Industrial Agile Transformations Lacking Business Emphasis: Results from a Nordic Survey Study

Peer reviewe

A chromatographic network for the purification of detergent-solubilized six-transmembrane epithelial antigen of the prostate 1 from Komagataella pastoris mini-bioreactor lysates

Funding Information: The authors acknowledge the support from FEDER funds through the POCI-COMPETE 2020–Operational Programme Competitiveness and Internationalisation in Axis I–Strengthening Research, Technological Development and Innovation (Project POCI-01-0145-FEDER-007491), Jorge Barroca-Ferreira's and Ana M. Gonçalves's individual PhD Fellowships (SFRH/BD/130068/2017 and SFRH/BD/147519/2019, respectively), and Luís A. Passarinha's sabbatical fellowship (SFRH/BSAB/150376/2019) from FCT–Fundação para a Ciência e Tecnologia. This work was also supported by the Health Sciences Research Centre CICS-UBI (UIDB/00709/2020 and UIDP/00709/2020), the Applied Molecular Biosciences Unit UCIBIO (UIDB/04378/2020 and UIDP/04378/2020) and the Associate Laboratory Institute for Health and Bioeconomy–i4HB (project LA/P/0140/2020) which are financed by National Funds from FCT/MCTES. Publisher Copyright: © 2022The Six-Transmembrane Epithelial Antigen of the Prostate 1 (STEAP1) is an integral membrane protein involved in cellular communications, in the stimulation of cell proliferation by increasing Reactive Oxygen Species levels, and in the transmembrane-electron transport and reduction of extracellular metal-ion complexes. The STEAP1 is particularly over-expressed in prostate cancer, in contrast with non-tumoral tissues and vital organs, contributing to tumor progression and aggressiveness. However, the current understanding of STEAP1 lacks experimental data on the respective molecular mechanisms, structural determinants, and chemical modifications. This scenario highlights the relevance of exploring the biosynthesis of STEAP1 and its purification for further bio-interaction and structural characterization studies. In this work, recombinant hexahistidine-tagged human STEAP1 (rhSTEAP1-His6) was expressed in Komagataella pastoris (K. pastoris) mini-bioreactor methanol-induced cultures and successfully solubilized with Nonidet P-40 (NP-40) and n-Decyl-β-D-Maltopyranoside (DM) detergents. The fraction capacity of Phenyl-, Butyl-, and Octyl-Sepharose hydrophobic matrices were evaluated by manipulating the ionic strength of binding and elution steps. Alternatively, immobilized metal affinity chromatography packed with nickel or cobalt were also studied in the isolation of rhSTEAP1-His6 from lysate extracts. Overall, the Phenyl-Sepharose and Nickel-based resins provided the desired selectivity for rhSTEAP1-His6 capture from NP-40 and DM detergent-solubilized K. pastoris extracts, respectively. After a polishing step using the anion-exchanger Q-Sepharose, a highly pure, fully solubilized, and immunoreactive 35 kDa rhSTEAP1-His6 fraction was obtained. Altogether, the established reproducible strategy for the purification of rhSTEAP1-His6 paves the way to gather additional insights on structural, thermal, and environmental stability characterization significantly contributing for the elucidation of the functional role and oncogenic behavior of the STEAP1 in prostate cancer microenvironment.publishersversionpublishe

Hemorheologic profile in patients with angina pectoris

Thirteen patients (9 men and 4 women) with stable angina, whose ages ranged from 48 to 62 years (mean 57 years), were studied in order to establish the hemorheologic profile consequent to effort as well as some parameters of the oxygen transport system. The results were compared with those obtained from a control group of 8 healthy individuals, 4 men and 4 women, whose ages ranged from 40 to 57 years (mean 48 years). The patients presented values of P50 in vivo during rest (R) higher than the control group (p < 0.05); the filtration time of blood (FT) and the flow rate of erythrocyte filtration (FR) rose significantly from the R to the precocious recovery (Rec) phase, both in the control group (p < 0.01) and in the patients (p < 0.001). There was, therefore, no remarkable difference in these parameters as well as in arterial pH, Hb and Ht at rest and after maximum effort, between the patients and the control group. The lactate presented higher values (p < 0.05) in the control group when only early phase of Rec was taken into account. In the patients there was a significant positive correlation during rest between 2,3-DPG and FT (p < 0.05) and between PSOiv and FT (p < 0.02). This correlation was not found in the control group. It was also found a significant rise of Hi in the early phase of Rec among the patients as opposed to the control group. The results suggest: (a) adaptation of the oxygen transport system in the patients with angina through a rise of oxyhemoglobin dissociation capacity; (b) aggravation of hemorheologic profile after effort, more conspicuous in the patients than in the control group; (c) the system of oxygen transport in the patients seems to be influenced by alterations of hernoiheologic parameters

Suitability of PLLA as piezoelectric substrates for tissue engineering evidenced by microscopy techniques

Since the discovery of the piezoelectric character of bone, the suitability of some piezoelectric materials have been studied for bone repair; they are thought to act like transducers converting the mechanical energy of skeletal deformation in electrical stimuli capable of controlling osteogenic growth. The mechanisms underlying this process are far from being understood and systematic studies at a local scale are required. Atomic force microscopy (AFM) is a unique way to observe phenomena at the nanoscale and liquid imaging provides a unique tool to assess biological phenomena at the nanoscale. So in this study, aiming at a better understanding of the role of piezoelectricity in the osteogenic growth, the interaction between a poled piezoelectric material, in this case poly (L-lactic) acid and an adhesion promoting protein, the fibronectin, and bone-like cells is evaluated by scanning probe microscopy and confocal laser scanning microscopy (CLSM). © Microscopy Society of America 2012.(undefined

Multicaloric effect in a multiferroic composite of Gd-5(Si,Ge)(4) microparticles embedded into a ferroelectric PVDF matrix

CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOThe coupling between electric, magnetic and elastic features in multiferroic materials is an emerging field in materials science, with important applications on alternative solid-state cooling technologies, energy harvesting and sensors/actuators. In this direction, we developed a thorough investigation of a multiferroic composite, comprising magnetocaloric/magnetostrictive Gd5Si2.4Ge1.6 microparticles blended into a piezo- and pyroelectric poly(vinylidene) fluoride (PVDF) matrix. Using a simple solvent casting technique, the formation and stabilization of PVDF electroactive phases are improved when the filler content increases from 2 to 12 weight fraction (wt.%). This effect greatly contributes to the magnetoelectric (ME) coupling, with the ME coefficient alpha(ME) increasing from 0.3 V/cm.Oe to 2.2 V/cm.Oe, by increasing the amount of magnetic material. In addition, magnetic measurements revealed that the ME-coupling has influenced the magnetocaloric effect via a contribution from the electroactive polymer and hence leading to a multicaloric effect. These results contribute to the development of multifunctional systems for novel technologies.9CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO203180/2014-3This work is funded by FEDER funds through the COMPETE 2020 Programme and National Funds throught FCT -Portuguese Foundation for Science and Technology under the projects POCI-01-0145-FEDER-029454, POCI-01-0145-FEDER-032527 and UID/FIS/04564/2016. This work was also supported by NECL with the project NORTE-01-0145-FEDER-022096 and by the European Union Horizon -2020 research and innovation program under the Marie Sklodowska-Curie Grant Agreement No. 734801. The authors acknowledge K. Pirota and F. Beron for the FTIR measurements performed at Unicamp in Brazil. Special thanks to Dr. A. Aliev for help in automatizing the ME experiments at Amirkhanov Institute of Physics and the helpful discussions. Federal Fluminense University, Brazil, permanent address for MSR; Aveiro University, Portugal, temporary address during this work for MSR. VMA thanks the CNPq for the Grant No. 203180/2014-3. A.A. acknowledges Russian Science Foundation for support magnetoelectric studies (grant No. 18-79-10176). J.H. Belo thanks FCT for Grant No. SFRH/BD/88440/2012, Project PTDC/FISMA/31302/2017, and his contract No. DL57/2016 reference SFRH-BPD-87430/2012. Finally, the discussion of results has been made possible through the mobility grant provided by the 5 top 100 Russian Academic Excellence Project at the Immanuel Kant Baltic Federal University

Control of developmentally primed erythroid genes by combinatorial co-repressor actions

How transcription factors (TFs) cooperate within large protein complexes to allow rapid modulation of gene expression during development is still largely unknown. Here we show that the key haematopoietic LIM-domain-binding protein-1 (LDB1) TF complex contains several activator and repressor components that together maintain an erythroid-specific gene expression programme primed for rapid activation until differentiation is induced. A combination of proteomics, functional genomics and in vivo studies presented here identifies known and novel co-repressors, most notably the ETO2 and IRF2BP2 proteins, involved in maintaining this primed state. The ETO2-IRF2BP2 axis, interacting with the NCOR1/SMRT co-repressor complex, suppresses the expression of the vast majority of archetypical erythroid genes and pathways until its decommissioning at the onset of terminal erythroid differentiation. Our experiments demonstrate that multimeric regulatory complexes feature a dynamic interplay between activating and repressing components that determines lineage-specific gene expression and cellular differentiation

Pregnancy in the mature adult mouse does not alter the proportion of mammary epithelial stem/progenitor cells

Introduction In humans, an early full-term pregnancy reduces lifetime breast cancer risk by up to 50% whereas a later pregnancy (>35 years old) can increase lifetime risk. Several mechanisms have been suggested, including changes in levels of circulating hormones, changes in the way the breast responds to these hormones, changes in gene expression programmes which may alter susceptibility to transformation and changes to mammary stem cell numbers or behaviour. Previous studies have shown that the mammary tissue isolated from both virgin and parous mice has the ability to repopulate a cleared mammary fat pad in transplant experiments. Limited dilution transplant assays have demonstrated that early pregnancy (at 5 weeks of age) reduces stem/progenitor cell numbers in the mouse mammary epithelium by twofold. However, the effects on stem/progenitor cell numbers in the mammary epithelium of a pregnancy in older animals have not yet been tested. Methods Mice were put through a full-term pregnancy at 9 weeks of age, when the mammary epithelium is mature. The total mammary epithelium was purified from parous 7-week post-lactation and age-matched virgin mice and analysed by flow cytometry and limiting dilution cleared fat pad transplants. Results There were no significant differences in the proportions of different mammary epithelial cell populations or numbers of CD24+/Low Sca-1- CD49fHigh cells (stem cell enriched basal mammary epithelial compartment). There was no significant difference in stem/progenitor cell frequency based on limiting dilution transplants between the parous and age-matched virgin epithelium. Conclusions Although differences between parous and virgin mammary epithelium at later time points post lactation or following multiple pregnancies cannot be ruled out, there are no differences in stem/progenitor cell numbers between mammary epithelium isolated from parous animals which were mated at 9 weeks old and virgin animals. However, a recent report has suggested that animals that were mated at 5 weeks old have a twofold reduction in stem/progenitor cell numbers. This is of interest given the association between early, but not late, pregnancy and breast cancer risk reduction in humans. However, a mechanistic connection between stem cell numbers and breast cancer risk remains to be established