REPOSITÓRIO INSTITUCIONAL DA UNIVERSIDADE DO VALE DO PARAÍBA
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Submissões Recentes
3D bioprinted human iPSC-derived neural progenitor cells as a novel platform for studying neurogenic niche
(AIP Publishing) Machado, Lucas Simões; Ferreira, Paula Scanavez; Pires, Marina Rodrigues; Bim, Larissa Valdemarin; Oliveira, Natália Heloísa de; Salles, Geisa Rodrigues; Ferreira, Natalia Dall'Agnol; Cruz, Elisa Marozzi; Porcionatto, Marimelia Aparecida
Animal models, especially rodents, used to study neurodevelopment have significantly advanced our comprehension of cellular and molecular mechanisms. Nevertheless, differences in species-specific structures, gestation periods, and interneuronal connections limit animal models’ ability to represent human neurodevelopment accurately. The unique characteristics of primate neural progenitor cells (NPCs) enable cortex expansion with gyrus formation, which does not occur in lissencephalic animals, like rodents. Therefore, there is a need for novel in vitro models using human cells that recapitulate the complexity of human brain development. Along with organoids, 3D bioprinting offers a platform for creating more complex in vitro models. We developed, extensively characterized, and successfully used a GeltrexTM/GelMA hydrogel blend to bioprint human induced pluripotent stem cells-derived NPCs (hNPCs). We show that 3D bioprinted hNPCs can selforganize, revealing key features of a neurogenic niche, including proliferation, differentiation, and migration, remaining viable for over 110 days. Within the first 20 days, bioprinted constructs showed the formation of positive cell clusters for the neurogenic niche cell markers FABP7, NESTIN, and GFAP. Clusters were interconnected by process bundles supporting cell migration. The cells proliferated within the clusters, and over time, NPCs originated TUBB3þ neurons with long axonal tracts, prominent around the clusters. We propose this as a 4D model to study neurogenic niches’ key cellular and molecular features in a 3D bioprinted scaffold, adding time as the fourth dimension. Neuronal maturation in this dynamic model recapitulates key neurogenic niche properties, making it suitable for neurodevelopmental disease modeling and drug screening.
The arrangement of dual-species biofilms of Candida albicans and Issatchenkia orientalis can be modified by the medium: effect of Voriconazole
(Taylor & Francis) Passos, Juliene Cristina da Silva; Rodrigues, Ana Beatriz Furtado; Silva, Carlos Alberto; Costa, Maricilia Silva
Both Candida albicans and Issatchenkia orientalis have been isolated from different types of infections over the years. They have the ability to form communities of microorganisms known as biofilms. It has been demonstrated that the medium employed in studies may affect the biofilm development. The aim of this study was to investigate the arrangement of dual-species biofilms of C. albicans and I. orientalis cultivated on either RPMI-1640 or Sabouraud Dextrose Broth (SDB), as well as the inhibitory effect of Voriconazole (VRC). For the experiments performed, ATCC strains were used, and yeast-mixed suspensions were inoculated in 96-well plates with either RPMI-1640 or SDB, in the presence or absence of VRC. The results were observed by counting the number of CFU obtained from scraping off the biofilms produced and plating the content on CHROMagar Candida medium. It was observed that for all conditions tested the medium chosen affected the arrangement of dual-species biofilms: when RPMI-1640 was used, there was a prevalence of C. albicans, while the opposite was noted when SDB was used. It could be suggested that the medium and environment could regulate interactions between both yeast species, including the response to different antifungal drugs.
In-situ transmission electron microscopy (TEM) investigation of the reduction process in graphene oxide
(Springer Nature Link) Silva, Douglas Soares da; Viana, Gustavo Alexandre; Silva Filho, José Maria Clemente da; Kretly, Luiz Carlos; Chaves Neto, Antônio Maia de Jesus; Vieira, Lúcia; Barros, Tárcio André dos Santos; Marques, Francisco das Chagas
The reduction processes of graphene oxide (GO) aim to remove functional groups such as H2O, CO, and CO2 to promote the properties of GO towards those of pure graphene. We adopted the thermal reduction process from room temperature to 320 °C. The transmission electron microscopy (TEM) technique was used to probe the effect of reduction mechanisms. It was observed that the plasmon peaks, referring to sp2 carbon bonds in crystalline structures, are more evident in the reduced graphene oxide (rGO) than GO. The fine structure at the K edge of carbon shows differences in shape linked to the density of states above the Fermi level. Electron energy loss spectroscopy (EELS) analyses revealed an increase in the fraction of sp3 bonds in the reduced sample, consistent with the reduction of functional radicals in the GO structure.
From the gut to the heart: probiotic therapy with Saccharomyces boulardii and its potential role on diabetic cardiomyopathy in a murine model
(Taylor & Francis) Brandão, Ana Beatriz Pereira; Albuquerque, Raquel Cristina Melo Ferreira de; Abreu, Isabel Cristina Mallosto Emerich de; Ferreira, Fabiana Gomes; Santos, Leticia Barssotti dos; Jensen, Leonardo; Souza, Leandro Eziquiel de; Ferreira, Sarah Gomes; Souza, Lívia Bruni de; Arisawa, Emilia Angela Lo Schiavo; Sant′Anna, Luciana Barros; Higa, Elisa Mieko Suemitsu; Casali, Adenauer Girardi; Aimbire, Flávio; Irigoyen, Maria Claudia Costa; Casali, Karina Rabello; Cunha, Tatiana Sousa
Background: We investigated whether the probiotic yeast Saccharomyces boulardii confers cardiometabolic protection and prevents diabetic cardiomyopathy by modulating inflammation, cardiac remodelling, cardiovascular function, and autonomic regulation.
Methods: Male C57BL/6 mice were allocated into four groups: Control (C), Diabetes (DM), Control+Saccharomyces boulardii (CSb), and Diabetes+Saccharomyces boulardii (DMSb). Diabetes was induced with intraperitoneal streptozotocin (STZ), and treatments (sterile water or Saccharomyces boulardii) were administered orally for 8weeks. Blood glucose, cytokines, and nitric oxide levels were measured, along with cardiac function via echocardiography and direct blood pressure recordings.
Results: Saccharomyces boulardii reduced blood glucose and increased cardiac IL-10 in diabetic mice, restoring nitric oxide levels. These effects were associated to reduced collagen deposition, preventing vascular damage and ventricular fibrosis, and were accompanied by improved systolic/ diastolic function and autonomic control.
Conclusion: Saccharomyces boulardii improved cardiac structure, function, and autonomic control in diabetic mice, supporting its potential as adjunct therapy for diabetic cardiomyopathy.
Targeted optimization of single-chain variable fragment (scFv) expression in E. coli using a design-of-experiment approach
(Elsevier) Guimaraes, Marcela; Carvalho, Rafaela Vieira; Luz, Daniela; Guilherme, Ariela Pedro Bom; Campos, Claudia Barbosa Ladeira de; Piazza, Roxane Maria Fontes; Pradella, José Geraldo da Cruz
Infections by Shiga toxin-producing Escherichia coli (STEC) pose a serious global health risk, potentially leading to hemolytic uremic syndrome (HUS), a life-threatening condition. This study aimed to establish an efficient expression system to produce an anti-Stx2 single-chain variable fragment (scFv) using three E. coli strains: BL21 (DE3), BL21(DE3) pLysS, and ArcticExpress (DE3). Initial tests showed that BL21(DE3) pLysS and ArcticExpress (DE3) produced 0.3–0.4 mg scFv/L using defined media and induction with 0.1 mM IPTG or 10 g/L lactose. A Plackett-Burman design was then used to optimize the concentrations of 2xYT medium, IPTG, and lactose during induction. Under these conditions, ArcticExpress (DE3) reached 19 mg/L of scFv when induced with 1.0 mM IPTG and 2.5 g/L 2xYT. In contrast, BL21(DE3) pLysS yielded 34 mg/L with 1.0 mM IPTG, 0.4 g/L 2xYT, and 0.2 g/L lactose, while IPTG alone resulted in 26 mg/L. The protein yield (Yp/x) also increased from 11.9 to 15.5 mg scFv/g cell. Thus, E. coli BL21(DE3) pLysS showed the highest potential for producing anti-Stx2 scFv, particularly under optimized induction conditions involving IPTG, 2xYT, and lactose. These findings highlight the potential of recombinant E. coli BL21(DE3) pLysS as a robust platform for the scalable production of functional anti-Stx2 scFv fragments. By enabling higher yields under optimized induction strategies, this approach provides an important basis for the development of low-cost diagnostic tools targeting STEC infections, which remain a critical global health challenge.