Developing an innovative 3D printing platform for production of personalised medicines in a hospital for the OPERA clinical trial

Breast cancer is the most frequently diagnosed cancer in women worldwide, and non-adherence to adjuvant hormonotherapy can negatively impact cancer recurrence and relapse. Non-adherence is associated with side effects of hormonotherapy. Pharmacological strategies to mitigate the side effects include coadministration of antidepressants, however patients remain non-adherent. The aim of this work was to develop medicines containing both hormonotherapy, tamoxifen (20 mg), along with anti-depressants, either venlafaxine (37.5 or 75 mg) or duloxetine (30 or 60 mg), to assess the acceptability and efficacy of this personalised approach for mitigating tamoxifen side effects in a clinical trial. A major criterion for the developed medicines was the production rate, specified at minimum 200 dosage units per hour to produce more than 40,000 units required for the clinical trial. A novel capsule filling approach enabled by the pharmaceutical 3D printer M3DIMAKER 2 was developed for this purpose. Firstly, semi-solid extrusion 3D printing enabled the filling of tamoxifen pharma-ink prepared according to French compounding regulation, followed by filling of commercial venlafaxine or duloxetine pellets enabled by the development of an innovative pellet dispensing printhead. The medicines were successfully developed and produced in the clinical pharmacy department of the cancer hospital Gustave Roussy, located in Paris, France. The developed medicines satisfied quality and production rate requirements and were stable for storage up to one year to cover the duration of the trial. This work demonstrates the feasibility of developing and producing combined tamoxifen medicines in a hospital setting through a pharmaceutical 3D printer to enable a clinical trial with a high medicines production rate requirement

Níveis de deoxinivalenol, compostos fenólicos e atividade antioxidante de farinha de trigo após processo de polimento

The objective of this work was to evaluate the effect of the debranning process on the deoxynivalenol (DON) content, phenolic compounds, and antioxidant activity of wheat flours, in order to determine which milling technology provides the safest and most nutritional wheat products. Grain samples from the BRS Marcante, BRS Reponte, and BRS 374 wheat (Triticum aestivum) cultivars were used to obtain both whole-wheat and white flours. For whole-wheat, grains were debranned at different times (20 and 40 s). The debranning process significantly reduced the DON content in the whole-wheat flours from all cultivars. The DON concentration decreased 22 and 28% in the 20 and 40 s debranning treatments, respectively, when compared with the treatment without debranning. In addition, phenolic compound content and antioxidant capacity are significantly higher in the whole-wheat flours. The debranning process contributes to the production of safer and healthier foods, by reducing DON content and retaining phenolic compounds and antioxidant activity in whole-wheat flours.O objetivo deste trabalho foi avaliar o efeito do processo de polimento nos níveis de deoxinivalenol (DON), nos compostos fenólicos e na atividade antioxidante de farinhas de trigo, para determinar a tecnologia de moagem que fornece os produtos de trigo mais seguros e nutritivos. Amostras de grãos das cultivares de trigo (Triticum aestivum) BRS Marcante, BRS Reponte e BRS 374 foram utilizadas para obter farinhas de trigo integral e branca. Para a farinha integral, os grãos foram submetidos a diferentes períodos (20 e 40 s) de polimento. O processo de polimento reduziu significativamente o teor de DON nas farinhas de trigo integral de todas as cultivares. A concentração de DON diminuiu 22 e 28% nos tratamentos polimento por 20 e 40 s, respectivamente, quando comparada à do tratamento sem polimento. Além disso, os níveis de compostos fenólicos e a capacidade antioxidante são significativamente maiores nas farinhas de trigo integral. O processo de polimento contribui para a produção de alimentos mais seguros e saudáveis, ao reduzir o teor de DON e conservar os compostos fenólicos e a atividade antioxidante na farinha de trigo integral

The use of cyclosporine modifies the clinical and histopathological presentation of tuberculosis after renal transplantation

Tuberculosis is one of the most frequent opportunistic infections after renal transplantation and occurred in 30 of 1264 patients transplanted between 1976 and 1996 at Hospital São Paulo - UNIFESP and Hospital Dom Silvério, Brazil. The incidence of 2.4% is five times higher than the Brazilian general population. The disease occurred between 50 days to 18 years after the transplant, and had an earlier and worse development in patients receiving azathioprine, prednisone and cyclosporine, with 35% presenting as a disseminated disease, while all patients receiving azathioprine and prednisone had exclusively pulmonary disease. Ninety percent of those patients had fever as the major initial clinical manifestation. Diagnosis was made by biopsy of the lesion (50%), positivity to M. tuberculosis in the sputum (30%) and spinal cerebral fluid analysis (7%). Duration of treatment ranged from 6 to 13 months and hepatotoxicity occurred in 3 patients. The patients who died had a significant greater number of rejection episodes and received higher doses of corticosteroid. In conclusion, the administration of cyclosporine changed the clinical and histopathological pattern of tuberculosis occurring after renal transplantation.A tuberculose é uma das mais frequentes infecções oportunistas encontradas após o transplante renal. A tuberculose foi diagnosticada em 30 de 1.264 pacientes que foram transplantados entre 1976 e 1996 na unidade de Transplante Renal do Hospital São Paulo - UNIFESP e no Hospital Dom Silvério Gomes Pimenta em São Paulo. A incidência de 2,4%, é cinco vezes superior à estimada para população geral brasileira. O diagnóstico foi realizado entre 50 dias a 18 anos após o transplante. Nos pacientes com esquema duplo de imunossupressão (prednisona e azatioprina), a manifestação inicial foi mais tardia, sendo que em 11 dos 13 pacientes ocorreu após os primeiros 3 anos de transplante. Enquanto com esquema tríplice (prednisona, azatioprina e ciclosporina), em 14 de 17 pacientes a doença ocorreu dentro dos primeiros 3 anos. Com esquema duplo de imunossupressão não houve manifestação de doença extra-torácica, em contrapartida com esquema tríplice de imunossupressão a forma de apresentação disseminada ocorreu em 35% dos casos, sendo mais precoce e mais grave. A febre foi o sintoma clínico principal em 90% dos casos. O diagnóstico foi confirmado, em 50% (n=15) através da biópsia dos órgãos envolvidos, em 30% (n=9) através da pesquisa de bacilos álcool ácido resistentes (BAAR) no escarro, em 7% (n=2) no líquido céfalo-raquidiano e em 13% (n=4) através de necrópsia. O tempo de tratamento da tuberculose variou de 6 a 13 meses. A hepatotoxidade relacionada ao tratamento, ocorreu em três casos. O número de tratamentos prévios de episódios de rejeição e a dose administrada de corticóides foram maiores nos pacientes que morreram com este diagnóstico, quando comparados ao grupo que respondeu ao tratamento. Em conclusão a forma clínica e histopatológica de apresentação da tuberculose é diferente após o transplante em pacientes utilizando ciclosporina, sendo a resposta ao tratamento influenciada pela dose prévia de corticóide

Molecular characterization of Cyclophilin (TcCyP19) in Trypanosoma cruzi populations susceptible and resistant to benznidazole

Cyclophilin (TcCyP19), a peptidyl-prolyl cis/trans isomerase, is a key molecule with diverse biological functions that include roles in molecular chaperoning, stress response, immune modulation, and signal transduction. In this respect, TcCyP19 could serve as a potential drug target in diseasecausing parasites. Previous studies employing proteomics techniques have shown that the TcCyP19 isoform was more abundant in a benznidazole (BZ)-resistant Trypanosoma cruzi population than in its susceptible counterpart. In this study, TcCyP19 has been characterized in BZ-susceptible and BZresistant T. cruzi populations. Phylogenetic analysis revealed a clear dichotomy between Cyphophilin A (CyPA) sequences from trypanosomatids and mammals. Sequencing analysis revealed that the amino acid sequences of TcCyP19 were identical among the T. cruzi samples analyzed. Southern blot analysis showed that TcCyP19 is a single-copy gene, located in chromosomal bands varying in size from 0.68 to 2.2 Mb, depending on the strain of T. cruzi. Northern blot and qPCR indicated that the levels of TcCyP19 mRNA were two-fold higher in drug-resistant T. cruzi populations than in their drugsusceptible counterparts. Similarly, as determined by two-dimensional gel electrophoresis immunoblot, the expression of TcCyP19 protein was increased to the same degree in BZ-resistant T. cruzi populations. No differences in TcCyP19 mRNA and protein expression levels were observed between the susceptible and the naturally resistant T. cruzi strains analyzed. Taken together, these data indicate that cyclophilin TcCyP19 expression is up-regulated at both transcriptional and translational levels in T. cruzi populations that were in vitro-induced and in vivo-selected for resistance to BZ.Fil: Rêgo, Juciane Vaz. Universidade Federal Do Piaui.; BrasilFil: Duarte, Ana Paula. Fundación Oswaldo Cruz; BrasilFil: Liarte, Daniel Barbosa. Universidade Federal Do Piaui.; BrasilFil: de Carvalho Sousa, Francirlene. Universidade Federal Do Piaui.; BrasilFil: Barreto, Humberto Medeiros. Universidade Federal Do Piaui.; BrasilFil: Bua, Jacqueline Elena. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”. Instituto Nacional de Parasitología "Dr. Mario Fatala Chaben”; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Romanha, Alvaro José. Fundación Oswaldo Cruz; BrasilFil: Rádis Baptista, Gandhi. Universidade Federal Do Piaui.; BrasilFil: Murta, Silvane Maria Fonseca. Universidade Federal Do Piaui.; Brasi

Análise da missão de fornecedores da indústria automotiva

O presente trabalho teve como objetivo analisar a missão de empresas fornecedoras do ramo automotivo instaladas no Brasil com relação aos aspectos definidos na literatura. Adotou-se uma abordagem qualitativa e quantitativa com base na pesquisa documental em dados secundários divulgados na internet e pesquisa bibliográfica para o referencial teórico e fundamentação dos dados. Foram consultadas 167 empresas das quais somente 49 divulgam sua missão na internet. Os resultados mostraram que nem todas as missões das empresas analisadas atendem aos aspectos definidos na literatura. Estes resultados podem ocorrer pelo desconhecimento dos itens que devem constar em uma missão ou por as empresas não darem à devida importância a esta questão

Tomografia computadorizada aplicada a estudos de um Planossolo

In order to find better techniques to evaluate the soil physical parameters applied to lowland soils, the computerized tomography has been used to measure soil density and water content with accuracy and high spatial resolution. This work was carried out in order to describe features and calibration procedures of a computerized minitomographer using X-ray and gama-rays as sources of radiation and to establish suitable statistical parameters on the study of soil bulk density and water content in a Planosol (Albaqualf) from Rio Grande do Sul State, Brazil. The minitomographer calibration was obtained from the linear regression equation among the tomography's unities (TU) presented by the image reconstruction program and the linear attenuation coefficient (ml, cm-1), by the measurement of direct transmission of g-rays as source of radiation in soil samples from A and B horizons, distilled water, benzin and aluminum. In order to get measures of the direct radiation transmission, containers with distilled water, benzin, soil and aluminum were used to obtain the following equations to calculate soil bulk density in the A horizon: Ds = [(TU/986.16)-(0.200xq)]/0.267 and in the B horizon: Ds = [(TU/986.16)-(0.200xq)]/0.297, where TU is the mean value in the line and q is the soil volumetric water content (m3 m-3). The obtained configurations allowed to attain average variabilities of 2.74% and 0.73% for homogeneity and repeatability, respectively. The expected errors related to the equipment are 0.051 and 0.046 Mg m-3, to the A and B horizons, respectively. The results showed the technique accuracy and adaptability in the studies of the physical characteristics of a Planosol.Na busca de técnicas mais apuradas para a determinação e avaliação de parâmetros físicos do solo com aplicabilidade em várzeas, vem se destacando a tomografia computadorizada, por medir a densidade e a umidade com boa sensibilidade e alta resolução espacial. O presente trabalho teve como objetivo descrever aspectos e procedimentos da calibração de um minitomógrafo de raios-X e gama para estudo da densidade e umidade de um Planossolo no Rio Grande do Sul, bem como estabelecer parâmetros estatísticos para sua adequada utilização. A calibração do minitomógrafo foi obtida pela regressão linear entre as unidades tomográficas (UT), apresentadas pelo programa de reconstrução de imagem, e os coeficientes de atenuação linear (μ1, cm-1), medidos por transmissão direta de raios gama, em amostras dos horizontes A e B do Planossolo, água destilada, benzina e alumínio. Para as medidas de transmissão direta de radiação utilizaram-se recipientes com água destilada, benzina, solo e Al, obtendo-se as seguintes fórmulas para o cálculo da densidade do solo no horizonte A: Ds = [(UT/986,16)-(0,200xθ)]/0,267; e no horizonte B: Ds = [(UT/986,16)-(0,200xθ)]/0,297, em que UT é o valor médio de UT em cada linha e θé a umidade volumétrica da amostra de solo, em m3 m-3. Com as configurações obtidas, verificou-se variabilidade média de 2,74% e 0,73%, respectivamente, em termos de homogeneidade e repetibilidade. Os erros atribuídos ao equipamento são de 0,051 e 0,046 Mg m-3, respectivamente, nos horizontes A e B, revelando precisão e adaptabilidade no emprego da técnica em estudos do Planossolo

CD137 (4-1BB) requires physically associated cIAPs for signal transduction and antitumor effects

CD137 (4-1BB) is a member of the TNFR family that mediates potent T cell costimulatory signals upon ligation by CD137L or agonist monoclonal antibodies (mAbs). CD137 agonists attain immunotherapeutic antitumor effects in cancer mouse models, and multiple agents of this kind are undergoing clinical trials. We show that cIAP1 and cIAP2 are physically associated with the CD137 signaling complex. Moreover, cIAPs are required for CD137 signaling toward the NF-κB and MAPK pathways and for costimulation of human and mouse T lymphocytes. Functional evidence was substantiated with SMAC mimetics that trigger cIAP degradation and by transfecting cIAP dominant-negative variants. Antitumor effects of agonist anti-CD137 mAbs are critically dependent on the integrity of cIAPs in cancer mouse models, and cIAPs are also required for signaling from CARs encompassing CD137’s cytoplasmic tail.I.M. has been granted with PID2020-112892RB funded by MICIN/AEI/10.13039/501100011033 and SAF2017-83267-C2-1-R funded by MICIN/AEI/10.13039/501100011033/ and by FEDER “Una manera de hacer Europa,” (HR21-00083) the Fundación La Caixa, “MINCITH. Metabolic requirements for immune INfiltration in effective Cancer ImmunoTHerapy” “AYUDAS FUNDACIÓN BBVA A EQUIPOS DE INVESTIGACIÓN CIENTÍFICA 2019” Fundación BBVA, the Instituto de Salud Carlos III (PI20/00002 and PI19/01128), cofinanced by the Fondos FEDER “A way to make Europe” and Joint Translational Call for Proposals 2015 (JTC 2015), TRANSCAN456 2 (code TRS-2016-00000371), and the Gobierno de Navarra Proyecto LINTERNA (reference 0011-1411-2020-000075). Funding was also received from B. J. Baselga (Fundación FERO) and the T2-EVOLVE project from the EU. I.M. and M.A. receive grant funding from Pharmamar and Highlight Therapeutics. M.A. is supported by AECC (INVES1904ALVA). J.M.Z. has been granted with PID2019-110405RB-I00 funded by MICIN/AEI/10.13039/501100011033 and with P2022/BMD-7225 funded by Consortium in Biomedicine of Comunidad de Madrid.Peer reviewe

mRNAs encoding IL-12 and a decoy-resistant variant of IL-18 synergize to engineer T cells for efficacious intratumoral adoptive immunotherapy

Interleukin-12 (IL-12) gene transfer enhances the therapeutic potency of adoptive T cell therapies. We previously reported that transient engineering of tumor-specific CD8 T cells with IL-12 mRNA enhanced their systemic therapeutic efficacy when delivered intratumorally. Here, we mix T cells engineered with mRNAs to express either single-chain IL-12 (scIL-12) or an IL-18 decoy-resistant variant (DRIL18) that is not functionally hampered by IL-18 binding protein (IL-18BP). These mRNA-engineered T cell mixtures are repeatedly injected into mouse tumors. Pmel-1 T cell receptor (TCR)-transgenic T cells electroporated with scIL-12 or DRIL18 mRNAs exert powerful therapeutic effects in local and distant melanoma lesions. These effects are associated with T cell metabolic fitness, enhanced miR-155 control on immunosuppressive target genes, enhanced expression of various cytokines, and changes in the glycosylation profile of surface proteins, enabling adhesiveness to E-selectin. Efficacy of this intratumoral immunotherapeutic strategy is recapitulated in cultures of tumor-infiltrating lymphocytes (TILs) and chimeric antigen receptor (CAR) T cells on IL-12 and DRIL18 mRNA electroporation.Fil: Olivera, Irene. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Bolaños, Elixabet. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Gonzalez Gomariz, Jose. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Hervas Stubbs, Sandra. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Mariño, Karina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Luri Rey, Carlos. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Etxeberria, Iñaki. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Cirella, Assunta. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Egea, Josune. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Glez Vaz, Javier. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Garasa, Saray. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; España. Centro de Investigación Biomédica en Red de Cáncer ; EspañaFil: Alvarez, Maite. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; España. Centro de Investigación Biomédica en Red de Cáncer ; EspañaFil: Eguren Santamaria, Iñaki. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Guedan, Sonia. Institut D'investigacions Biomèdiques August Pi Isunyer; EspañaFil: Sanmamed, Miguel F.. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; España. Centro de Investigación Biomédica en Red de Cáncer ; EspañaFil: Berraondo, Pedro. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; España. Centro de Investigación Biomédica en Red de Cáncer ; EspañaFil: Rabinovich, Gabriel Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaFil: Teijeira, Alvaro. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; EspañaFil: Melero, Ignacio. Universidad de Navarra; España. Instituto de Investigación Sanitaria de Navarra; España. Centro de Investigación Biomédica en Red de Cáncer ; España. Clínica Universidad de Navarra; Españ

Tamanho de amostras para a determinação de parâmetros físicos em planossolo por tomografia computadorizada

A técnica da tomografia computadorizada (TC) permite medir a densidade e a umidade de amostras de solo, constituindo uma importante ferramenta na Ciência do Solo. Este trabalho tem como objetivos descrever os aspectos da adequação do tamanho de amostras de um Planossolo e os procedimentos de avaliação e estudos por análise estatística, empregando-se um minitomógrafo computadorizado de raios gama com fonte de 241Am. O valor do erro atribuído ao equipamento são 0,051 e 0,046 Mg m-3, respectivamente, para os horizontes A e B. O valor teórico da espessura da amostra do Planossolo para uso na técnica de TC com fonte de 241Am é, aproximadamente, 4,0 cm para os horizontes A e B. Já a espessura ideal de amostras é de aproximadamente 6,0 cm, sendo menor para amostras do horizonte B em relação ao A. Obteve-se boa precisão e adaptabilidade no emprego da TC para estudos de Planossolos._________________________________________________________________________________ ABSTRACT: Computerized tomography (CT) is an important tool in Soil Science for noninvasive measurement of density and water content of soil samples. This work aims to describe the aspects of sample size adequacy for Planosol (Albaqualf) and to evaluate procedures for statistical analysis, using a CT scanner with a 241Am source. Density errors attributed to the equipment are 0.051 and 0.046 Mg m-3 for horizons A and B, respectively. The theoretical value for sample thickness for the Planosol, using this equipment, is 4.0 cm for the horizons A and B. The ideal thickness of samples is approximately 6.0 cm, being smaller for samples of the horizon B in relation to A. Alternatives for the improvement of the efficiency analysis and the reliability of the results obtained by CT are also discussed, and indicate good precision and adaptability of the application of this technology in Planosol (Albaqualf) studies