Characterization of Biodegradable Polymers for Porous Structure: Further Steps toward Sustainable Plastics.

Plastic pollution poses a significant environmental challenge, necessitating the investigation of bioplastics with reduced end-of-life impact. This study systematically characterizes four promising bioplastics-polybutylene adipate terephthalate (PBAT), polybutylene succinate (PBS), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), and polylactic acid (PLA). Through a comprehensive analysis of their chemical, thermal, and mechanical properties, we elucidate their structural intricacies, processing behaviors, and potential morphologies. Employing an environmentally friendly process utilizing supercritical carbon dioxide, we successfully produced porous materials with microcellular structures. PBAT, PBS, and PLA exhibit closed-cell morphologies, while PHBV presents open cells, reflecting their distinct overall properties. Notably, PBAT foam demonstrated an average porous area of 1030.86 µm2, PBS showed an average porous area of 673 µm2, PHBV displayed open pores with an average area of 116.6 µm2, and PLA exhibited an average porous area of 620 µm2. Despite the intricacies involved in correlating morphology with material properties, the observed variations in pore area sizes align with the findings from chemical, thermal, and mechanical characterization. This alignment enhances our understanding of the morphological characteristics of each sample. Therefore, here, we report an advancement and comprehensive research in bioplastics, offering deeper insights into their properties and potential morphologies with an easy sustainable foaming process. The alignment of the process with sustainability principles, coupled with the unique features of each polymer, positions them as environmentally conscious and versatile materials for a range of applications.

DNA-Encoded Multivalent Display of Chemically Modified Protein Tetramers on Phage: Synthesis and in Vivo Applications.

Phage display links the phenotype of displayed polypeptides with the DNA sequence in the phage genome and offers a universal method for the discovery of proteins with novel properties. However, the display of large multisubunit proteins on phages remains a challenge. A majority of protein display systems are based on monovalent phagemid constructs, but methods for the robust display of multiple copies of large proteins are scarce. Here, we describe a DNA-encoded display of a ∼ 200 kDa tetrameric l-asparaginase protein on M13 and fd phages produced by ligation of SpyCatcher-Asparaginase fusion (ScA) and PEGylated-ScA (PEG-ScA) to barcoded phage clones displaying SpyTag peptide. Starting from the SpyTag display on p3 or p8 coat proteins yielded constructs with five copies of ScA displayed on p3 (ScA-p3), ∼100 copies of ScA on p8 protein (ScA-p8) and ∼300 copies of PEG-ScA on p8 protein (PEG-ScA-p8). Display constructs of different valencies and chemical modifications on protein (e.g., PEGylation) can be injected into mice and analyzed by deep sequencing of the DNA barcodes associated with phage clones. In these multiplexed studies, we observed a density and protein-dependent clearance rate in vivo. Our observations link the absence of PEGylation and increase in density of the displayed protein with the increased rate of the endocytosis by cells in vivo. In conclusion, we demonstrate that a multivalent display of l-asparaginase on phages could be used to study the circulation life of this protein in vivo, and such an approach opens the possibility to use DNA sequencing to investigate multiplexed libraries of other multisubunit proteins in vivo.

Development of a cell-free protein synthesis protocol to rapidly screen L-asparaginase proteoforms by enzymatic activity

Abstract BACKGROUND Cell-free protein synthesis (CFPS) technology has emerged as a powerful tool for a variety of biotechnological applications, including the expression of different classes of biopharmaceutical products. L-Asparaginase (E.C. Number: 3.5.1.1, L-asparagine amidohydrolase) (L-ASNase) is an important biopharmaceutical used to treat leukemia, but expression of multiple proteoforms in CFPS systems and rapid characterization using standard colorimetric methods has not yet been fully exploited. Herein, recombinant expression and characterization of an L-ASNase from Erwinia chrysanthemi (Erwinase) using a new CFPS protocol is reported. RESULTS Expression and quantification of the enzymatic activity of a soluble his-tagged L-ASNase directly from a CFPS reaction was successfully achieved. Purification of the protein was not required in order to assess its biological activity. Activity of L-ASNase was significantly higher than the control reaction (7.07 ± 0.68 U mL–1 vs. 1.83 ± 0.14 U mL–1, respectively). Expression of a mutant Erwinase proteoform – V293M – was also achieved and it presented a similar enzymatic activity. No significant loss in L-ASNase enzymatic activity was noticed after removal of cyclic AMP, spermidine, transfer RNA, T7 RNA polymerase and, especially, ammonium acetate (a common interference in ASNase enzymatic assays) from the CFPS reaction. CONCLUSION The protocol developed in this work will facilitate the screening of novel clinically-relevant L-ASNase proteoforms. © 2021 Society of Chemical Industry (SCI).

Perfil do doador de sangue autoexcluído no Hemocentro Regional de Uberaba-MG (HRU) no período de 1996 a 2006 / Self-exclusion profiles of blood donors of the Regional Blood Bank in Uberaba, Brazil (HRU) in the period of 1996 to 2006

Candidatos a doação são submetidos a triagem clínica e sorológica para minimizar o risco de transmissão de doenças via transfusão. Uma de suas limitações é a janela imunológica, que possibilita a transfusão de sangue contaminado. O objetivo deste trabalho foi avaliar o índice de autoexclusão de acordo com idade, gênero, estado civil, cor e tipo de doação, as variações anuais de autoexcluídos e sua eficácia em evitar a transfusão de sangue contaminado. Os dados foram analisados através do teste qui-quadrado, odds ratio e regressão linear. De 1996 a 2006, o Hemocentro Regional de Uberaba (HRU) coletou 176.097 bolsas de sangue, das quais 2,72 por cento foram desprezadas por autoexclusão, com significativo predomínio de homens, maiores de 29 anos, solteiros, não brancos e primeira doação (p<0,0001). Observou-se associação entre fidelização e autoexclusão, sugerindo que maior fidelização contribui para menor autoexclusão. A sorologia positiva para HIV1 (0,35 por cento) e HIV2 (0,23 por cento) foi significativamente maior nos autoexcluídos (p<0,0001), significância não observada para HCV (0,52 por cento) (p=0,24). Nos não autoexcluídos, estes percentuais foram de 0,15 por cento, 0,03 por cento e 0,41 por cento, respectivamente. A maior frequência de autoexclusão em homens maiores de 29 anos, solteiros e não brancos está de acordo, em parte, com o perfil do doador do HRU. O decréscimo de 1996 a 2001 é explicado por fatores comportamentais como criação do Centro de Testagem Anônima e maior fidelização dos doadores ao longo dos anos. A maior frequência de positividade nos autoexcluídos e três soroconversões em doações subsequentes reforçam a importância dessa ferramenta na diminuição do risco de janela imunológica.

Blood donor candidates are submitted to clinical and serological screening to minimize the risk of disease transmission. One of the limitations of this screening is the immunological window, a period that may contribute to the transfusion of contaminated blood. The aim of this study was to evaluate self-exclusion rates related to age, gender, marital status, race, type of donation, annual variations in self-exclusion and the efficacy of self-exclusion to prevent the transfusion of contaminated blood. A retrospective study was conducted and the data were analyzed using the chi-square test, odds ratio and linear regression. Of the 176,097 blood bags collected at the Uberaba Regional Blood Center (HRU) between 1996 and 2006, 2.72 percent were discarded due to self-exclusion. There was a predominance of first-time, unmarried, non-white male donors over the age of 29 years old (p<0.0001). An inverse association was observed between long-term commitment and self-exclusion, suggesting that the higher the return rate, the lower the incidence of self-exclusion. Positive serology for HIV1 (0.35 percent) and HIV2 (0.23 percent) was significantly higher among self-exclusion donors (p<0.0001), a fact not observed for HCV (0.52 percent) (p=0.24). The percentages of these three diseases were 0.15 percent, 0.03 percent and 0.41 percent, respectively among donors not initiating self-exclusion. The higher frequency of self-exclusion among unmarried, non-white male donors over the age of 29 partly agrees with the donor profile of HRU. The decline observed from 1996 to 2001 might be explained by behavioral factors, such as the creation of anonymous testing centers and an increase in the long-term commitment of donors over the years. The higher frequency of positivity among self-exclusion donors and only three seroconversions in subsequent donations support the importance of this tool to reduce the risk of contamination due to the immunological window.