Towards a generic prototyping approach for therapeutically-relevant peptides and proteins in a cell-free translation system

Advances in peptide and protein therapeutics increased the need for rapid and cost-effective polypeptide prototyping. While in vitro translation systems are well suited for fast and multiplexed polypeptide prototyping, they suffer from misfolding, aggregation and disulfide-bond scrambling of the translated products. Here we propose that efficient folding of in vitro produced disulfide-rich peptides and proteins can be achieved if performed in an aggregation-free and thermodynamically controlled folding environment. To this end, we modify an E. coli-based in vitro translation system to allow co-translational capture of translated products by affinity matrix. This process reduces protein aggregation and enables productive oxidative folding and recycling of misfolded states under thermodynamic control. In this study we show that the developed approach is likely to be generally applicable for prototyping of a wide variety of disulfide-constrained peptides, macrocyclic peptides with non-native bonds and antibody fragments in amounts sufficient for interaction analysis and biological activity assessment.

Hybrid Biochar/Ceria Nanomaterials: Synthesis, Characterization and Activity Assessment for the Persulfate-Induced Degradation of Antibiotic Sulfamethoxazole

Biochar from spent malt rootlets was employed as the template to synthesize hybrid biochar-ceria materials through a wet impregnation method. The materials were tested for the activation of persulfate (SPS) and subsequent degradation of sulfamethoxazole (SMX), a representative antibiotic, in various matrices. Different calcination temperatures in the range 300–500 °C were employed and the resulting materials were characterized by means of N2 adsorption and potentiometric mass titration as well as TGA, XRD, SEM, FTIR, DRS, and Raman spectroscopy. Calcination temperature affects the biochar content and the physicochemical properties of the hybrid materials, which were tested for the degradation of 500 μg L−1 SMX with SPS (in the range 200–500 mg L−1) in various matrices including ultrapure water (UPW), bottled water, wastewater, and UPW spiked with bicarbonate, chloride, or humic acid. Materials calcined at 300–350 °C, with a surface area of ca. 120 m2 g−1, were the most active, yielding ca. 65% SMX degradation after 120 min of reaction in UPW; materials calcined at higher temperatures as well as bare biochar were less active. Degradation decreased with increasing matrix complexity due to the interactions amongst the surface, the contaminant, and the oxidant. Experiments in the presence of scavengers (i.e., methanol, t-butanol, and sodium azide) revealed that sulfate and hydroxyl radicals as well as singlet oxygen were the main oxidative species.

AuNP Aptasensor for Hodgkin Lymphoma Monitoring

A liquid biopsy based on circulating small extracellular vesicles (SEVs) has not yet been used in routine clinical practice due to the lack of reliable analytic technologies. Recent studies have demonstrated the great diagnostic potential of nanozyme-based systems for the detection of SEV markers. Here, we hypothesize that CD30-positive Hodgkin and Reed–Sternberg (HRS) cells secrete CD30 + SEVs; therefore, the relative amount of circulating CD30 + SEVs might reflect classical forms of Hodgkin lymphoma (cHL) activity and can be measured by using a nanozyme-based technique. A AuNP aptasensor analytics system was created using aurum nanoparticles (AuNPs) with peroxidase activity. Sensing was mediated by competing properties of DNA aptamers to attach onto surface of AuNPs inhibiting their enzymatic activity and to bind specific markers on SEVs surface. An enzymatic activity of AuNPs was evaluated through the color reaction. The study included characterization of the components of the analytic system and its functionality using transmission and scanning electron microscopy, nanoparticle tracking analysis (NTA), dynamic light scattering (DLS), and spectrophotometry. AuNP aptasensor analytics were optimized to quantify plasma CD30 + SEVs. The developed method allowed us to differentiate healthy donors and cHL patients. The results of the CD30 + SEV quantification in the plasma of cHL patients were compared with the results of disease activity assessment by positron emission tomography/computed tomography (PET-CT) scanning, revealing a strong positive correlation. Moreover, two cycles of chemotherapy resulted in a statistically significant decrease in CD30 + SEVs in the plasma of cHL patients. The proposed AuNP aptasensor system presents a promising new approach for monitoring cHL patients and can be modified for the diagnostic testing of other diseases.

PENINGKATAN PEMAHAMAN KONSEP SISWA DENGAN PENDEKATAN METAPHORICAL THINKING PADA PEMBELAJARAN DARING MATERI ARITMATIKA SOSIAL

This research is motivated by the low understanding of students' mathematical concepts, especially on social arithmetic material. The purpose of this study was to describe the stages of the metaphorical thinking approach method and the magnitude of the increase in students' conceptual understanding of social arithmetic material. The data collection method used in this classroom action research (CAR) includes written tests and observations in each cycle. The research instruments used were lesson plans, worksheets, question sheets, and observation sheets for the implementation of learning in each cycle. The research subjects were students of class VII in one of the madrasah tsanawiyah in Lamongan as many as 23 students. Based on the results of data analysis, the classical student activity assessment increased 23.85% from 66.25% to 90.1%. Student test results also increased by 34.8%, namely from cycle I it reached 47.8% and cycle II reached 82.6%, with the results achieved can be declared complete, and it can be concluded that in online mathematics learning there is an increase in understanding concept in class VII madrasah tsanawiyah in Lamongan by using the application of Metaphorical Thinking.

The Calcivis story - enamel caries activity assessment from technology to practice

The Calcivis story is one of innovation and collaboration to deliver new technology capable of helping dentists improve patient care through solving an unmet clinical need in assessing the activity of caries lesions in enamel. Presently, there is no system routinely used in dental practice that can, in a single visit, determine whether a non-cavitated caries lesion is active or not. Calcivis has evolved since 2005, when a potential link between basic science in luminescence and differentiating initial-stage caries lesions that are actively demineralising and likely to progress, from other lesions which are inactive and currently do not need interventive care, was recognised. The 16-year journey has involved clinical academic dentists, scientists and entrepreneurs, general practitioners and their patients, together with serial investors and a core team working to patent, refine, assess and develop products to submit to regulatory approval and take to the international dental market. This journey has been made possible through effective long-term collaborations between disparate groups all sharing a common vision for the possibilities of harnessing new technology to help dental professionals provide better care for their patients.