ABSTRACT Objective To determine the role of the AKT pathway in the regulating of natural Killer-induced apoptosis of acute myeloid leukemia cells and to characterize the associated molecular mechanisms. Methods BALB/c nude mice were injected with HL60 cells to induce a xenogenic model of subcutaneous leukemic tumors. Mice were treated with perifosine, and their spleens were analyzed using biometry, histopathology, and immunohistochemistry. Gene expression analysis in leukemia cells was performed by real-time PCR. Protein analysis of leukemia and natural Killer cells was performed by flow cytometry. AKT inhibition in HL60 cells, followed by co-culture with natural Killer cells was performed to assess cytotoxicity. Apoptosis rate was quantified using flow cytometry. Results Perifosine treatment caused a reduction in leukemic infiltration in the spleens of BALB/c nude mice. In vitro , AKT inhibition reduced HL60 resistance to natural Killer-induced apoptosis. AKT inhibition suppressed the immune checkpoint proteins PD-L1, galectin-9, and CD122 in HL60 cells, but did not change the expression of their co-receptors PD1, Tim3, and CD96 on the natural Killer cell surface. In addition, the death receptors DR4, TNFR1, and FAS were overexpressed by AKT inhibition, thus increasing the susceptibility of HL60 cells to the extrinsic pathway of apoptosis. Conclusion The AKT pathway is involved in resistance to natural Killer-induced apoptosis in HL60 cells by regulating the expression of immune suppressor receptors. These findings highlight the importance of AKT in contributing to immune evasion mechanisms in acute myeloid leukemia and suggests the potential of AKT inhibition as an adjunct to immunotherapy.

ABSTRACT The Scientists of Tomorrow/ Cientistas do Amanhã project is an immersive science training program developed by the Program of Post-Graduation in Health Sciences at Hospital Israelita Albert Einstein. This program was conducted in partnership with Volunteering and Escola Municipal de Ensino Fundamental Professor Paulo Freire in Paraisópolis, São Paulo, Brazil. The Scientists of Tomorrow Program comprised a short training period conducted in May 2022 involving 37 students, and a long training period from August to December 2022, which included 15 students. It aimed to popularize science through practical activities; transfer knowledge to young students; sensitize and guide them to pursue academic-scientific careers; reduce stereotypes about scientific work and scientists; and help students understand the social, political, and ethical roles of science within society. All activities were led by postgraduate students and professors from our postgraduate program, physicians, nurses, physiotherapists, biomedicals, and veterinarians from Hospital Israelita Albert Einstein, as well as medical students from Faculdade Israelita de Ciências da Saúde Albert Einstein . Activities in the short training included lectures on cinema and science, strategies to combat fake news, non-violent communication, innovation, design-thinking framework, and developing a scientific project. During the long training period, discussions were focused on nanotechnology, animal research, big data, bioinformatics, meditation, blood and bone marrow donation, telemedicine, sex and sexually-transmitted infections, rehabilitation, career opportunities, and scientific integrity. In addition, practical activities were further expanded using optical and confocal microscopy, cytometry, and basic concepts regarding the structure and function of living cells. The program also included the launching of the open-air outreach Education E-natureza activity, which turned students into ambassadors of nature. In conclusion, the Scientists of Tomorrow Program was innovative and enabled young students to learn that science is a collective activity that can enhance public health.

RESUMO Objetivo O objetivo do estudo foi analisar as evidências sobre miRNAs como biomarcadores para o diagnóstico de endometriose, bem como levantar informações sobre os desafios e as estratégias necessárias para tornar essas moléculas ferramentas acessíveis para uso na prática clínica. Métodos Revisão sistemática conduzida nos bancos de dados PubMed®, Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS), MEDLINE® e Web of Science utilizando os termos de pesquisa “endometriosis” (todos os campos) AND “miRNA” (todos os campos), avaliando todas as publicações até maio de 2019. Resultados A maioria dos miRNAs desregulados foram analisados a partir de amostras de tecido, o que inviabiliza seu uso como teste diagnóstico não invasivo. Todavia, 62 miRNAs foram identificados como diferencialmente expressos em amostras que poderiam ser usadas para o diagnóstico pouco invasivo de endometriose, como sangue, soro e plasma. Conclusão Apesar de todos esses candidatos, os trabalhos são exploratórios, realizados com números pequenos de amostras, sem miRNAs específicos validados para fins diagnósticos. Estudos envolvendo principalmente amostras biológicas, visando à pesquisa translacional, deveriam ser mais explorados. O desenvolvimento de grandes bancos de dados sobre amostras, bem como o uso de saliva e fluido vaginal para identificação de miRNAs, poderia servir como recursos essenciais para as barreiras atuais no diagnóstico da endometriose.

ABSTRACT Objective The aim of the study was to assess the evidence on miRNAs as biomarkers for the diagnosis of endometriosis, as well as to provide insights into the challenges and strategies associated with the use of these molecules as accessible tools in clinical practice. Methods Systematic review conducted on PubMed®, Latin American and Caribbean Health Sciences Literature (LILACS), MEDLINE® and Web of Science databases using the search terms endometriosis (all fields) AND miRNA (all fields), evaluating all publication up to May 2019. Results Most miRNAs found to be dysregulated in this study were harvested from tissue samples, which precludes their use as a non-invasive diagnostic test. However, differential expression of 62 miRNAs was reported in samples that may be used for non-invasive diagnosis of endometriosis, such as blood, serum and plasma. Conclusion Despite the identification of several candidates, studies are investigatory in nature and have been conducted with small number of samples. Also, no particular miRNA has been validated for diagnostic purposes so far. Studies based primarily on biological samples and applicable to translational research are warranted. Large databases comprising information on sample type and the use of saliva and vaginal fluid for miRNAs identification may prove essential to overcome current barriers to diagnosis of endometriosis.

RESUMO Objetivo Analisar e comparar a expressão de receptores do tipo Toll por células T reguladoras presentes no líquido peritoneal de pacientes com endometriose. Métodos Células T reguladoras foram isoladas do líquido peritoneal de mulheres com e sem endometriose, coletadas durante a cirurgia, e o RNAm foi extraído para análise da expressão de receptores do tipo Toll por reação em cadeia da polimerase com transcriptase reversa. Resultados Pacientes com endometriose apresentaram células T reguladoras expressando maior número e variedade de Toll por células quando comparadas com T reguladoras de pacientes do Grupo Controle. Receptores do tipo Toll-1 e receptores do tipo Toll-2 foram expressos em ambos os grupos. Todos os outros tipos de receptores Toll foram encontrados expressos apenas em células T reguladoras do grupo com endometriose. Conclusão Pacientes com endometriose apresentaram células T reguladoras peritoneais expressando vários tipos de receptores tipo Toll.

ABSTRACT Objective To analyze and compare the expression of Toll-like receptors by regulatory T cells present in the peritoneal fluid of patients with and without endometriosis. Methods Regulatory T cells were isolated from peritoneal fluid of women with and without endometriosis, collected during surgery, and mRNA was extracted for analysis of Toll-like receptors expression by reverse-transcriptase polymerase chain reaction. Results Patients with endometriosis presented regulatory T cells expressing a larger number and variety of Toll-like receptors when compared to regulatory T cells from patients in the Control Group. Toll-like receptor-1 and Toll-like receptor-2 in regulatory T cells were expressed in both groups. All other expressed Toll-like receptors types were only found in regulatory T cells from the Endometriosis Group. Conclusion Patients with endometriosis had peritoneal regulatory T cells expressing various Toll-like receptors types.

RESUMO Objetivo: Avaliar características clínicas e complicações em pacientes com endometriose intestinal submetidos ao tratamento hormonal. Métodos: Dados de prontuários de 238 pacientes com endometriose de retossigmoide tratadas entre maio de 2010 e maio de 2016 foram coletados para este estudo retrospectivo. Resultados: Durante o período de acompanhamento, 143 (60,1%) mulheres mantiveram tratamento clínico, enquanto 95 (39,9%) tiveram piora dos sintomas de dor ou aumento da lesão intestinal (grupo falha de tratamento clínico), sendo 54 submetidas ao tratamento cirúrgico. As mulheres no Grupo Tratamento Clínico eram mais velhas (40,5±5,1 anos versus 37,3±5,8 anos; p<0,0001) e tinham lesões intestinais menores (2,1±1,9 versus 3,1±2,2; p=0,008) em comparação ao grupo falha de tratamento clínico. Redução significativa e semelhante do escore de dor na dismenorreia, dor pélvica crônica, disquezia cíclica e disúria cíclica foi observada nos Grupos Tratamento Clínico e Cirúrgico. Dispareunia, no entato, teve uma redução maior no Grupo Cirurgia. A redução subjetiva dos sintomas dolorosos também foi semelhante entre os Grupos Clínico e Cirúrgico (100% versus 98,2%; p=0,18). O Grupo Tratamento Cirúrgico foi relacionado a uma maior taxa de complicações graves (9,2% versus 0,6%; p=0,001) em comparação ao Grupo Tratamento Clínico. Conclusão: Falha no tratamento clínico em pacientes com endometriose de retossigmoide foi observada em mulheres mais jovens que tinham lesões intestinais maiores. O tratamento clínico hormonal foi igualmente eficaz na melhora dos sintomas de dor, exceto dispareunia, em comparação ao tratamento cirúrgico em mulheres com endometriose intestinal, mas com menor taxa de complicações. O tratamento clínico deve ser oferecido como primeira opção em pacientes com endometriose intestinal, enquanto o tratamento cirúrgico deve ser reservado para pacientes sem melhora nos sintomas de dor com tratamento hormonal, progressão das lesões ou suspeita de suboclusão intestinal.

ABSTRACT Objective: To evaluate clinical features and complications in patients with bowel endometriosis submitted to hormonal therapy. Methods: Retrospective study based on data extracted from medical records of 238 women with recto-sigmoid endometriosis treated between May 2010 and May 2016. Results: Over the course of follow-up, 143 (60.1%) women remained in medical treatment while 95 (39.9%) presented with worsening of pain symptoms or intestinal lesion growth (failure of medical treatment group), with surgical resection performed in 54 cases. Women in the Medical Treatment Group were older (40.5±5.1 years versus 37.3±5.8 years; p<0.0001) and had smaller recto sigmoid lesions (2.1±1.9 versus 3.1±2.2; p=0.008) compared to those who had failed to respond to medical treatment. Similar significant reduction in pain scores for dysmenorrhea, chronic pelvic pain, cyclic dyschezia and dysuria was observed in both groups; however greater reduction in pain scores for dyspareunia was noted in the Surgical Group. Subjective improvement in pain symptoms was also similar between groups (100% versus 98.2%; p=0.18). Major complications rates were higher in the Surgical Group (9.2% versus 0.6%; p=0.001). Conclusion: Patients with recto-sigmoid endometriosis who failed to respond to medical treatment were younger and had larger intestinal lesions. Hormonal therapy was equally efficient in improving pain symptoms other than dyspareunia compared to surgery, and was associated with lower complication rates in women with recto-sigmoid endometriosis. Medical treatment should be offered as a first-line therapy for patients with bowel endometriosis. Surgical treatment should be reserved for patients with pain symptoms unresponsive to hormonal therapy, lesion growth or suspected intestinal subocclusion.

O ácido jasmônico e seus derivados jasmonatos são reguladores vegetais endógenos produzidos por várias espécies vegetais, que atuam no mecanismo de defesa das plantas e agem como sinalizadores de estresse. O objetivo deste trabalho foi comparar o potencial fitorregulador de um fermentado, obtido pela fermentação do fungo filamentoso Botryosphaeria rhodina, contendo jasmonatos, comparado à ação do ácido jasmônico (AJ) usado como padrão. Sementes selecionadas de Capsicum frutescens (pimenta malagueta) e Physalis angulata (camapú) foram plantadas em terra contendo fertilizantes orgânicos e minerais. Plantas após 37 dias de germinação foram tratadas via pulverização foliar com duas diferentes concentrações de AJ (25,0 e 50,0 mg L-1) e com fermentado contendo jasmonatos (correspondendo a 50 mg L-1 de AJ) e como controle foi utilizado água. Os experimentos foram realizados em casa de vegetação, em blocos casualizados, com três repetições de cinco plantas por tratamentos, com os tratamentos dispostos em fatorial de 4x1 (1 controle; 2 doses de AJ e 1 dose do filtrado). A avaliação das plantas foi realizada 80 dias após a aplicação de AJ e o fermentado. Plantas controle de Capsicum produziram 6,4 frutos/planta enquanto que plantas tratadas com 25,0 e 50,0 mg L-1 de AJ produziram 7,2 frutos/planta e 9,2 frutos/planta, respectivamente e plantas tratadas com o fermentado produziram 11,7 frutos/planta. Adicionalmente, plantas controle de Physalis produziram 10,5 botões/planta e quando tratadas com 25,0 e 50,0 mg L-1 de AJ produziram 15,5 e 16,0 botões/planta, respectivamente. Plantas tratadas com o fermentado dobraram a produção de botões/planta (23,0) quando comparados ao controle. Resultados obtidos validam o potencial fitorregulador do AJ e do fermentado contendo jasmonatos, comprovado pelo aumento na produção de frutos e botões florais pelas espécies vegetais testadas, evidenciando assim o potencial de uso deste fermentado como alternativa de menor custo ao AJ obtido comercialmente.

Jasmonic acid (JA) and its jasmonate derivatives are endogenous hormones produced by numerous plant species that interfere on plant defense mechanism and act as stress signaling. The aim of this work was to compare the plant growth regulator potential of a fermented broth, obtained by the fermentation of the filamentous fungus Botryosphaeria rhodina, containing jasmonates to the action of the jasmonic acid used as standard. Selected seeds of Capsicum frutescens (pimenta malagueta) and Physalis angulata (camapú) were planted in soil containing mineral and organic fertilizer. Thirty seven days after germination seedlings were sprinkled with two different concentrations of JA (25.0 and 50.0 mg L-1) and with the jasmonate-containing fermented broth (corresponding to 50 mg L-1 of JA). Water was used as control. The experiment was carried out in a greenhouse in a completely randomized block design, with three replications with five plants and treatments applied to a 4x1 factorial (1 control; 2 doses of JA and 1 dose of filtrated). The evaluation of seedlings was performed 80 days after scattering with JA and the fermented broth. Capsicum control plants produced 6.4 fruits/plant, while plants sprinkled with 25.0 and 50.0 mg L-1 JA produced 7.2 and 9. 2 fruits/plant respectively and plants sparged with the fermented broth produced 11.7 fruits/plant. Moreover, Physalis control plants produced 10.5 buds/plant and when sprinkled with 25.0 and 50.0 mg L-1 JA produced 15.5 and 16.0 buds/plant respectively. Plantlets sparged with the fermented broth doubled the production of buds/plant (23.0) when compared to the control. The results obtained validate the growth regulator potential of JA and of the fermented broth on the tested plants, evidenced by the increased production of fruits and flower buds by the plant species tested. Thus, the fermented broth of the fungus Botryosphaeria rhodina is a lower cost alternative to commercially obtained JA.