REPOSITÓRIO INSTITUCIONAL DA UNIVERSIDADE DO VALE DO PARAÍBA
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Submissões Recentes
Cost-effective production process of scFv antibody fragments against Shiga toxin 2 via recombinant E. coli
(Elsevier) Guimarães, Marcela; Luz, Daniela; Augusto, Elisabeth de Fátima Pires; Vieira, Lucia; Costa, Maricilia Silva; Piazza, Roxane Maria Fontes; Pradella, José Geraldo da Cruz
Shiga toxin (Stx)-producing Escherichia coli (STEC) and its subgroup enterohemorrhagic E. coli are significant pathogens responsible for diarrhea, which can progress to hemorrhagic colitis and hemolytic uremic syndrome (HUS), the leading cause of acute renal failure in children. Early diagnosis is crucial for effective clinical man- agement, as antibiotic treatment is not recommended for STEC infections. The present study aimed to establish a cost-effective biotechnological platform for cultivating recombinant E. coli to produce scFv antibody fragments against Stx2 for diagnostic applications. The method was first evaluated through shake flask experiments and subsequently scaled up to bench-scale bioreactors operated in both batch and fed-batch modes using defined culture media. Optimal production conditions were achieved by inducing recombinant E. coli pLys at 18 ◦C for 18 h with 0.1 mM IPTG, resulting in a yield of 3.0 to 4.0 mg scFv/g cell biomass. A fed-batch, high-cell-density procedure with E. coli pLysS achieved a maximum production up to 150 mg scFv/L. A preliminary economic assessment demonstrated the production potential at a value of around $250/g scFv. Economic analysis also highlights that the relative cost of capital investment becomes important as production processes intensify. Therefore, technical parameters such as productivity (scFv mass/bioreactor volume * time) and scFv concentration (mass scFv mass/bioreactor volume) should be prioritized to maximize their values. Similarly, optimization of the recombinant E. coli microbial platform should be pursued to increase the Yp/x level.
A microbead-enhanced electrochemical platform for β-amyloid peptide (1–42) detection
(Frontiersin) Razzino, Claudia do Amaral; Sgobbi, Lívia Flório; Cancino-Bernardi, Juliana; Zapata, Angelica Maria Mazuera; Costa, Clara Cardoso; Zucolotto, Valtencir; Vieira, Lucia; Lobo, Anderson Oliveira
Alzheimer’s disease is the most prevalent form of dementia and is primarily characterized by the accumulation of β-amyloid and phosphorylated tau proteins in the brain, along with the degeneration of nerve cells, which leads to impairment of various cognitive functions. A significant biomarker of Alzheimer’s disease is the decreased level of soluble β-amyloid peptide (1–42) (Aβ1-42) in cerebrospinal fluid (CSF), as pathology progresses when CSF-Aβ1-42 levels drop below 192 pg mL-1 . In this study, we developed an amperometric immunosensor based on magnetic beads as the platform for constructing the immunosensor. Monoclonal antibodies are immobilized on the MBs, enabling selective detection of Aβ1-42. The detection antibody is conjugated with the enzyme horseradish peroxidase, which, in the presence of H2O2 and hydroquinone, catalyzes the decomposition of H2O2 and the oxidation of hydroquinone to p-quinone, generating an electric current measured at a potential of −200 mV (vs. the Ag pseudo-reference electrode) using screen-printed carbon electrodes. The amperometric sandwich-type immunosensor demonstrates a linear response
in the concentration range of 10 to 10,000 pg mL-1 , with a detection limit of 7.4 pg mL-1 , exhibiting excellent selectivity against the assessed interferents. These findings suggest the potential application of this immunosensor in the early diagnosis of Alzheimer’s disease, offering a sensitive and specific tool for clinical analysis. Despite its high performance, further studies are required to validate its robustness and applicability in complex clinical samples.
Tribocorrosion Susceptibility and Cell Viability Study of 316L Stainless Steel and Ti6Al4V Titanium Alloy with and without DLC Coatings
(MDPI) Sene, Ana Claudia; Silva, Michely Glenda Pereira da; Macário, Paulo Fabrício; Vieira, Angela Aparecida; Leite, Priscila Maria Sarmeiro Correa Marciano; Silva, Newton Soares da; Marques, Francisco Das Chagas; Vieira, Lúcia
Stainless steel (SS316L) and titanium alloy (Ti6Al4V) exhibit suitable properties for biomed- ical applications; however, the tribocorrosion of these materials, which is associated with metallosis, is still a significant concern. This work investigates the effectiveness of DLC smoothing coatings applied to the metals to reduce tribocorrosion and improve cell viability. The study was motivated by many reports of metallosis caused by metal debris in the soft tissues of the body. DLC coatings were produced using the plasma-enhanced chemical vapor deposition (PECVD) technique. The cytotoxicity, genotoxicity, and cell viability of metallic samples with and without DLC coatings were analyzed, considering the chemical composition of the coating and metallic components. The results show that the DLC coatings presented suitable interaction properties and no cytotoxicity or genotoxicity when exposed to the cellular environment, compared with the control group (p < 0.0001). They also demonstrated cell viability, low friction representing a reduction of 80%, and hardness 23–26 GPa, making them ideal for use on fixed implants. It is necessary to control the thickness and roughness of the coating to avoid pinholes and increase the corrosion protection of implants. These DLC coatings with low friction coefficients could facilitate the fixation of implantable pins and screws, including Kirschner wires.
Nano-Enabled Colorimetric Assay for the Detection of Paracoccidioides lutzii: Advancing Diagnostics with Nanotechnology
(MDPI) Comparato Filho, Olavo de Osti; Cândido, Marcela Aparecida; Ventura, Aveline; Morais, Flavia Villaça; Raniero, Leandro José
Deforestation is a common occurrence driven by agricultural expansion, urbanization, and infrastructure development. These activities often lead to increased human interaction with ecosys- tems, potentially exposing individuals to Paracoccidioides spores (P. brasiliensis and P. lutzii) found in the soil, resulting in Paracoccidioidomycosis (PCM). This fungal infection is endemic to specific regions in Latin America, such as Brazil, Colombia, Venezuela, and Argentina. Diagnosis typically involves a combination of clinical assessment, imaging techniques, and laboratory examinations. P. lutzii lacks the glycoprotein Gp43, a key antigenic protein utilized in serological tests for PCM diagnosis. In this study, a colorimetric test employing gold nanoparticles (AuNPs) and label-free methodology was employed for P. lutzii detection. The effectiveness of the label-free colorimetric test was assessed using a total of 100 samples. This detection was achieved through the amplification of the gp43 gene and the use of a specific probe (5′CAGGGGTGCG3′) in conjunction with AuNPs. The receiver operating characteristic curve was employed to assess the test, revealing that the method can accurately detect P. lutzii with a sensitivity of 100% and a specificity of 100%. The findings indicate a substantial impact on remote endemic regions attributable to the implementation of cost-effective diagnostic methodologies.
Stepwise Nitric Oxide Release and Antibacterial Activity of a Nitric Oxide Photodonor Hosted within Cyclodextrin Branched Polymer Nanocarriers
(ACS Publications) Martins, Tassia Joi; Parisi, Cristina; Pinto, Juliana Guerra; Brambilla, Isabelle de Paula Ribeiro; Malanga, Milo; Ferreira-Strixino, Juliana; Sortino, Salvatore
A hydrophobic nitric oxide (NO) photodonor integrating both nitroso and nitro functionalities within its chromophoric skeleton has been synthesized. Excitation of this compound with blue light triggers the release of two NO molecules from the nitroso and the nitro functionalities via a stepwise mechanism. Encapsulation of the NO photodonor within biocompatible neutral, cationic, and anionic β-cyclodextrin branched polymers as suitable carriers leads to supramolecular nanoassemblies, which exhibit the same nature of the photochemical processes but NO photorelease performances enhanced by about 1 order of magnitude when compared with the free guest. Antibacterial tests carried out with methicillin-resistant Staphylococcus aureus and Acinetobacter baumannii demonstrate an effective antibacterial activity exclusively under light activation and point out a differentiated role of the polymeric nanocarriers in determining the outcome of the antibacterial photodynamic action.