that the characters gave priority to locating what they expressed in time and space; however, modality and modulation cases elucidated a strong contrast in the personality of both, reflecting their different socioeconomic conditions. Finally, the physical manifestations of the, through the quantification of the types of processes associated with them, served to show the attitudes of each one in relation to their counterpart, attitudes not always manifested verbally, hence the utility of observing the processes that guide the text rubrics. This paper seeks to elucidate some properties of the experiential and interpersonal metafunctions and to show how they can be applied to the analysis of the literary text, revealing not only formal aspects of the text but also the way the characters are constructed in relation to one another.info:eu-repo/semantics/publishedVersio

Citotoxicidade de Styrax camporum Pohl em macrófagos peritoneais e a liberação de H2O2

The immunological response includes wide contexts involving several cells, and the macrophage is crucial in the cellular immune response. Several stimuli to macrophage membrane may induce the liberation of H2O2, contributing to antibacterial and cytotoxicical actions. Nowadays, there is a tendency to study natural products to verify their capacity of acting in the immune system. This study evaluated the citotoxicity of the bulk extract and the hexanic and acetic fractions extracted from Styrax camporum Pohl (Styracaceae) and the production of H2O2, on murine peritonal macrophages cultures exposed to fractions extracted from this plant. The results showed that the fraction HX 2 mg/ml produced the liberation of H 2O2 in high concentrations and to 4 mg/ml was observed high citotoxicity. The fractions AC did not produce the liberation of H 2O2 and EB was produced in low levels. We conclude that this HX is a potent stimulator of macrophage

Interação das enterotoxinas estafilocócicas com o sistema imune do hospedeiro

Staphylococcal enterotoxins are among the most common etiologic agents that cause food poisoning and, possibly, nonmenstrual toxic shock syndrome. These enterotoxins are also called superantigens because they are potent T cell and macrophages activators. The superantigens bind directly to the major histocompatibility complex class II molecules on antigen-presenting cells and stimulate T cells expressing specific Vβ elements in the cell receptors. Excessive production of cytokines by these cells and macrophages are responsible for the pathogenesis of food poisoning. These cytokine include tumor necrosis factor (TNF)-α, interferon (IFN)-γ and interleukin (IL)-1, proinflamatory mediators with potent immunoenhancing effects; the nitric oxide (NO). It still has both effects citotoxic and regulatory roles in immune function