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Tissue engineered hyaline cartilage is plagued by poor mechanical properties largely due to inadequate type II collagen expression. Of note, commonly used defined chondrogenic media lack 14 vitamins and minerals, some of which are implicated in chondrogenesis. Type II collagen promoter-driven Gaussia luciferase was transfected into ATDC5 cells to create a chondrogenic cell with a secreted-reporter. UCL-TRO-1938 The reporter cells were used in an aggregate-based chondrogenic culture model to develop a high-throughput analytic platform. This high-throughput platform was used to assess the effect of vitamins and minerals, alone and in combination with TGFβ1, on COL2A1 promoter-driven expression. Significant combinatorial effects between vitamins, minerals, and TGFβ1 in terms of COL2A1 promoter-driven expression and metabolism were discovered. UCL-TRO-1938 An "optimal" continual supplement of copper and vitamin K in the presence of TGFβ1 gave a 2.5-fold increase in COL2A1 promoter-driven expression over TGFβ1 supplemented media alone in ATDC5 cells. Copyright © 2020 Dennis, Splawn and Kean.With the continued steep rise of the global human population, and the paucity of safe and practical contraceptive options available to men, the need for development of effective and reversible non-hormonal methods of male fertility control is widely recognized. UCL-TRO-1938 Currently there are several contraceptive options available to men, however, none of the non-hormonal alternatives have been clinically approved. To advance progress in the development of a safe and reversible contraceptive for men, further identification of novel reproductive tract-specific druggable protein targets is required. Here we provide an overview of genes/proteins identified in the last decade as specific or highly expressed in the male reproductive tract, with deletion phenotypes leading to complete male infertility in mice. link2 These phenotypes include arrest of spermatogenesis and/or spermiogenesis, abnormal spermiation, abnormal spermatid morphology, abnormal sperm motility, azoospermia, globozoospermia, asthenozoospermia, and/or teratozoospermia, which are all desirable outcomes for a novel male contraceptive. We also consider other associated deletion phenotypes that could impact the desirability of a potential contraceptive. We further discuss novel contraceptive targets underscoring promising leads with the objective of presenting data for potential druggability and whether collateral effects may exist from paralogs with close sequence similarity. link2 Copyright © 2020 Kent, Johnston, Strump and Garcia.Successful mitotic cell division is critically dependent on the formation of correct attachments between chromosomes and spindle microtubules. Microtubule attachments are mediated by kinetochores, which are large proteinaceous structures assembled on centromeric chromatin of mitotic chromosomes. These attachments must be sufficiently stable to transduce force; however, the strength of these attachments are also tightly regulated to ensure timely, error-free progression through mitosis. The highly conserved, kinetochore-associated NDC80 complex is a core component of the kinetochore-microtubule attachment machinery in eukaryotic cells. A small, disordered region within the Hec1 subunit of the NDC80 complex - the N-terminal "tail" domain - has been actively investigated during the last decade due to its roles in generating and regulating kinetochore-microtubule attachments. In this review, we discuss the role of the NDC80 complex, and specifically the Hec1 tail domain, at the kinetochore-microtubule interface, and how recent studies provide a more unified view of Hec1 tail domain function. link2 Copyright © 2020 Wimbish and DeLuca.For many years now, Bacillus megaterium serves as a microbial workhorse for the high-level production of recombinant proteins in the g/L-scale. However, efficient and stable production processes require the knowledge of the molecular adaptation strategies of the host organism to establish optimal environmental conditions. Here, we interrogated the osmotic stress response of B. megaterium using transcriptome, proteome, metabolome, and fluxome analyses. An initial transient adaptation consisted of potassium import and glutamate counterion synthesis. The massive synthesis of the compatible solute proline constituted the second longterm adaptation process. Several stress response enzymes involved in iron scavenging and reactive oxygen species (ROS) fighting proteins showed higher levels under prolonged osmotic stress induced by 1.8 M NaCl. At the same time, the downregulation of the expression of genes of the upper part of glycolysis resulted in the activation of the pentose phosphate pathway (PPP), generating an oversupply of NADPH. The increased production of lactate accompanied by the reduction of acetate secretion partially compensate for the unbalanced (NADH/NAD+) ratio. Besides, the tricarboxylic acid cycle (TCA) mainly supplies the produced NADH, as indicated by the higher mRNA and protein levels of involved enzymes, and further confirmed by 13C flux analyses. As a consequence of the metabolic flux toward acetyl-CoA and the generation of an excess of NADPH, B. megaterium redirected the produced acetyl-CoA toward the polyhydroxybutyrate (PHB) biosynthetic pathway accumulating around 30% of the cell dry weight (CDW) as PHB. This direct relation between osmotic stress and intracellular PHB content has been evidenced for the first time, thus opening new avenues for synthesizing this valuable biopolymer using varying salt concentrations under non-limiting nutrient conditions. link3 Copyright © 2020 Godard, Zühlke, Richter, Wall, Rohde, Riedel, Poblete-Castro, Krull and Biedendieck.DNA-functionalized gold nanoparticles (AuNPs) have been widely used in directed assembly of materials, biosensors, and drug delivery. This conjugate may encounter proteins in these applications and proteins may affect not only DNA adsorption but also the function of the attached DNA. Bovine serum albumin (BSA) with many cysteine residues can strongly adsorb on AuNPs and this conjugate showed high colloidal stability against salt, acid and base. Similar protection effects were also observed with a few other common proteins including catalase, hemoglobin, glucose oxidase, and horseradish peroxidase. DNA oligonucleotides without a thiol label can hardly displace adsorbed BSA, and BSA cannot displace pre-adsorbed DNA either, indicating a strongly kinetically controlled system. Thiolated DNA can be attached at a low density on the AuNPs with a BSA corona. link3 The BSA corona did not facilitate the hybridization of the conjugated DNA, while a smaller peptide, glutathione allowed faster hybridization. Overall, proteins increase the colloidal stability of AuNPs, and they do not perturb the gold-thiol bond in the DNA conjugate, although a large protein corona may inhibit the hybridization function of DNA. Copyright © 2020 Wu, Peng, Zhu, Jiang and Liu.The role of conductive carbon additive on the electrode/electrolyte interface formation mechanism was examined in the low-potential (3.0-0 V) and high-potential (3.0-4.7 V) regions. Here the most commonly used conductive carbon Super P was used to prepared electrode with polyvinylidene fluoride binder without any active material. link3 The dynamic process of interface formation was observed with in situ Scanning Electrochemical Microscopy. The electronically insulating electrode/electrolyte passivation layer with areal heterogeneity was formed after cycles in both potential regions. The low-potential interface layer is mainly composed of inorganic compounds covering the conductive carbon surface; While the electrode after high-potential sweep tends to lose its original carbon structure and has more organic species formed on its surface. Copyright © 2020 Liu, Zeng, Liu, Wang, Zhang, Zhao, Kang and Li.Natural protein-based hydrogels possess excellent biocompatibility; however, most of them are weak or brittle. In the present work, high strength hybrid dual-crosslinking BSA gels (BSA DC gels), which have both chemical cross-linking and physical cross-linking, were fabricated by a facile photoreaction-heating process. BSA DC gels showed high transparency (light transmittance of ~90%) and high strength. At optimal conditions, BSA DC gel exhibited high compressive strength (σc,f) of 37.81 ± 2.61 MPa and tensile strength (σt,f) of 0.62 ± 0.078 MPa, showing it to be much stronger than physically cross-linked BSA gel (BSA PC gel) and chemically cross-linked BSA gel (BSA CC gel). More importantly, BSA DC gel displayed non-swelling properties while maintaining high strength in DI water, pH = 3.0, and pH = 10.0. Moreover, BSA DC gel also demonstrated large hysteresis, rapid self-recovery, and excellent fatigue resistance properties. It is believed that our BSA DC gel can potentially be applied in biomedical fields. Copyright © 2020 Lu, Zhu, Wang, Liu, Tang, Sun, Yang, Qin, Sun and Chen.In this study, micro/nanoscale LiFePO4/graphene composites are synthesized successfully using a one-step microwave heating method. One-step microwave heating can simplify the reduction step of graphene oxide and provide a convenient, economical, and effective method of preparing graphene composites. The structural analysis shows that LiFePO4/graphene has high phase purity and crystallinity. The morphological analysis shows that LiFePO4/graphene microspheres and micron blocks are composed of densely aggregated nanoparticles; the nanoparticle size can shorten the diffusion path of lithium ions and thus increase the lithium-ion diffusion rate. Additionally, the graphene sheets can provide a rapid transport path for electrons, thus increasing the electronic conductivity of the material. Furthermore, the nanoparticles being packed into the micron graphene sheets can ensure stability in the electrolyte during charging and discharging. Raman analysis reveals that the graphene has a high degree of graphitization. Electrochemical analysis shows that the LiFePO4/graphene has an excellent capacity, high rate performance, and cycle stability. The discharge capacities are 166.3, 156.1, 143.0, 132.4, and 120.9 mAh g-1 at rates of 0.1, 1, 3, 5, and 10 C, respectively. The superior electrochemical performance can be ascribed to the synergy of the shorter lithium-ion diffusion path achieved by LiFePO4 nanoparticles and the conductive networks of graphene. Copyright © 2020 Liu, Yan, Li, Zhang and Sun.Both vertebrates and invertebrates display active innate immune mechanisms for defense against microbial infection, including diversified repertoires of soluble and cell-associated lectins that can effect recognition and binding to potential pathogens, and trigger downstream effector pathways that clear them from the host internal milieu. Galectins are widely distributed and highly conserved lectins that have key regulatory effects on both innate and adaptive immune responses. In addition, galectins can bind to exogenous ("non-self") carbohydrates on the surface of bacteria, enveloped viruses, parasites, and fungi, and function as recognition receptors and effector factors in innate immunity. Like most invertebrates, eastern oysters (Crassostrea virginica) and softshell clams (Mya arenaria) can effectively respond to most immune challenges through soluble and hemocyte-associated lectins. The protozoan parasite Perkinsus marinus, however, can infect eastern oysters and cause "Dermo" disease, which is highly detrimental to both natural and farmed oyster populations.
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