Human Embryonic Kidney Cell Line Research Articles

Quantum Dot Erythropoietin Bioconjugates Enhance EPO-Receptor Clustering on Transfected Human Embryonic Kidney Cells.

Erythropoietin (EPO)-induced cellular signaling through the EPO receptor (EPOR) is a fundamental pathway for the modulation of cellular behavior and activity. In our previous work, we showed in primary human astrocytes that the multivalent display of EPO on the surface of semiconductor quantum dots (QDs) mediates augmented JAK/STAT signaling, a concomitant 1.8-fold increase in the expression of aquaporin-4 (AQPN-4) channel proteins, and a 2-fold increase in the AQPN-4-mediated water transport activity. Our hypothesis is that this enhanced signaling involves the simultaneous ligation and clustering of EPOR by QD-EPO conjugates. Here, we utilized a human embryonic kidney (HEK 293T/17) cell line transfected with EPOR fused to enhanced green fluorescent protein (eGFP) to visualize EPOR clustering. We demonstrate that QDs displaying five copies of EPO (bearing a C-terminal 6-histidine tract) on the nanoparticle surface induce a 1.8-fold increase in EPOR clustering compared to monomeric EPO at the same concentration. Our findings confirm the critical role played by the multivalent display of EPO in mediating clustering of the EPOR. More generally, these results illustrate the capability of nanoparticle-growth factor bioconjugates to control the activity of cognate receptors and the important role played by multivalent display in the modulation of selective cellular delivery and signaling.

Multiplex genome editing eliminates lactate production without impacting growth rate in mammalian cells.

The Warburg effect, which describes the fermentation of glucose to lactate even in the presence of oxygen, is ubiquitous in proliferative mammalian cells, including cancer cells, but poses challenges for biopharmaceutical production as lactate accumulation inhibits cell growth and protein production. Previous efforts to eliminate lactate production in cells for bioprocessing have failed as lactate dehydrogenase is essential for cell growth. Here, we effectively eliminate lactate production in Chinese hamster ovary and in the human embryonic kidney cell line HEK293 by simultaneous knockout of lactate dehydrogenases and pyruvate dehydrogenase kinases, thereby removing a negative feedback loop that typically inhibits pyruvate conversion to acetyl-CoA. These cells, which we refer to as Warburg-null cells, maintain wild-type growth rates while producing negligible lactate, show a compensatory increase in oxygen consumption, near total reliance on oxidative metabolism, and higher cell densities in fed-batch cell culture. Warburg-null cells remain amenable for production of diverse biotherapeutic proteins, reaching industrially relevant titres and maintaining product glycosylation. The ability to eliminate lactate production may be useful for biotherapeutic production and provides a tool for investigating a common metabolic phenomenon.

Quinic acid enhances kanamycin efficacy against methicillin-resistant Staphylococcus aureus biofilms.

Methicillin-resistant Staphylococcus aureus (MRSA) form biofilms that contribute to increased antimicrobial resistance, leading to treatment failure and/or relapse. It is, therefore, necessary to develop new antibiofilm strategies to eradicate MRSA biofilms related infections. This study was aimed to evaluate the effect of the combination of quinic acid and kanamycin against the preformed biofilms of methicillin-resistant Staphylococcus aureus. Broth microdilution method was deployed to evaluate the antibacterial activity. Whereas antibiofilm activity was evaluated by crystal violet staining, 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium-bromide (MTT) assay, and scanning electron microscopy (SEM). The checkerboard method was adopted to assess the combination effects. Quantification of exopolysaccharides was determined by the phenol-sulfuric acid method. The eDNA was quantified by fluorescence spectrophotometry. Cytotoxicity activity was evaluated by the MTT assay on the human embryonic kidney (HEK 293) cell line. Quinic acid, combined with kanamycin, effectively eradicated the methicillin-resistant S. aureus biofilms by effecting biofilm biomass and cell viability. Scanning electron microscopy demonstrated a less adherence of S. aureus cells, - after treatment with quinic acid combined with kanamycin, as compared to each drug alone. The combination of quinic acid and kanamycin thus demonstrated the ability to destroy the exopolysaccharides and eDNA of biofilm matrix without any toxic effect on HEK 293cells. Our results demonstrated the potential of using quinic acid in combination therapy, with an antibiotic, against infections caused by MRSA strains.

MC-LR induced apoptosis in human embryonic kidney (HEK293) cells through activation of TNF-R1/RIPK1 pathway

ABSTRACT In recent years, the outbreak of cyanobacterial blooms has become increasingly frequent. Microcystin-LR (MC-LR), a metabolite of cyanobacteria, poses a significant threat to the ecosystem and human health. Several studies have demonstrated that MC-LR might induce renal cell apoptosis, as a consequence of tissue damage. However, the molecular mechanisms underlying MC-LR-initiated renal injury remain to be determined. This investigation aimed to determine the role of apoptosis in MC-LR-induced kidney damage and its potential underlying mechanisms using the human embryonic kidney (HEK293) cell line. The results of TUNEL and immunofluorescence assays indicated that MC-LR induced increased apoptosis in HEK293 cells. Compared to control, the mRNA expression levels of RIPK1, caspase-8, and TNF-α were elevated following incubation with MC-LR, while the mRNA expression level of Bcl-2/Bax was decreased. The protein levels of RIPK1, TNF-R1, and caspase-8 were elevated in the MC-LR-treated HEK293 cells. Data demonstrated that MC-LR induced renal cell apoptosis through activation of the TNF-R1/RIPK1 pathway, providing new insights into understanding the toxic mechanisms attributed to MC-LR.

Design and Synthesis of Propargyloxy Functionalized Pyrazole-Based Aurones: Exploring Their Potent Anticancer Properties Against AGS and MCF-7 Cell Lines.

FDA-approved numerous commercial and natural drugs used in cancer treatment feature either pyrazole or alkyne moieties. On the basis of this, we designed and synthesized 20 novel propargyloxy-substituted pyrazole-based aurones (10a-j and 11a-j) and evaluated for their anticancer potential against cancerous MCF-7 and human gastric adenocarcinoma (AGS)cell lines, as well as normal cell line human embryonic kidney 293 (HEK-293), through MTT assay. Among these tested compounds, five (10d-f, 11e, and 11f) displayed potent cytotoxic properties for AGS cancer cell line with IC50 values ranging from 19.7 to 28.5µM, better than the reference drugs leucovorin (IC50=30.8µM) and oxaliplatin (IC50=29.8µM). Furthermore, compounds 10b, 10c, 11a, 11c, and 11d demonstrated a significant cytotoxic potential against the MCF-7 cancer cell line with a single-digit micromolar IC50 potency (4.8-8.5µM) compared to the standard drug paclitaxel (IC50=19.7µM). The cytotoxic studies of above selected potent active hybrid compounds, against HEK-293, normal cell line, further highlight the potential use of 10c molecule (IC50=4.8µM against MCF-7 cells) as an anticancer agent for breast cancer with a selectivity index of 2.597. The cytotoxic results were further supported by the molecular docking studies.

Novel Pyrimidine Tethered Benzamide Derivatives as Potential Anticancer Agents: Synthesis, Characterization, Molecular Docking and In vitro Cytotoxicity Evaluation

Current research work presents the synthesis, characterization, molecular docking study and in vitro cytotoxicity evaluation of novel pyrimidine-tethered benzamide derivatives as potential anticancer agents. The synthesis involved multi-steps procedure, including the synthesis of various chalcones (3a-t) with corresponding ketones (1 and 2) and substituted aldehydes (a-j), followed by pyrimidine amines (5a-t) with condensation of chalcones and guanidine and finally pyrimidinyl benzamide derivatives (8a-t) from compounds 5a-t coupling with acid chloride (7) using DIBAL-H. Synthesized compounds characterized by NMR spectroscopy and HRMS. The molecular docking studies were conducted against EGFR (6LUD) and CDK-4 (7SJ3) receptors, revealing distinct binding affinities influenced by substituent groups. Additionally, the cytotoxicity of the synthesized compounds was evaluated using the MTT assay against various cancer cell lines, including A-549 (non-small cell lung cancer), HCT-116 (colorectal cancer), PANC-1 (pancreatic cancer) and HaLa (cervical cancer), along with one normal human embryonic kidney cell line (HEK-293). Among the synthesized compounds, derivatives 8f and 8j exhibited the best anticancer activity against A-549 cells, compounds 8j and 8e showed exceptional potency against HCT-116 cells, compounds 8f and 8j demonstrated high efficacy against PANC-1 cells and compound 8h displayed remarkable potency against HaLa cells. The findings highlight the potential of these pyrimidinyl benzamide derivatives as targeted anticancer agents.

Synthesis and Evaluation of 3,5-Disubstituted-1,2,4-Oxadiazolyl Benzamides as Potential Anti-Breast Cancer Agents: In Vitro and In Silico Studies.

Herein, the synthesis, anti-cancer evaluation, and in silico studies of a series of 1,2,4-oxadiazole compounds (8-15) are disclosed. The synthesized molecules were tested in vitro for anti-cancer activity against MCF-7, MDA-MB-231, HeLa, Ishikawa cell lines and human embryonic kidney (HEK-293) cell lines. Among the synthesized compounds, 9 and 15 exhibited significant cytotoxicity, with IC50 values of 7.82 μM and 6.02 μM, respectively, against MCF-7 cell line, better than that of anti-breast cancer drug, tamoxifen (IC50=11.92 μM), used as control. Significantly, both 9 and 15 exhibited very low toxicity (IC50>20 μM) against normal HEK-293 cells. This suggests them as potentially effective anti-cancer lead molecules. The in vitro anti-cancer data was supported by in silico studies which also identified compounds 9 and 15 as potent inhibitors of the 17β-hydroxysteroid dehydrogenase1 (17β-HSD1) proteins, demonstrating strong interactions and stability The atom-based QSAR model exhibited high accuracy, significant regression, and predictive reliability, aiding in understanding and optimizing biological activity. The drug-likeness study of compounds 9 and 15 indicated favorable pharmacokinetics, with in silico toxicity predictions showing compound 15 to be non-toxic. These findings suggest compounds 9 and 15 as potential lead molecules against breast cancer.

Investigation of the cytotoxic activity, DFT calculation, and docking studies newly synthesized 1,3-disubstituted benzimidazolium chlorides on human liver cancer, lung cancer, and normal embryonic kidney cell lines

Three 1,3-disubstituted benzimidazolium salts (3a-c) were efficiently synthesized in moderate to high yields (52–83 %) and analyzed through NMR spectra. The anti-cancer efficiency of these compounds was tested on human liver cancer (HepG2), lung cancer (A549), and normal embryonic kidney (HEK-293T) cell lines. The results demonstrated that compound 3b emerges as a promising candidate for further investigation due to its high cytotoxicity, comparable to cisplatin. The optimized geometry, electronic properties, chemical parameters and frontier molecular orbitals of 3a-c were determined by DFT calculation using density functional theory (DFT) with the B3LYP/6–31++G(d,p) level in the ground state. In addition to the experimental studies, compounds 3a-c were docked against target protein PDB ID: 6V9C representing the HepG2 cell line.

Polydopamine-Mediated Antimicrobial Lipopeptide Surface Coating for Medical Devices.

Biofilm formation on medical implants such as catheters is a major issue which needs to be addressed as it leads to severe health care associated infections. This study explored the design and synthesis of a polydopamine-lipopeptide based antimicrobial coating. The coating was used to modify the surface of Ultrathane Catheters. The lipopeptide SL1.15 with an N-terminal cysteine was covalently conjugated to the polydopamine modified catheters via a Michael addition reaction between the thiol moiety in the peptide and the aromatic ring in the polydopamine layer. The immobilization of the peptide on the polydopamine coated catheters was confirmed using water contact angle, X-ray photoelectron spectroscopy, atomic force microscopy, and scanning electron microscopy (SEM). The antimicrobial activity of the coated catheters investigated using drug resistant and clinical strains of Gram-positive (MRSA and S. aureus) and Gram-negative (E. coli, A. baumannii, and P. aeruginosa) bacteria revealed that lipopeptide immobilization inhibited >90% bacterial adhesion to the catheter surface. Additionally, biofilm assays against MRSA and E. coli revealed that the lipopeptide immobilized catheters inhibited >85% bacterial growth after 1 week incubation. Finally, the cytotoxicity profile of the catheters using the human dermal fibroblast, and the human embryonic kidney cell lines demonstrated that the polydopamine-lipopeptide coating was not toxic after 72 h incubation.

Novel bioassays based on 3D-printed device for sensing of hypoxia and p53 pathway in 3D cell models.

Cell-based assays are widely exploited for drug screening and biosensing, providing useful information about bioactivity of target analytes and complex biological samples. It is well recognized that 3D cell models are required to achieve highly valuable information, also from the perspective of replacing animal models. However, bioassays relying on 3D cell models are generally highly demanding in terms of facilities, equipment, and skilled personnel requirements. To reduce cost, increase sustainability, and provide a flexible 3D cell-based platform for bioassays, we here report a novel approach based on a 3D-printed microtissue device. To assess the suitability of this strategy for reporter gene technology, we selected to monitor two molecular pathways which were of interest in several applications, hypoxia signaling and the p53 pathway. The investigation of such pathways is highly relevant in fields spanning from drug screening to bioactivity monitoring for industrial by-product valorization. Microtissues of human hepatocarcinoma (HepG2) and human embryonic kidney (Hek293T) cell lines were obtained with a low-cost and sustainable chip platform and bioassays were developed to monitor the hypoxia-inducible factors (HIFs) and the p53 tumor suppressor pathway. HepG2 and Hek293T 3D cell models were genetically engineered to express the Luc2P from Photinus pyralis firefly either under the regulation of p53 or HIF response elements. The bioassays allowed quantitative assessment of hypoxia and tumoral activity with 1,10-phenanthroline for HIF and with doxorubicin for p53 pathway activation, respectively, showing good potential for applications of this sustainable and low-cost 3D-printed microfluidic platform for bioactivity analyses, drug screening, and precision medicine.

What is the real value of omics data? Enhancing research outcomes and securing long-term data excellence.

A wealth of high-throughput biological data, of which omics constitute a significant fraction, has been made publicly available in repositories over the past decades. These data come in various formats and cover a range of species and research areas providing insights into the complexities of biological systems; the public repositories hosting these data serve as multifaceted resources. The potentially greater value of these data lies in their secondary utilization as the deployment of data science and artificial intelligence in biology advances. Here, we critically evaluate challenges in secondary data use, focusing on omics data of human embryonic kidney cell lines available in public repositories. The emerging issues are obstacles faced by secondary data users across diverse domains as they concern platforms and repositories, which accept deposition of data irrespective of their species type. The evolving landscape of data-driven research in biology prompts re-evaluation of open access data curation and submission procedures to ensure that these challenges do not impede novel research opportunities through data exploitation. This paper aims to draw attention to widespread issues with data reporting and encourages data owners to meticulously curate submissions to maximize not only their immediate research impact but also the long-term legacy of datasets.

7-(Substituted amino)-5-methylthioazolo[1,5-a]pyrimidines: Synthesis, cytotoxic properties in vitro and molecular docking

A method for the synthesis of new 7-(substituted amino)-5-methylthioazolo[1,5-a]pyrimidines has been developed. Based on the MTT test, IC50 values were calculated for the obtained compounds against lung carcinoma (A549), liver carcinoma (HepG2), embryonal rhabdomyosarcoma (Rd) and human embryonic kidney (HEK 293) cell lines. Some compounds from the series demonstrated activity close to the reference drug, but with a certain selectivity. Based on the results of MTT assay and molecular docking studies for the catalytic subunits of PI3K, two isoforms (PI3Kβ and PI3Kδ) were assumed as the targets for the new series of azolo[1,5-a]pyrimidines with cytotoxic effect on the rhabdomyosarcoma cell line.

Optimization of Exon-Skipping Riboswitches and Their Applications to Control Mammalian Cell Fate.

Mammalian riboswitches that can regulate transgene expression via RNA-small molecule interaction have promising applications in medicine and biotechnology, as they involve no protein factors that can induce immunogenic reactions and are not dependent on specially engineered promoters. However, the lack of cell-permeable and low-toxicity small molecules and cognate aptamers that can be exploited as riboswitches and the modest switching performance of mammalian riboswitches have limited their applications. In this study, we systematically optimized the design of a riboswitch that regulates exon skipping via an RNA aptamer that binds ASP2905. We examined two design strategies to modulate the stability of the aptamer base stem that blocks the 5' splice site to fine-tune the riboswitch characteristics. Furthermore, an optimized riboswitch was used to generate a mouse embryonic stem cell line that can be chemically induced to differentiate into myogenic cells by activating Myod1 expression and a human embryonic kidney cell line that can be induced to trigger apoptosis by activating BAX expression. The results demonstrate the tight chemical regulation of transgenes in mammalian cells to control their phenotype without exogenous protein factors.

Salmonella Detection in Food Using a HEK-hTLR5 Reporter Cell-Based Sensor.

The development of a rapid, sensitive, specific method for detecting foodborne pathogens is paramount for supplying safe food to enhance public health safety. Despite the significant improvement in pathogen detection methods, key issues are still associated with rapid methods, such as distinguishing living cells from dead, the pathogenic potential or health risk of the analyte at the time of consumption, the detection limit, and the sample-to-result. Mammalian cell-based assays analyze pathogens' interaction with host cells and are responsive only to live pathogens or active toxins. In this study, a human embryonic kidney (HEK293) cell line expressing Toll-Like Receptor 5 (TLR-5) and chromogenic reporter system (HEK dual hTLR5) was used for the detection of viable Salmonella in a 96-well tissue culture plate. This cell line responds to low concentrations of TLR5 agonist flagellin. Stimulation of TLR5 ligand activates nuclear factor-kB (NF-κB)-linked alkaline phosphatase (AP-1) signaling cascade inducing the production of secreted embryonic alkaline phosphatase (SEAP). With the addition of a ρ-nitrophenyl phosphate as a substrate, a colored end product representing a positive signal is quantified. The assay's specificity was validated with the top 20 Salmonella enterica serovars and 19 non-Salmonella spp. The performance of the assay was also validated with spiked food samples. The total detection time (sample-to-result), including shortened pre-enrichment (4 h) and selective enrichment (4 h) steps with artificially inoculated outbreak-implicated food samples (chicken, peanut kernel, peanut butter, black pepper, mayonnaise, and peach), was 15 h when inoculated at 1-100 CFU/25 g sample. These results show the potential of HEK-DualTM hTLR5 cell-based functional biosensors for the rapid screening of Salmonella.

Production of a magnetic nanocomposite for biological and hyperthermia applications based on chitosan-silk fibroin hydrogel incorporated with carbon nitride

Hydrogels based on natural polymers have lightened the path of novel drug delivery systems, wound healing, and tissue engineering fields because they are renewable, non-toxic, biocompatible, and biodegradable. Furthermore, applying modified hydrogels can upgrade their biological activity. Herein, Chitosan (CS) was used to create a hydrogel using terephthaloyl thiourea as a cross-linker. Silk fibroin (SF) and carbon nitride (CN) were added to the hydrogel to enhance its strength and biocompatibility. Finally, CS hydrogel/SF/CN was in situ magnetized using Fe3O4 magnetic nanoparticles (MNPs) and manufactured as a nanobiocomposite for improved hyperthermia. The structural properties of the nanobiocomposite were assessed using several analytical techniques, including VSM, FTIR, TGA, EDX, XRD, and FESEM. The saturation magnetization of this magnetic nanocomposite was 23.94 emu/g. The hemolytic experiment on the nanobiocomposite resulted in ca. 98 % cell survival, with a hemolysis rate of 1.69 %. Anticancer property is confirmed by a 20.0 % reduction in cell viability of BT549 cells at 1.75 mg/mL concentration compared to 0.015 mg/mL. The nanocomposite is non-toxic to the human embryonic kidney cell line (HEK293T), indicating its potential for biomedical applications. Finally, the magnetic nanocomposite's hyperthermia behavior was examined using a specific absorption rate (SAR), achieving the highest value of 47.44 W/g at 200.0 kHz. When subjected to an alternating magnetic field, the nanobiocomposite may perform well in hyperthermia therapy. These results indicate that the magnetic nanobiocomposite has the potential to perform well in hyperthermia therapy when subjected to an alternating magnetic field.

Effect of silver nanochitosan on control of seed-borne pathogens and maintaining seed quality of wheat

Seeds, considered as the foundation of agriculture, are invaded by a broad spectrum of seed-borne pathogens. The current study aimed to control seed-borne fungal pathogens of wheat, Aspergillus flavus and A. niger, by using Ag+ nanochitosan (Ag-NC) for nano-priming of seeds and enhancing seed quality. Nanochitosan (NC) and Ag-NC were synthesized using the gelation method and characterized by UV–vis spectrophotometry, FESEM, EDXS, and HRTEM. NC and Ag-NC showed irregular surface topography with an average particle size of 275 and 325 nm, respectively. Antifungal activity of both the nanoparticles at 0.1, 0.2, 0.3, 0.4, and 0.5 mg/mL revealed that Ag-NC at 0.5 mg/mL has completely terminated the mycelial growth of both pathogens. Malonaldehyde content increased to 77.77% in A. flavus and 82.66% in A. niger when exposed to 0.5 mg/mL Ag-NC. High-intensity fluorescence due to oxidative stress was observed in Ag-NC-treated pathogens. Ultra-structural changes in Ag-NC treated pathogenic spores under SEM displayed pronounced membrane damages. Wheat seeds were nano-primed with NC and Ag-NC at 0.5 mg/mL, and fungal load was examined to evaluate the mitigation of pathogenic stress and its effect on seedling growth promotion activity. Ag-NC priming reduced the fungal load and allowed successful seed germination. Ag-NC priming increased the albumin, gliadin, gluten, and glutenin content along with total phenol, reducing sugar and starch levels. Ag-NC priming increased the overall protein levels traced through SDS-PAGE. Seed priming with Ag-NC promotes seed germination, mean germination time, stress tolerance index, vigour, etc. NC and Ag-NC at 0.5 mg/mL showed no cytotoxic effect on the Human Embryonic Kidney (HEK293) cell line that ensures the nanoparticles are non-toxic. Thus, the synthesized nanoparticles exhibit a dual role in antifungal activity and plant growth promotion.

Recombinant human epidermal growth factor-loaded liposomes and transferosomes for dermal delivery: Development, characterization, and cytotoxicity evaluation.

Recombinant human epidermal growth factor (rhEGF) is widely utilized as an antiaging compound in wound-healing therapies and cosmetic purposes. However, topical administration of rhEGF has limited treatment outcomes because of its poor percutaneous penetration and rapid proteinase degradation. To overcome these obstacles, this study aims to develop and characterize rhEGF-containing conventional liposomes (rhEGF-CLs) and transferosomes (rhEGF-TFs) as efficient dermal carriers. Physicochemical characterization such as particle size, zeta potential (ZP), morphology, encapsulation efficiency (EE%), and release properties of nanocarriers as well as in vitro cytotoxicity in human dermal fibroblast (HDF) and human embryonic kidney (HEK293) cell lines were investigated. rhEGF-TFs at the rhEGF concentration ranging from 0.05 to 1.0 μg/mL were chosen as the optimum formulation due to the desired release profile, acceptable EE%, optimal cell proliferation, and minimal cytotoxicity compared to the control and free rhEGF. However, higher concentrations caused a decrease in cell viability. The ratio 20:80 of Tween 80 to lipid was optimal for rhEGF-TFs-2, which had an average diameter of 233.23 ± 2.64 nm, polydispersity index of 0.33 ± 0.05, ZP of -15.46 ± 0.29 mV, and EE% of 60.50 ± 1.91. The formulations remained stable at 5°C for at least 1 month. TEM and SEM microscopy revealed that rhEGF-TFs-2 had a regular shape and unilamellar structure. In vitro drug release studies confirmed the superiority of rhEGF-TFs-2 in terms of optimal cumulative release of rhEGF approximately 82% within 24 h. Franz diffusion cell study showed higher rhEGF-TFs-2 skin permeation compared to free rhEGF solution. Taken together, we concluded that rhEGF-TFs can be used as a promising formulation for wound healing and skin regeneration products.

DETERMINATION OF THE CIRCADIAN OSCILLATION PATTERN OF UNFOLDED PROTEIN RESPONSE SIGNALING COMPONENTS IN HUMAN EMBRYONIC KIDNEY HEK293 CELLS

Objective: The circadian rhythm is one of the primary regulatory systems with near 24-hour oscillations. It has a crucial role in regulating physiological conditions in the human body, including body temperature and the secretion of hormones. Numerous disorders, such as cancer and diabetes, have been linked to disruptions of the cellular circadian rhythm. Herein, we aimed to investigate the relationship between the circadian rhythm and unfolded protein response (UPR) signaling, which is one of the important physiological mechanisms in mammalian cells and has recently been associated with drug resistance, invasion and metastasis in cancer. Material and Method: Human embryonic kidney cell line HEK293 was provided from the American Type Culture Collection and propagated in DMEM containing 10% FBS and growth ingredients. For in vitro circadian synchronization, cells were exposed to 50% and then the oscillation pattern of gene and protein expression of UPR-related target genes was analyzed by agarose gel electrophoresis and immunoblotting, respectively. The oscillation pattern was commented on through curve-fitting analysis. Result and Discussion: Our findings demonstrated that UPR components, including IRE1α, XBP-1s, eIF2α, phospho(Ser51)-eIF2α, PERK, ATF4, GADD34 and ATF6, tightly exhibit oscillation patterns under a circadian rhythm on a 48-hour time scale like the PER1 gene that is a core component of the circadian rhythm. Moreover, endoplasmic reticulum (ER) stress genes, BiP/GRP78 and CHOP, were similar to UPR components under the circadian rhythm. Additionally, we found the activation of UPR signaling harmoniously modulated with the circadian rhythm. Present data indicated that the expression level of UPR components exhibited strict oscillation under the circadian rhythm. Our findings may guide experimental studies of new-generation UPR-targeted drugs to be developed to treat various pathologies in accordance with the circadian rhythm.

Synthesis, spectral, in silico/molecular docking and pharmacological studies of biologically potent triorganotin(IV) carboxylates

A series of compounds containing a glutaric anhydride-based carboxylate ligand (LH) and three triorganotin derivatives R3SnL for R = Me (1), nBu (2), and pH (3) are described in this article. The synthesis of the compounds was confirmed by microelemental CHN analysis, FT-IR, NMR and in the case of LH, 1 and 2 by single crystal X-ray crystallography, revealing the formation of one-dimensional coordination polymers for 1 and 2 with trans-C3O2 trigonal-bipyramidal geometries for the tin(IV) atoms. The anti-microbial activity was explored with the MIC values of 2 (≤ 0.25 µg/mL against the bacterium S. aureus and fungal strains C. albicans and C. neoformans) being less than observed for the anti-bacterial drug vancomycin-HCl (1 μg/mL) and anti-fungal drug fluconazole (≤ 0.25 and 8 μg/mL, respectively). The maximum cytotoxicity was observed for 2, against the human embryonic kidney (HEC239) cell line, with CC50 ≤ 25 μg/mL. An evaluation of the compound-DNA interactions was performed through UV–Visible spectroscopic and viscometric experiments, with the observation of hypochromic along with slight bathochromic shifts and an increase in viscosity, respectively, revealing an intercalative mode of interaction. In comparison with the standard anti-oxidant ascorbic acid, the greatest scavenging of DPPH was recorded for 3 followed by 2, 1 and LH. The human malignant glioma cell line (MG-U87) was used to explore the anti-cancer potential of the synthesised compounds. Compounds 3 and 2 (IC50 = 5 and 21 µM, respectively) showed encouraging results when compared to cisplatin (IC50 = 133 µM). The anti-leishmanial data shows that at a 1000 µg/mL dose, the greatest percentage inhibition was observed for 2 (87.15 ± 0.01 %) which is almost equivalent to that for amphotericin B (90.67 ± 0.15 %). The docking results with SS-DNA revealed the compounds undergo an intercalative mode of interaction with the nitrogenous bases adenine, guanine and cytosine, with docking score order: 3 (-8.11) > 2 (-8.02) > 1 (-6.99) > LH (-6.38). Similarly, the docking results against the Brest cancer receptor BRCA2, indicate the highest binding score was observed for 3. The anti-leishmanial docking data indicates 2 as having maximum docking score (-7.79). The in silico study, performed to reveal the drug-likeness behaviour of the news compounds, suggest that they possess the properties of orally active drugs.

NanoTrans: an integrated computational framework for comprehensive transcriptome analysis with nanopore direct RNA sequencing

Nanopore direct RNA sequencing (DRS) provides the direct access to native RNA strands with full-length information, shedding light on rich qualitative and quantitative properties of gene expression profiles. Here with NanoTrans, we present an integrated computational framework that comprehensively covers all major DRS-based application scopes, including isoform clustering and quantification, poly(A) tail length estimation, RNA modification profiling, and fusion gene detection. In addition to its merit in providing such a streamlined one-stop solution, NanoTrans also shines in its workflow-orientated modular design, batch processing capability, all-in-one tabular and graphic report output, as well as automatic installation and configuration supports. Finally, by applying NanoTrans to real DRS datasets of yeast, Arabidopsis, as well as human embryonic kidney and cancer cell lines, we further demonstrate its utility, effectiveness, and efficacy across a wide range of DRS-based application settings.