human embryonic kidney cells
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2022 ◽  
Vol 15 (1) ◽  
pp. 71
Author(s):  
Enas Al-Ani ◽  
Wayne Heaselgrave

The aim of this study was to investigate the potential of thymol to inhibit Candida biofilm formation and improve thymol biocompatibility in the presence of hydroxypropyl methylcellulose (HPMC) and poloxamer 407 (P407), as possible drug carriers. Thymol with and without polymers were tested for its ability to inhibit biofilm formation, its effect on the viability of biofilm and biocompatibility studies were performed on HEK 293 (human embryonic kidney) cells. Thymol showed a concentration dependent biofilm inhibition; this effect was slightly improved when it was combined with HPMC. The Thymol-P407 combination completely inhibited the formation of biofilm and the antibiofilm effect of thymol decreased as the maturation of Candida biofilms increased. The effect of thymol on HEK 293 cells was a loss of nearly 100% in their viability at a concentration of 250 mg/L. However, in the presence of P407, the viability was 25% and 85% using neutral red uptake and sulforhodamine B assays, respectively. While, HPMC had less effect on thymol activity the thymol-P407 combination showed a superior inhibitory effect on biofilm formation and better biocompatibility with human cell lines. The combination demonstrates a potential medical use for the prevention of Candida biofilm formation.


2022 ◽  
Vol 17 (1) ◽  
pp. 1934578X2110689
Author(s):  
Ari S. Nugraha ◽  
Yoshinta D. Purnomo ◽  
Antonius N. Widhi Pratama ◽  
Bawon Triatmoko ◽  
Rudi Hendra ◽  
...  

Malaria is a neglected tropical disease that still demands serious efforts to tackle successfully, including the need for new antimalarial lead compounds to combat drug-resistant Plasmodium. Intensive phytochemical and pharmacological investigation into the Indonesian medicinal plants Swietenia mahagoni and Pluchea indica successfully revealed 5 constituents. Antimalarial bioassays indicated 34,5-tri- O-caffeoylquinic acid (4) to be the most active against Plasmodium falciparum 3D7 and Dd2 strains with IC50 values of 8.2 and 8.8 µM, respectively. No cytotoxicity was observed against Human Embryonic Kidney cells at a concentration of 40 µM.


2021 ◽  
Author(s):  
Yu Shi ◽  
Shankar Sivarajan ◽  
Katherine M Xiang ◽  
Geran M Kostecki ◽  
Leslie Tung ◽  
...  

Abstract The actomyosin cytoskeleton enables cells to resist deformation, crawl, change their shape and sense their surroundings. Despite decades of study, how its molecular constituents can assemble together to form a network with the observed mechanics of cells remains poorly understood. Recently, it has been shown that the actomyosin cortex of quiescent cells can undergo frequent, abrupt reconfigurations and displacements, called cytoquakes. Notably, such fluctuations are not predicted by current physical models of actomyosin networks, and their prevalence across cell types and mechanical environments has not previously been studied. Using micropost array detectors, we have performed high-resolution measurements of the dynamic mechanical fluctuations of cells’ actomyosin cortex and stress fiber networks. This reveals cortical dynamics dominated by cytoquakes—intermittent events with a fat-tailed distribution of displacements, sometimes spanning microposts separated by 4 μm, in all cell types studied. These included 3T3 fibroblasts, where cytoquakes persisted over substrate stiffnesses spanning the tissue-relevant range of 4.3 kPa–17 kPa, and primary neonatal rat cardiac fibroblasts and myofibroblasts, human embryonic kidney cells and human bone osteosarcoma epithelial (U2OS) cells, where cytoquakes were observed on substrates in the same stiffness range. Overall, these findings suggest that the cortex self-organizes into a marginally stable mechanical state whose physics may contribute to cell mechanical properties, active behavior and mechanosensing.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Gunsmaa Nyamsuren ◽  
Gregor Rapp ◽  
Hassan Dihazi ◽  
Elisabeth M. Zeisberg ◽  
Desiree Tampe ◽  
...  

AbstractAryl hydrocarbon receptor nuclear translocator (ARNT) mediates anti-fibrotic activity in kidney and liver through induction of ALK3-receptor expression and subsequently increased Smad1/5/8 signaling. While expression of ARNT can be pharmacologically induced by sub-immunosuppressive doses of FK506 or by GPI1046, its anti-fibrotic activity is only realized when ARNT-ARNT homodimers form, as opposed to formation of ARNT-AHR or ARNT-HIF1α heterodimers. Mechanisms underlying ARNTs dimerization decision to specifically form ARNT–ARNT homodimers and possible cues to specifically induce ARNT homodimerization have been previously unknown. Here, we demonstrate that phosphorylation of the Ser77 residue is critical for ARNT–ARNT homodimer formation and stabilization. We further demonstrate that inhibition of PP2A phosphatase activity by LB100 enhances ARNT–ARNT homodimers both in vivo and in vitro (mouse tubular epithelial cells and human embryonic kidney cells). In murine models of kidney fibrosis, and also of liver fibrosis, combinations of FK506 or GPI1046 (to induce ARNT expression) with LB100 (to enhance ARNT homodimerization) elicit additive anti-fibrotic activities. Our study provides additional evidence for the anti-fibrotic activity of ARNT–ARNT homodimers and reveals Ser77 phosphorylation as a novel pharmacological target to realize the therapeutic potential of increased ARNT transactivation activity.


2021 ◽  
pp. 101549
Author(s):  
David A. Hanna ◽  
Courtney M. Moore ◽  
Liu Liu ◽  
Xiaojing Yuan ◽  
Iramofu M. Dominic ◽  
...  

AAPS Open ◽  
2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Sara Javadi ◽  
Negar Motakef Kazemi ◽  
Raheleh Halabian

AbstractThe oil-in-water (O/W) nano-emulsion (NE) is expanded to enhance the bioavailability of hydrophobic compounds. The NE can be prepared by herbal extract and essential oil as herbal medicines for antidiabetic treatment. In the present study, the O/W NE was prepared by fenugreek extract (FE), nettle extract (NE), and cumin essential oil (CEO) using tween 80 and span 80 surfactants in an ultrasonic bath, at room temperature within 18 min. The antidiabetic property was evaluated by determining glucose absorption using cultured rat L6 myoblast cell line (L6) myotubes and insulin secretion using the cultured mouse pancreatic beta-cell (RIN-5) for NEs. The samples were investigated by dynamic light scattering (DLS) to examine the size distribution and size, zeta potential for the charge determination, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) to investigate morphology and size. The rheological properties were studied by viscosity. The sample stability was evaluated at different temperatures and days by DLS and SEM analyses. The cytotoxicity of samples was explored by MTT assay for HEK293 human cell line as a specific cell line originally derived from human embryonic kidney cells at three different concentrations for three periods of time. The NEs with nanometer-size were observed with antidiabetic properties, low cytotoxicity, and suitable stability. This study provides definitive evidence for the NE as a plant medicine with antidiabetic properties. The NE can be a good candidate for biomedical applications.


2021 ◽  
Author(s):  
Tunglin Tsau ◽  
Prema Vyas ◽  
Lexi Crowell ◽  
Mary Tran ◽  
Destiney Ward ◽  
...  

Human mesenchymal stem cells (hMSCs) have gained traction in transplantation therapy due to their immunomodulatory, paracrine, immune-evasive, and multipotent differentiation potential. Given the heterogeneous nature of hMSCs, therapeutic treatments and robust in vivo and in vitro experiments require additional biomarkers to ensure reproducibility when using these stem cells. In this work, we utilized dielectrophoresis (DEP), a label-free electrokinetic phenomenon, to investigate and quantify the heterogeneity of hMSCs derived from the bone marrow (BM) and adipose tissue (AD). Through computer simulation, we identified that the transient slope of the DEP force spectra can be used as a metric of heterogeneity. The electrical properties of BM-hMSCs were compared to homogeneous mouse fibroblasts (NIH-3T3), human fibroblasts (WS1), and human embryonic kidney cells (HEK-293). BM-hMSCs DEP profile was most different from HEK-293 cells. We compared the DEP profiles of BM-hMSCs and AD-hMSCs and found they have similar membrane capacitances, differing cytoplasm conductivity, and transient slopes. Inducing both populations to differentiate into adipocyte and osteocyte cells revealed they behave differently in response to differentiation-inducing cytokines. Histology and RT-qPCR analyses of the differentiation-related genes revealed differences in heterogeneity between BM-hMSCs and AD-hMSCs. The differentiation profiles correlate well with the DEP profiles developed and indicate that these BM-hMSCs have higher differentiation potential than AD-hMSCs. Our results demonstrate using DEP, membrane capacitance, cytoplasm conductivity, and transient slope can uniquely characterize the inherent heterogeneity of hMSCs to guide robust and reproducible stem cell transplantation therapies.


Livers ◽  
2021 ◽  
Vol 1 (4) ◽  
pp. 221-229
Author(s):  
Melissa J. Ruggiero ◽  
Haley Miller ◽  
Jessica Y. Idowu ◽  
Jeremiah D. Zitzow ◽  
Shu-Ching Chang ◽  
...  

Na+/taurocholate cotransporting polypeptide (NTCP) is important for the enterohepatic circulation of bile acids, which has been suggested to contribute to the long serum elimination half-lives of perfluoroalkyl substances in humans. We demonstrated that some perfluoroalkyl sulfonates are transported by NTCP; however, little was known about carboxylates. The purpose of this study was to determine if perfluoroalkyl carboxylates would interact with NTCP and potentially act as substrates. Sodium-dependent transport of [3H]-taurocholate was measured in human embryonic kidney cells (HEK293) stably expressing NTCP in the absence or presence of perfluoroalkyl carboxylates with varying chain lengths. PFCAs with 8 (PFOA), 9 (PFNA), and 10 (PFDA) carbons were the strongest inhibitors. Inhibition kinetics demonstrated competitive inhibition and indicated that PFNA was the strongest inhibitor followed by PFDA and PFOA. All three compounds are transported by NTCP, and kinetics experiments revealed that PFOA had the highest affinity for NTCP with a Km value of 1.8 ± 0.4 mM. The Km value PFNA was estimated to be 5.3 ± 3.5 mM and the value for PFDA could not be determined due to limited solubility. In conclusion, our results suggest that, in addition to sulfonates, perfluorinated carboxylates are substrates of NTCP and have the potential to interact with NTCP-mediated transport.


Biomolecules ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1448
Author(s):  
Emiko Kinoshita-Kikuta ◽  
Eiji Kinoshita ◽  
Misaki Suga ◽  
Mana Higashida ◽  
Yuka Yamane ◽  
...  

The production of heterologous proteins is an important procedure for biologists in basic and applied sciences. A variety of cell-based and cell-free protein expression systems are available to achieve this. The expression system must be selected carefully, especially for target proteins that require post-translational modifications. In this study, human Src family kinases were prepared using six different protein expression systems: 293 human embryonic kidney cells, Escherichia coli, and cell-free expression systems derived from rabbit reticulocytes, wheat germ, insect cells, or Escherichia coli. The phosphorylation status of each kinase was analyzed by Phos-tag SDS-PAGE. The kinase activities were also investigated. In the eukaryotic systems, multiple phosphorylated forms of the expressed kinases were observed. In the rabbit reticulocyte lysate system and 293 cells, differences in phosphorylation status between the wild-type and kinase-dead mutants were observed. Whether the expressed kinase was active depended on the properties of both the kinase and each expression system. In the prokaryotic systems, Src and Hck were expressed in autophosphorylated active forms. Clear differences in post-translational phosphorylation among the protein expression systems were revealed. These results provide useful information for preparing functional proteins regulated by phosphorylation.


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