Human Embryonic Kidney Cell Line Research Articles

Oncogenic Cells of Renal Embryonic Lineage Sensitive to the Small-Molecule Inhibitor QC6352 Display Depletion of KDM4 Levels and Disruption of Ribosome Biogenesis.

The histone lysine demethylases KDM4A-C are involved in physiologic processes including stem cell identity and self-renewal during development, DNA damage repair, and cell-cycle progression. KDM4A-C are overexpressed and associated with malignant cell behavior in multiple human cancers and are therefore potential therapeutic targets. Given the role of KDM4A-C in development and cancer, we aimed to test the potent, selective KDM4A-C inhibitor QC6352 on oncogenic cells of renal embryonic lineage. The anaplastic Wilms tumor cell line WiT49 and the tumor-forming human embryonic kidney cell line HEK293 demonstrated low nanomolar QC6352 sensitivity. The cytostatic response to QC6352 in WiT49 and HEK293 cells was marked by induction of DNA damage, a DNA repair-associated protein checkpoint response, S-phase cell-cycle arrest, profound reduction of ribosomal protein gene and rRNA transcription, and blockade of newly synthesized proteins. QC6352 caused reduction of KDM4A-C levels by a proteasome-associated mechanism. The cellular phenotype caused by QC6352 treatment of reduced migration, proliferation, tumor spheroid growth, DNA damage, and S-phase cell-cycle arrest was most closely mirrored by knockdown of KDM4A as determined by siRNA knockdown of KDM4A-C. QC6352 sensitivity correlated with high basal levels of ribosomal gene transcription in more than 900 human cancer cell lines. Targeting KDM4A may be of future therapeutic interest in oncogenic cells of embryonic renal lineage or cells with high basal expression of ribosomal protein genes.

Antioxidant and Cytotoxic Activities of Leaf and Stem Extracts of Barleria albostellata C.B. Clarke

Barleria albostellata C.B. Clarke (Acanthaceae) is a plant native to South Africa and relatively few studies have been performed on it. Species in this genus are known for their ethnopharmacological and phyto-medicinal values. In this study, the total flavonoid and phenolic contents and the antioxidant and cytotoxic activities of hexane, chloroform, and methanol extracts were evaluated at five different concentrations (15, 30, 60, 120, and 240 μg/mL). The antioxidant activity of the extracts of B. albostellata was assessed in vitro using the 2,2′-diphenyl-1-picrylhydrazyl (DPPH) scavenging and ferric reducing antioxidant power (FRAP) assays, while the phenolic content was determined using a Folin–Ciocalteu assay. The extracts’ cytotoxicity was established using a 3-[(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] (MTT) assay in human embryonic kidney (HEK293), cervical cancer (HeLa), and breast adenocarcinoma (MCF-7) cell lines. Methanolic leaf extracts had the highest total flavonoid content (42.39 ± 1.14 mg GAE/g DW) compared to other solvents. Additionally, the total phenolic content was the greatest in the methanol leaf extract (6.05 ± 0.09 GAE/g DW), followed by the methanol stem extracts (2.93 ± 0.73 GAE/g DW). The methanolic leaf and stem extract concentrations needed for 50% inhibition (DPPH) were 16.95 µg/mL and 14.27 µg/mL, respectively, whereas for FRAP, the reducing powers of all extracts were considerably lower than the ascorbic acid standard. The IC50 values of extracts tested in the three cell lines were >63 µg/mL. According to the findings of our study, the leaves and stems of B. albostellata are rich in several bioactive compounds that may be a possible source of natural antioxidants and may have the potential to treat certain diseases. The extraction of the bioactive compounds from the leaves and stems of B. albostellata using bioassay-guided fractionation and the assessment of their safety will be essential for further investigations into this species in the search of potential novel therapeutic drug leads. To the best of our knowledge, this is the first report of the cytotoxic activities of leaf and stem extracts of Barleria albostellata.

In vitro and in silico guided identification of antimalarial phytoconstituent(s) in the root of Citrus maxima (Burm.) Merr

As a part of our continuous effort to find new therapeutic agents from natural sources, the hydroalcoholic (1:1) extract of Citrus maxima (Burm.) Merr. root was selected for the identification of possible antimalarial phytoconstituents. From the extract, three flavonoids including luteolin were isolated and evaluated for in vitro antimalarial activity against the chloroquine-sensitive (Pf3D7) and resistant (PfRKL-9) strains of Plasmodium falciparum. Among these, luteolin (CM3) showed the highest antimalarial activity with IC50 values of 2.315 ± 0.489 and 2.691 ± 0.454 µg/ml against the Pf3D7 and PfRKL-9 strains respectively. To assess the safety of luteolin (CM3), a cytotoxicity study against a normal human embryonic kidney cell line (HEK-293) was performed and the compound was found to be safe with a CC50 value of 222.3 ± 1.443 µg/ml. The docking study against 26 target proteins of P. falciparum revealed that luteolin (CM3) has a better binding affinity with two proteins, viz. P. falciparum lactate dehydrogenase (PfLDG) and P. falciparum enoyl-ACP reductase (PfEAR) in comparison to the co-crystallized ligands. Furthermore, the molecular dynamics simulation study of the protein-ligand complexes also supported the binding affinity and interactions of luteolin (CM3) at the active sites. Finally, the binding free energy calculation revealed that the luteolin formed a thermodynamically more stable complex with PfLDG (-50.955 ± 17.184 kJ/mol) than PfEAR (-24.856 ± 13.739 kJ/mol). Overall, in this study, we identified an antimalarial marker in the hydroalcoholic extract of C. maxima root which may act by inhibiting PfLDG. Communicated by Ramaswamy H. Sarma

Identification of m6A-associated genes as prognostic and immune-associated biomarkers in Wilms tumor

ObjectivesWilms tumor (WT) is a common renal malignant tumor in children. We aimed to investigate the potential prognostic value of m6A-related genes and their relationship to the immune microenvironment in WT.MethodsRNA-seq data and clinical information from 121 WT and 6 normal samples were obtained from the University of California Santa Cruz Xena database. We used various bioinformatics analysis tools to analyze these data and verify the expression level of m6A-related genes by experiments.ResultsFour m6A-related genes were successfully screened, including ADGRG2, CPD, CTHRC1, and LRTM2. Kaplan–Meier survival curves showed that the four genes were closely related to the prognosis of WT, which was also confirmed by receiver operator characteristic curves. Subsequently, in the immune microenvironment of WT, we discovered that Th1_cells were positively correlated with ADGRG2, CCR was negatively correlated with CPD, CCR was positively correlated with CTHRC1, APC_co_stimulation, CCR, Macrophages, inflammation-promoting cells, Treg, and Type_II_IFN_Reponse were negatively correlated with LRTM2. Finally, qRT-PCR showed that expression levels of the four genes were upregulated in the nephroblastoma cell lines (G-401, SK-NEP-1, and WT-CLS1) compared with the human embryonic kidney cell lines (293T).ConclusionsTaken together, our study first time screened the m6A-related genes and revealed that ADGRG2, CPD, CTHRC1, and LRTM2 are the prognostic and immune-associated biomarkers in WT.

Experimental and theoretical confirmation of CeFeCu trimetal oxide/Bi2O3 S-scheme heterojunction for boosted photocatalytic degradation of sulfamethoxazole and toxicity evaluation

Photocatalytic degradation of pharmaceutical pollutants in water or wastewater is becoming a hot topic for environmental clean-up. Herein, a novel CeFeCu trimetal oxide (CFC TMO)/Bi2O3 (CFCB) photocatalysts were prepared through precursor mixing, ultrasonication, and calcination methods and used for photocatalytic degradation of sulfamethoxazole (SMZ). The pseudo-first order kinetic rate of CFCB-2 catalyst on degradation of SMZ reached 0.0577 min−1 which is 4.93 and 10.02 times higher than of α-Bi2O3 and CFC TMO under visible light illumination. This was proved by the strong interaction between the pristine materials as confirmed by XPS and density functional theory (DFT) calculations. DFT calculations indicated that having more orbital distribution could make it easier to capture more electrons and produce more radicals through electron transfer. Meanwhile SMZ by-products showed no sign of cytotoxicity towards human embryonic kidney cell line (HEK 293) and human intestinal epithelial cell line (CACO-2). The findings of the scavenger experiments and ESR measurements proved that •OH and •O2− radicals played superior role in decomposition of SMZ. Furthermore, the S-scheme charge carrier transfer and degradation pathway of SMZ are proposed and analysed. This study paves ways into promising strategy for the construction of α-Bi2O3 based novel S-scheme heterojunction for environmental decontamination and can be extended to other applications such as Fenton-like reactions and persulfate activation.

Green extraction of berberine from Coscinium fenestratum (Gaertn.) Colebr. using ultrasound-assisted aqueous solutions of organic acids, polyalcohols, and deep eutectic solvents

Berberine displays many potential bioactivities and is found in several medicinal herbs, such as Coscinium fenestratum. The extraction of berberine from biomass was carried out using volatile organic solvents, inorganic acid solutions, and more recently, ionic liquids and deep eutectic solvents. In order to develop more efficient and sustainable extraction processes, this study used inexpensive green solvents, such as carboxylic acid and polyalcohol aqueous solutions, to extract berberine from Coscinium fenestratum. Initial screening was performed to compare the berberine extraction performance of various solvents, including aqueous solutions of carboxylic acids, polyalcohols, and their deep eutectic solvents. The obtained results showed that lactic acid was a potential solvent for extracting berberine. After optimizing the process using response surface methodology (RSM), the optimal ultrasound-assisted extraction conditions were determined to be a lactic acid concentration of 60 % (w/w), a liquid-to-solid ratio of 17.25 mL/g, an extraction time of 20 min, and an extraction temperature of 66 °C. This resulted in an extraction efficiency of 38.23 ± 0.69 mg/g, corresponding to a berberine concentration of 2.22 ± 0.04 mg/mL. Additionally, the use of lactic acid as the extraction solvent yielded superior extraction efficiency and berberine purity compared to conventional volatile organic solvents, and inorganic acids, bases. The recovery of berberine was achieved using macroporous resins, allowing for the recovery of 99.52 ± 0.67 % of berberine from aqueous lactic acid solutions. The lactic acid solvent and the AB-8 resin were successfully recovered and reused for a minimum of three consecutive extraction cycles, maintaining their full effectiveness throughout the entire process. The recovered berberine from the lactic acid extract exhibited stronger antibacterial activity than the extracts obtained using ethanol, methanol, and acetone, as well as berberine recovered from the limewater and sulfuric acid extracts. Furthermore, the lactic acid solvent and its extracts demonstrated no cytotoxicity toward the human embryonic kidney cell line. The findings of this investigation establish a foundation for future studies utilizing eco-friendly solvents for alkaloid extraction from medicinal plants. This lays the groundwork for implementing a large-scale extraction process of natural active compounds in a safe and environmentally conscious manner.

Anisomeles indica Extracts and Their Constituents Suppress the Protein Expression of ACE2 and TMPRSS2 In Vivo and In Vitro.

Coronavirus disease 2019 (COVID-19), stemming from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has had a profound global impact. This highly contagious pneumonia remains a significant ongoing threat. Uncertainties persist about the virus's effects on human health, underscoring the need for treatments and prevention. Current research highlights angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2) as key targets against SARS-CoV-2. The virus relies on ACE2 to enter cells and TMPRSS2 to activate its spike protein. Inhibiting ACE2 and TMPRSS2 expression can help prevent and treat SARS-CoV-2 infections. Anisomeles indica (L.) Kuntze, a medicinal plant in traditional Chinese medicine, shows various promising pharmacological properties. In this study, ethanolic extracts of A. indica were examined both in vivo (250 and 500 μM) and in vitro (500 μM). Through Western blotting analysis, a significant reduction in the expression levels of ACE2 and TMPRSS2 proteins was observed in HepG2 (human hepatocellular carcinoma) cells and HEK 293T (human embryonic kidney) cell lines without inducing cellular damage. The principal constituents of A. indica, namely, ovatodiolide (5 and 10 μM), anisomlic acid (5 and 10 μM), and apigenin (12.5 and 25 μM), were also found to produce the same effect. Furthermore, immunohistochemical analysis of mouse liver, kidney, and lung tissues demonstrated a decrease in ACE2 and TMPRSS2 protein expression levels. Consequently, this article suggests that A. indica and its constituents have the potential to reduce ACE2 and TMPRSS2 protein expression levels, thus aiding in the prevention of SARS-CoV-2 infections.

Biomedical potential of hydrothermally synthesized zinc oxide nanoparticles for antifungal evaluation and cytotoxicity analysis

As nanotechnology gains popularity in medicine, there is a growing concern that small nanoparticles (NPs) could accumulate and penetrate deep into the human body, posing potential risks to various organs. To ensure safe usage and assess potential hazards, it is crucial to conduct a toxicological evaluation of ZnO NPs on human organs such as the kidney. The present study involved the synthesis of ZnO NPs by using the hydrothermal method. Pure and crystalline phase was confirmed by X‐ray diffraction (XRD). The SEM images displayed less agglomerated surface morphology, implying that the NPs were distributed relatively evenly and exhibited a plate‐like shape. Two pathogenic fungi Fusarium incarnatum and Aspergillus niger were isolated from the capsicum and tomato, respectively, and identified through morphological and molecular analysis. The antimycotic property of synthesized ZnO NPs were examined against Fusariumincarnatum (MT682502) and Aspergillus niger (MT675916) and showed decreased microbial growth after 9 days of incubation. The toxicological evaluation of ZnO NPs on a human embryonic kidney (HEK293) cell line showed 88% cell viability with 10 μg/ml, leading to the safe use of ZnO NPs.

Green synthesis of selenium nanoparticles using Artemisia chamaemelifolia: toxicity effects through regulation of gene expression for cancer cells and bacteria

An effective and economical green procedure is reported for the synthesis of selenium nanoparticles (SeNPs) using the extract of Artemisia chamaemelifolia aerial. The as-synthesized SeNPs were characterized using different spectroscopy and microscopy techniques, confirming the formation of about 110 nm SeNPs capped with phytochemicals present in the Artemisia chamaemelifolia aerial extract. The cytotoxicity of the as-synthesized SeNPs was evaluated through their effects on different bacteria and cell lines. The cytotoxicity effect was tested against the tumorigenic human colorectal adenocarcinoma (HT-29) cell line as well as the low-tumorigenic human embryonic kidney (HEK293) cell line. The results suggested that there is less cytotoxicity against HEK293 cells as compared to HT-29 cells. The apoptosis mechanism was further evaluated by investigation of different anti-apoptotic or pro-apoptotic genes expressed in these two cell lines. For the bacterial toxicity evaluation, S. aureus and K. pneumoniae were used. The results suggested considerable toxicity effect against both studied strains. Similarly, the expression levels of hlaA and vgrG-1 genes were evaluated in pathogenic bacteria after treatment with SeNPs. It was found that SeNPs could down-regulate the expression of pathogenic genes (hlaA and vgrG-1 genes in S. aureus and K. pneumoniae strains, respectively). Based on the findings of this study, we believe SeNPs synthesized by plant extract have great potential to be used for biomedical applications and such a non-metal nanoparticle will open new avenues to future medical applications as an anticancer and antibacterial agent.

Application of Iron Nanoparticles Synthesized from a Bioflocculant Produced by Yeast Strain Pichia kudriavzevii Obtained from Kombucha Tea SCOBY in the Treatment of Wastewater.

Studying the production of Iron (Fe) nanoparticles using natural substances is an intriguing area of research in nanotechnology, as these nanoparticles possess biocompatibility and natural stability, which make them useful for a variety of industrial applications. The study utilized Fe nanoparticles that were synthesized using a bioflocculant and applied to eliminate different kinds of pollutants and dyes found in wastewater and solutions. The study involved the generation of Fe nanoparticles through a bioflocculant obtained from Pichia kudriavzevii, which were evaluated for their flocculation and antimicrobial capabilities. The impact of the Fe nanoparticles on human embryonic kidney (HEK 293) cell lines was studied to assess their potential cytotoxicity effects. An array of spectroscopic and microscopic methods was employed to characterize the biosynthesized Fe nanoparticles, including SEM-EDX, FT-IR, TEM, XRD, UV-vis, and TGA. A highly efficient flocculating activity of 85% was achieved with 0.6 mg/mL dosage of Fe nanoparticles. The biosynthesized Fe nanoparticles demonstrated a noteworthy concentration-dependent cytotoxicity effect on HEK 293 cell lines with the highest concentration used resulting in 34% cell survival. The Fe nanoparticles exhibited strong antimicrobial properties against a variety of evaluated Gram-positive and Gram-negative microorganisms. The efficiency of removing dyes by the nanoparticles was found to be higher than 65% for the tested dyes, with the highest being 93% for safranine. The Fe nanoparticles demonstrated remarkable efficiency in removing various pollutants from wastewater. In comparison to traditional flocculants and the bioflocculant, biosynthesized Fe nanoparticles possess significant potential for eliminating both biological oxygen demand (BOD) and chemical oxygen demand (COD) from wastewater samples treated. Hence, the Fe nanoparticles synthesized in this way have the potential to substitute chemical flocculants in the treatment of wastewater.

Gingko biloba L.’nın Fitokimyasal Bileşimi, Antioksidan ve Anti-Kanser Aktiviteleri Üzerine Bir Araştırma

The study aims to investigate the phytochemical composition, antioxidant potential, and anti-cancer activities of Ginkgo biloba L. leaf methanol extract. In this study, the phytochemical compounds, total phenolic and flavonoid contents of G. biloba leaf methanol extract were investigated. Additionally, antioxidant and anti-cancer activities [against HT-29 (human colon cancer line), HeLa (human cervical cancer line), and HEK-293 (human embryonic kidney cell line)] were assayed. The main phytochemical compounds were identified as gallic (0.94±0.01 mg/g), p-hydroxy benzoic (0.71±0.01 mg/g), and protocatechuic (0.60±0.02 mg/g) acids in G. biloba leaf methanol extract by HPLC. The total phenolic and total flavonoid contents were measured as 71.20±0.42 µg GAE/mg extract and 13.24±0.35 µg QE/mg extract, respectively. The high antioxidant activity was found in ABTS•+ assay (89.04±0.71%) while moderate antioxidant activity was observed in DPPH• (43.31±0.75%), metal chelating (49.04±0.49%), CUPRAC (absorbance: 0.85±0.01), and phosphomolybdenum (absorbance: 1.16±0.02) assays at 400 µg/mL. The IC50 values of G. biloba leaf methanol extract on HT-29, HeLa, and HEK-293 cell lines were recorded as 406.70±1.55, 84.86±0.98, and >800 µg/mL, respectively. The present study features a new addition to the antioxidant and anti-cancer properties of the therapeutically valuable G. biloba with its phytochemical content.

Design, synthesis, and unraveling the antibacterial and antibiofilm potential of 2-azidobenzothiazoles: insights from a comprehensive in vitro study.

The present study reports the synthesis of 2-azidobenzothiazoles from substituted 2-aminobenzothiazoles using sodium nitrite and sodium azide under mild conditions. All the synthesized compounds were examined for their antibacterial activity against Gram (+) bacteria, Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 51299), Bacillus cereus (ATCC 10876) and Gram (-) bacteria, Escherichia coli (ATCC 10536), Pseudomonas aeruginosa (ATCC 10145), Klebsiella pneumonia (ATCC BAA-2146)and clinical isolates of Gram (+) Methicillin Resistant S. aureus (MRSA) and Multi Drug Resistant E. coli. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values by broth dilution method revealed that compound 2d exhibited significant antibacterial potential against E. faecalis and S. aureus with MIC of 8μg/mL, while other synthesized compounds had only moderate effects against all the tested species. The compound significantly inhibited the biofilm formation of the bacterial strains below its MIC. The selective cytotoxicity of Compound 2d towards bacterial cells was evidenced on extended exposure of Human Embryonic Kidney-293 cell line to higher concentrations of the compound. Hence, the present study confirmed that compound 2d can be a potential drug candidate for future development as an antibacterial drug.

Nitro-Substituted Pyridinimine Complexes of Pd(II): Synthesis and Inhibition of MAO-B ex vivo

The first ever synthesis of complexes [PdLCl2] (I) and [PdLBr2] (II) was successfully achieved,where L = 2,6-dimethyl-4-nitro-N-(pyridin-2-ylmethylildene)aniline, a ligand with a purported ability toinhibit monoamine oxidase B (MAO-B). To gain insight into the molecular structure of complexes Iand II, as well as the ligand precursor 2,6-dimethyl-4-nitroaniline L4 (CIF files CCDC nos. 2255106 (I),2255105 (II), 2255103 (L), 2255104 (L4)), X-ray diffraction analysis was utilized. Complex I underwent furthercharacterization to determine its stability, solubility, and lipophilicity. Cytotoxicity studies of substancesL, I, and II on human embryonic kidney cell line HEK-293 showed no evidence of cytotoxic activity. To evaluatethe inhibitory activity of new substances L, I, and II as well as established substances III−IX, selegiline,and rasagiline, ex vivo studies were conducted, establishing a structure/activity relationship.

P17-36: Ti-alloy (TiAl6Nb7) metal powder toxicity in human embryonic kidney (HEK-293T) cell line

P17-36: Ti-alloy (TiAl6Nb7) metal powder toxicity in human embryonic kidney (HEK-293T) cell line

Bio-evaluation of the 2-nitrochalcones as potential anti-lung cancer agents, inducers of apoptosis and inhibitors of protein kinase (VEGFR-2)

A series of the 2-nitrochalcones 3a–3k was synthesized and evaluated for cytotoxicity against the human lung adenocarcinoma (A549) and human embryonic kidney (HEK293-T) cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The 3-(4-fluorophenyl) 3c and the 3-(4-(1,1,2,2-tetrafluoroethoxy)phenyl derivative 3k induced early (25–29%) and late (48–60%) apoptosis of A549 cells as determined by the Annexin V-FITC/PI method. The 3-(4-fluorophenyl) 3c, 3-(4-methoxyphenyl) 3h, 3-(2,3,4-trimethoxyphenyl) 3j and the 3-(4-(1,1,2,2-tetrafluoroethoxy)phenyl derivative 3k were also found to exhibit significant inhibitory activity against vascular endothelial growth factor receptor-2 (VEGFR-2) tyrosine kinase compared to staurosporine (0.035 ± 0.002 µM) or nintedanib (IC50 = 0.021 ± 0.001 µM) with IC50 values of 31.49 ± 0.02, 39.95 ± 0.17, 36.90 ± 0.16 and 29.10 ± 0.16 µM, respectively. Molecular docking studies were also conducted on 3c and 3k as representative models to recognize the hypothetical binding motif of the title compounds within the active site of VEGFR-2.

A Comparative and Critical Analysis for In Vitro Cytotoxic Evaluation of Magneto-Crystalline Zinc Ferrite Nanoparticles Using MTT, Crystal Violet, LDH, and Apoptosis Assay.

Zinc ferrite nanoparticles (ZFO NPs) are a promising magneto-crystalline platform for nanomedicine-based cancer theranostics. ZFO NPs synthesized using co-precipitation method are characterized using different techniques. UV-visible spectroscopy exhibits absorption peaks specific for ZFO. Raman spectroscopy identifies Raman active, infrared active, and silent vibrational modes while Fourier transforms infrared spectroscopic (FTIR) spectra display IR active modes that confirm the presence of ZFO. X-ray diffraction pattern (XRD) exhibits the crystalline planes of single-phase ZFO with a face-centered cubic structure that coincides with the selected area electron diffraction pattern (SAED). The average particle size according to high-resolution transmission electron microscopy (HR-TEM) is 5.6 nm. X-ray photoelectron spectroscopy (XPS) signals confirm the chemical states of Fe, Zn, and O. A superconducting quantum interference device (SQUID) displays the magnetic response of ZFO NPs, showing a magnetic moment of 45.5 emu/gm at 70 kOe. These ZFO NPs were then employed for comparative cytotoxicity evaluation using MTT, crystal violet, and LDH assays on breast adenocarcinoma epithelial cell (MCF-7), triple-negative breast cancer lines (MDA-MB 231), and human embryonic kidney cell lines (HEK-293). Flow cytometric analysis of all the three cell lines were performed in various concentrations of ZFO NPs for automated cell counting and sorting based on live cells, cells entering in early or late apoptotic phase, as well as in the necrotic phase. This analysis confirmed that ZFO NPs are more cytotoxic towards triple-negative breast cancer cells (MDA-MB-231) as compared to breast adenocarcinoma cells (MCF-7) and normal cell lines (HEK-293), thus corroborating that ZFO can be exploited for cancer therapeutics.

Enhanced immune responses in dexamethasone immunosuppressed male rats supplemented with herbal extracts, chitosan nanoparticles, and their conjugates

Nowadays, the world is challenged with highly contagious diseases, one of their preliminary virulence mechanisms is the suppression of innate immunity. Therefore, promoting natural immunity is a good precautionary strategy. we investigated and compared the effects of several natural herbal extracts -Moringa oleifera, Ziziphus spina christi, and Saussurea costus, and chitosan nanoparticles (CS NPs)- as well as conjugated extracts with CS NPs on the immunological parameters of dexamethasone immunosuppressed (IS) male rats. The plant extracts were assessed for total flavonoids, phenolics, and antioxidant activity. The CS NPs and their conjugates were characterized using particles size, zeta potentials, and Fourier-transform infrared spectroscopy analyses. The chemical analysis of the plant extracts, CS NPs, and their conjugates was performed using energy dispersive X-ray fluorescence, and their cytotoxicity was evaluated in human lung fibroblast (WI-38) and human embryonic kidney (HEK-293) cell lines. For in vivo evaluations, 72 adult male rats were divided into 9 groups: control, IS, three plant extracts, CS, and conjugates of the three plant extracts and CS NPs. Oral supplementation (day after day) lasted for 28 days. Liver, kidney, and spleen tissue samples were collected for histopathology and Ki-67 expression analyses. The results revealed that the plant extracts and CS improved the total leukocyte counts, complement 3, complement 4, interferon-gamma, and tumor necrosis factor levels at day 28. However, the plant extract-CS NPs conjugates faster and have higher immunostimulatory effects at day 14. Furthermore, the atrophied white pulp of the spleen induced by dexamethasone was alleviated, and Ki-67 expression was elevated in all the treated groups. Conclusively, the conjugates of Moringa oleifera, Ziziphus spina christi, and Saussurea costus extract with CS NPs demonstrated more potent and rapid immune responses at lower doses and concentrations compared to the plant extracts or CS NPs alone, without causing liver or kidney injuries. Thus, supplementation of these conjugated plant extracts at lower doses and concentrations is recommended to improve immunity while considering safety considerations.

MLK3 localizes mainly to the cytoplasm and promotes oxidative stress injury via a positive feedback loop.

Activation of mixed lineage kinase 3 (MLK3) by phosphorylation at Thr277/Ser281 stimulates downstream apoptotic pathways and ultimately leads to cell injury. MLK3 is reported to localize to both the cytoplasm and nucleus in human ovarian cancer cells and immortalized ovarian epithelial cells (T80 and T90 cells), and phosphorylation at Thr477 is required for the cytoplasmic retention of MLK3 in T80 cells. However, the subcellular distribution of MLK3 in other cell types has rarely been reported, and whether phosphorylation of MLK3 at Thr277/Ser281 affects its subcellular distribution is unknown. Here, our bioinformatics analysis predicted that MLK3 was mainly distributed in the cytoplasm and nucleus. In the human HEK293T embryonic kidney cell line and murine HT22 hippocampal neuronal cell line, endogenous MLK3 was more abundant in the cytoplasm and less abundant in the nucleus. In addition, overexpressed Myc-tagged MLK3 and EGFP-tagged MLK3 were also observed to localize mainly to the cytoplasm. MLK3 that was activated by phosphorylation at Thr277/Ser281 was mainly distributed in the cytoplasm, and phosphorylation deficient (T277A/S281A) and mimic (T277E/S281E) mutants both showed distributions similar to that of wild type (wt) MLK3, further proving that phosphorylation at Thr277/Ser281 was not involved in regulating MLK3 subcellular localization. In HEK293T cells, H2O2 stimulation accelerated MLK3 phosphorylation (activation), and this phosphorylation was reduced by the antioxidant N-acetylcysteine in a dose-dependent manner. Overexpressing wt MLK3 promoted the production of intracellular reactive oxygen species and increased cell apoptosis, both of which were enhanced by the phosphorylation-mimic (T277E/S281E) MLK3 variant but not by the phosphorylation-deficient (T277A/S281A) MLK3 variant. These findings provided additional evidence for the cytoplasmic and nuclear distribution of MLK3 in HEK293T cells or HT22 cells and revealed the pivotal role of MLK3 in the positive feedback loop of oxidative stress injury.

Cold and warmth intensify pain-linked sodium channel gating effects and persistent currents.

Voltage-gated sodium channels (Nav) are key players in excitable tissues with the capability to generate and propagate action potentials. Mutations in the genes encoding Navs can lead to severe inherited diseases, and some of these so-called channelopathies show temperature-sensitive phenotypes, for example, paramyotonia congenita, Brugada syndrome, febrile seizure syndromes, and inherited pain syndromes like erythromelalgia (IEM) and paroxysmal extreme pain disorder (PEPD). Nevertheless, most investigations of mutation-induced gating effects have been conducted at room temperature, and thus the role of cooling or warming in channelopathies remains poorly understood. Here, we investigated the temperature sensitivity of four Nav subtypes: Nav1.3, Nav1.5, Nav1.6, and Nav1.7, and two mutations in Nav1.7 causing IEM (Nav1.7/L823R) and PEPD (Nav1.7/I1461T) expressed in cells of the human embryonic kidney cell line using an automated patch clamp system. Our experiments at 15°C, 25°C, and 35°C revealed a shift of the voltage dependence of activation to more hyperpolarized potentials with increasing temperature for all investigated subtypes. Nav1.3 exhibited strongly slowed inactivation kinetics compared with the other subtypes that resulted in enhanced persistent current, especially at 15°C, indicating a possible role in cold-induced hyperexcitability. Impaired fast inactivation of Nav1.7/I1461T was significantly enhanced by a cooling temperature of 15°C. The subtype-specific modulation as well as the intensified mutation-induced gating changes stress the importance to consider temperature as a regulator for channel gating and its impact on cellular excitability as well as disease phenotypes.

Green solvents-based rutin extraction from Sophora japonica L.

Using green solvents to extract bioactive ingredients from medicinal herbs is a new trend for sustainable development procedures. The main objective of this study was to evaluate the ability to extract rutin from Sophora japonica L. using deep eutectic solvents (DESs), recover the target compound from extract, and reuse the solvent. Additionally, the antioxidant activity and cytotoxicity of the DESs extracts were also assessed. DESs of choline chloride and ethylene glycol (ChCl-Eth) showed the highest extraction efficiency of rutin from Sophora japonica L. The main factors affecting extraction yields were optimized by the response surface methodology (RSM). Under optimized conditions, the extraction yields using DESs and methanol as solvents were 26.20 % and 19.49 %, respectively. Water (as an antisolvent) was used to recover rutin from the DESs extract with a recovery yield of 94.9 %. The DESs solvent was recovered and recycled at least three times without loss of extraction yields. The DESs of ChCl-Eth had no significant effect on the antioxidant activity of rutin and the recovered rutin has exhibited more antioxidant activity than methanol extract. The selected greens solvents and their extracts did not show cytotoxic activity against the human embryonic kidney cell line (HEK-293A). The results serve as a basis for the development of other studies, as a premise to deploy a safe and environmentally friendly process of extracting natural active ingredients on an industrial scale.