Normal Kidney Epithelial Cells Research Articles

#2695 A Polycystin-1-Bicaudal C1 complex regulates focal adhesion function through integrin beta-1, F-actin organisation and local translation in HKE cells

Abstract Background and Aims Variants in Bicc1, result in a recessive murine PKD phenotype (bpk, jcpk) by uncertain mechanisms. In a recent study, we reported genetic and biochemical evidence of novel interactions between PKD1, PKD2 and BICC1 in Xenopus, mouse and man suggesting that BICC1 is a modifier of the ADPKD phenotype (submitted). In this study, our major aim was to investigate the molecular mechanisms underlying cystogenesis that result from loss of the normal interaction between the PKD1 protein, Polycystin-1 (PC1), a cell surface receptor and Bicaudal C1 (BICC1), an RNA-binding protein. Methods Isogenic PKD1 and BICC1 CRISP-Cas9 KO cells were generated from a conditionally-immortalised wild-type human proximal tubular line (UCL93); RNA-seq was performed in PKD1 and BICC1 KO cells to identify commonly altered genes and pathways; MS pull-down was conducted in HEK293 cells with BICC1-HA to identify BICC1 interacting proteins; cellular phenotypes in normal and KO cells were assessed through adhesion and migration assays, alongside Traction Force Microscopy (TFM); proximal ligation assays (PLA) to confirm protein-protein interactions; single-molecule FISH (smFISH) to localize mRNA transcripts; puromycin labeling with a proximity ligation assay (Puro-PLA) to detect sites of translation of specific mRNAs. Results Consistent with a functional interaction, loss of PC1 resulted in lower BICC1 levels and vice-versa in single KO (SKO) cells. In migrating cells, BICC1 was expressed at the leading edge where it co-localised with β1-integrin (ITGB1) by PLA, suggesting enrichment at focal adhesion sites (FA). BICC1 KO cells had no cilia phenotype but instead displayed striking disorganization of stress fibres and aster-like structures, features which could be completely rescued by lentiviral Bicc1-HA re-expression. Gene set enrichment and pathway analysis confirmed that F-actin organization and actin-associated proteins (AAP) were commonly dysregulated in SKO cells. BICC1 was also found to interact ITGB1, several components of an integrin-associated complex and proteins involved in actin turnover by MS pull-down. Both SKO cells showed major abnormalities of cell attachment, detachment and directional migration indicative of functional alterations in cell-matrix and cell-cell adhesion. We confirmed that FA structure and function were altered by reduced co-localisation of several AAPs (including TNS1 and FBLIM1) and by TFM. BICC1 directly interacted with F-actin and FBLIM1 in pull-down assays and with FBLIM1 by RNA-IP. Finally, we found evidence of FBLIM1 mRNA by smFISH and of local FBLIM1 translation (Puro-PLA) at FA sites in wild-type cells, both of which were markedly reduced in SKO cells. Conclusion Our data reveal the key roles for actin dynamics and local translation as novel mechanisms regulated by PC1 and BICC1 in the pathogenesis of ADPKD. We demonstrate that BICC1 and PC1 are co-regulated, forming a PC1-BICC1 complex critical for maintaining the structural integrity and function of FAs in normal kidney epithelial cells. Loss of this interaction results in major alterations in cell adhesion, migration and stress fibre organisation, features typical of the ADPKD cellular phenotype. BICC1 plays a dual role in this process via protein-protein interactions with ITGB1, F-actin and other AAPs, and through protein-RNA interactions regulating the local translation of key AAPs (FBLIM1) at FA sites.

Enhanced stability and dissolution of curcumin nanocrystals stabilized by octenyl succinic anhydride modified starch

Curcumin (CUR) has known to have poor oral bioavailability due to its poor aqueous solubility and stability in gastrointestinal fluid condition as well as its limited permeability which greatly limits its applications in food and medical fields. The present study reported the development of CUR nanocrystal suspension (CUR-NS) for enhanced bioavailability, stabilized with octenyl succinic anhydride modified starch (OSA-S), a naturally renewable and biocompatible biomolecule. This preparation was achieved via an antisolvent precipitation combined with ultrasonic dispersion method, following which the suspension was freeze-dried to form powder CUR nanocrystals (CUR-NCs). Physicochemical properties of the prepared CUR-NS and CUR-NCs powders were extensively characterized. The particle size of the nanocrystals was found to be around 322.8 ± 15.7 nm with a small polydispersity index (PDI) of about 0.230 ± 0.039. Rod like crystal shape was observed by the SEM. Results from powder X-ray diffraction (pXRD) and differential scanning calorimetry (DSC) indicated that the crystal form of CUR in CUR-NCs was a monoclinic polymorph Form 2. Stability results indicated that CUR-NCs possessed an excellent storage-, thermal-, and photo-stabilities. CUR-NCs had a significantly higher dissolution rate, a 68-fold increase in its solubility than that of raw CUR. The cytotoxicity assay demonstrated that CUR-NCs exhibited lower toxicity towards normal human kidney epithelial cells compared to raw CURs. Parallel artificial membrane permeability assay (PAMPA) characterization confirmed that the flux of CUR was significantly enhanced after forming nanocrystals with OSA-S, probably resulting from its increased solubility and dissolution rate. The study showed that OSA-S could effectively stabilize CUR-NCs, and CUR-NCs developed in the current work exhibited significant enhancement in the stability and dissolution rate leading to an improved bioavailability which might be helpful for further development of CUR based products.

Eu3+ doped SrMoO4: A promising nanophosphor for enhanced fingerprint detection and cheiloscopy

Developing a single material with diverse applications like fingerprint detection, cheiloscopy, and anti-counterfeiting is truly innovative and has the potential to revolutionize various fields. Here, we synthesized nano phosphors powders of SrMoO4 nanoparticles, doped with spherical-shaped Eu3+ ions via co-precipitation method. X-ray diffraction (XRD) analysis confirmed the formation of the SrMoO4 phase with Eu3+ successfully incorporated into the lattice. Field emission scanning electron microscopy (FESEM) images revealed the nanoparticles to be spherical and monodispersed with a narrow size distribution. Energy dispersive X-ray spectroscopy (EDS) confirmed the presence of Sr, Mo, Eu3+, and O elements, indicating a homogeneous elemental composition. Fourier transform infrared (FTIR) spectroscopy showed characteristic bands for MoO bonds and hydroxyl groups. Diffuse reflectance spectroscopy (DRS) analysis revealed a decrease in band gap with increasing Eu3+ concentration, suggesting that Eu3+ doping potentially modifying the optical properties of SrMoO4, and introducing new functionalities. Their red photoluminescence property, making them suitable for various optical applications. The nanoparticles showed potential applications in fingerprint detection, cheiloscopy (lip print analysis), and anti-counterfeiting due to their red luminescence, small size, and biocompatibility. The developed fingerprints showed excellent contrast, high sensitivity and selectivity on various substrates. In vitro cytotoxicity studies indicated that the nanoparticles were non-toxic to normal kidney epithelial cells at the tested concentrations. Thess versatilities demonstrate the potential of engineered materials for criminal identification in forensic applications. Therefore, they play a crucial role in solving crimes and bringing justice to victims.

The Apoptotic Property of Nymphaea Caerulea Flower Extract on Acute Myeloid Leukaemia Cell Line, THP-1.

Acute Myeloid Leukaemia (AML) is considered to be an extremely heterogeneous malignancy of bone marrow and blood. The first line of therapy for AML is prolonged chemotherapy. Due to the presence of molecular heterogeneity in AML as confirmed by next-generation sequencing, researchers are planning to develop newer strategies of therapy. In the present study we have explored the anti-cancer potentiality of the hydro-ethanolic extract (50% and 70%) of the whole flower of Nymphaea caerulea against the Acute Myeloid Leukaemia cell line, THP-1 with control of normal human kidney epithelial cell line (HEK 293). The present study is a novel contribution to the existing scientific knowledge as at present no study as an anti-leukaemic agent is available on N. caerulea (blue lotus) extract and exploring its action mechanism on in-vitro cell line model. Some targeted cytokine and apoptotic genes genes to deduce the anti-cancer mechanism of action of the crude extract (hydro-ethanolic extract (50% and 70%) of the whole flower) were selected as Interferon (IFN) γ, Interleukins - IL-6, IL-8, IL- 10, IL-1β, Transforming Growth Factor (TGF β1), Tumor Necrosis Factor (TNF α), Caspase 3(CAS 3), Caspase 9 (CAS 9), CD95 (Fas), Tumor Necrosis Factor Receptor 1 (TNFRSF1A) to observe relative fold changes of the expression using Real-Time PCR with housekeeping gene β-actin. Cellular cytopathic effect (CPE), cell viability assay by methylene blue assay, and cell cytotoxicity of the crude extract against the THP-1 cell line were also studied along with it's bio-active compositional analysis of the extract was explored using ultra-performance liquid chromatography followed by mass spectra. The N. caerulea flower extract is capable of inducing apoptosis in AML and it can balance cytokine alterations in such diseases. Nymphaea caerulea flower extract appears to be a good anti-leukemia agent.

Deficiency of ligase IV leads to reduced NHEJ, accumulation of DNA damage, and can sensitize cells to cancer therapeutics

Ligase IV is a key enzyme involved during DNA double-strand breaks (DSBs) repair through nonhomologous end joining (NHEJ). However, in contrast to Ligase IV deficient mouse cells, which are embryonic lethal, Ligase IV deficient human cells, including pre-B cells, are viable. Using CRISPR-Cas9 mediated genome editing, we have generated six different LIG4 mutants in cervical cancer and normal kidney epithelial cell lines. While the LIG4 mutant cells showed a significant reduction in NHEJ, joining mediated through microhomology-mediated end joining (MMEJ) and homologous recombination (HR) were significantly high. The reduced NHEJ joining activity was restored by adding purified Ligase IV/XRCC4. Accumulation of DSBs and reduced cell viability were observed in LIG4 mutant cells. LIG4 mutant cells exhibited enhanced sensitivity towards DSB-inducing agents such as ionizing radiation (IR) and etoposide. More importantly, the LIG4 mutant of cervical cancer cells showed increased sensitivity towards FDA approved drugs such as Carboplatin, Cisplatin, Paclitaxel, Doxorubicin, and Bleomycin used for cervical cancer treatment. These drugs, in combination with IR showed enhanced cancer cell death in the background of LIG4 gene mutation. Thus, our study reveals that mutation in LIG4 results in compromised NHEJ, leading to sensitization of cervical cancer cells towards currently used cancer therapeutics.

Characterization of the Cellular Origin and Oncogenic Mechanisms of Chromophobe Renal Cell Carcinoma (ChRCC) and Renal Oncocytic Neoplasms

Abstract Background ChRCC is a rare form of kidney cancer and presents with a poor prognosis in the metastatic setting, with limited response to immune checkpoint inhibitors (ICI) and targeted therapy. While previous studies suggested that ChRCC originates from intercalated cells of the kidney, its exact cellular origin has not been clearly defined. We therefore investigated the cellular origin of ChRCC and renal oncocytic neoplasms at single-cell resolution. Methods ChRCC, renal oncocytoma (RO) and low-grade oncocytic tumor (LOT) samples with matched normal kidney specimens were analyzed using single-cell RNA sequencing (scRNA-seq). Epithelial cells from normal samples were clustered and annotated into the distinct known cellular types found in the adult healthy human kidney. A logistic regression model (Young M.D. et al., Science, 2018) was trained on the normal epithelial clusters, using a comprehensive set of 74 marker genes (features). Subsequently, the model was tested on tumor clusters from ChRCC, RO and LOT to investigate their cellular origin through the identification of the highest predicted probabilities of similarity between normal epithelial cellular types and tumor cells. Differential gene expression and pathway analyses between ChRCC and its cell of origin were then conducted to investigate mechanisms of oncogenesis. Results Following quality-control, 46,817 cells from 5 tumors (ChRCC: n=3, RO: n=1 and LOT: n=1) and 4 normal samples were isolated for scRNA-seq analysis. Among normal samples, 784 epithelial cells were clustered and annotated into the following cellular entities: proximal tubule (PT), loop of Henle – distal tubule (LOH-DT), principal cells (PC), α-intercalated cells (ICA) and β-intercalated cells (ICB). Across all ChRCC and oncocytic tumors, 7,573 tumor cells were identified, among which 7,425 corresponded to ChRCC. For ChRCC, RO, and LOT neoplasms, the highest predicted probability of similarity was consistently identified with ICA cells (ranging from 0.60 to 0.80). This finding was validated using an external and previously published (Zhang Y. et al., Proc Natl Acad Sci USA, 2021) scRNA-seq dataset from a patient with ChRCC (n=2,853 cells). Additionally, evaluation of previously defined ChRCC-specific markers (i.e. FOXI1, RHCG, KIT, CLCNKA, and CLCNKB) in scRNA-seq data of normal kidney epithelial cells consistently showed the highest expression in ICA cells. Differential gene expression between ChRCC and ICA cells revealed a significant downregulation of MHC class I genes (i.e. HLA-A , HLA-B, and HLA-C) and HSP70 (family A) genes (i.e. HSPA1A, HSPA1B, HSPA6, and HSPA8) in ChRCC. Differential pathway analysis between ChRCC and ICA cells showed a significant enrichment of the ferroptosis, glutathione metabolism, mTOR signaling and IL-15 pathways in ChRCC. Conclusions Renal oncocytic tumors, including ChRCC, appear to originate from the α-intercalated cells of the normal human kidney. As compared to α-intercalated cells, ChRCC downregulates MHC class I genes, which may be associated with its poor response to immunotherapy. Further, the selective downregulation of HSP70 genes may in part explain ChRCC’s increased sensitivity to ferroptosis. CDMRP DOD Funding: yes

N-(2-hydroxy phenyl) acetamide ameliorate inflammation and doxorubicin-induced nephrotoxicity in rats

Doxorubicin (DOX) is an anthracyclin antibiotic used for the treatment of various cancers. Nephrotoxicity is among the serious side effects of DOX, therefore, DOX-induced nephrotoxic model has been widely used to study nephropathies. The objectives of this study is to investigate the possible anti-inflammatory and nephroprotective effects of salicylic acid derivative, N-(2-hydroxy phenyl) acetamide (NA-2), in a rat model of DOX-induced nephrotoxicity. The in vitro anti-inflammatory potential of NA-2 was manifested by whole blood oxidative burst and nitric oxide (NO) assays with no toxicity on normal human fibroblast (BJ) cells, human embryonic kidney (HEK-293) cells, and normal monkey kidney epithelial (Vero) cells. The in vivo study included five groups: Normal control, DOX (6 mg/kg DOX-i.v.via tail vein), NA-2 treated control-i.p., NA-2/DOX treated-i.p., and prednisolone/DOX treated. After 7 days of DOX administration, rats with urinary protein level of >50 mg/kg/day were selected. Treatment group rats received i.p. doses of NA-2 (10 mg/kg/day) for 3 weeks with weekly monitoring of urinary protein excretion and body weights. mRNA expression of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP)-1, and kidney injury molecule (KIM)-1 was analyzed by quantitative polymerase chain reaction (qPCR). Protein expressions were analyzed by immunohistochemistry. NA-2 attenuated DOX-induced changes in serum and urine levels, and improved inflammatory profile of the renal tissue. Histopathological findings revealed protective effects of NA-2 showing lesser lesions. We conclude that NA-2 is able to protect against DOX-induced renal damage functionally, biochemically and histopathologically with corresponding improvement in the kidney inflammatory profile.

Double advantages of 2D coordination polymer of coumarinyl‐pyridyl Schiff base decorated Zn(II): The fabrication of Schottky device and Anti‐carcinogenic activity

A 2D coordination polymer, [Zn2(BDC)4(QPR)2(H2O)]n (Zn(II)‐CP) (BDC2− = 1,4‐benzenedicarboxylato; QPR = 7‐[(pyridin‐4‐ylmethylene)‐amino]‐chromen‐2‐one), is presumably the first reported on coumarinyl‐pyridyl framework with terephthalato linker. The optical band gap, 2.91 eV, is obtained from the Tauc's plot that insists to fabricate an optically stimulated semiconducting device, and the electrical conductivity exhibited (Λ) is 8.07 × 10−3 S m−1 (dark) and 9.26 × 10−3 S m−1 (light). The cytotoxicity in the selected human cancer cell lines like HeLa, MDA‐MB 231, and HepG2 cells in relative to the normal kidney epithelial (NKE) cell line has been examined, and its efficacy is observed highest in inhibiting proliferation of HeLa cells (IC50, 21.02) compared with other cancer cells (IC50: 42.22 [HepG2] and 46.37 [MDA‐MB 231]). Further, the Zn(II)‐CP possess intracellular reactive oxygen species (ROS) generation and caspase‐3/7 activation leading to G2/M cell cycle arrest and apoptotic cell death in HeLa cell line. This novel coumarinyl‐pyridyl ligand appended Zn(II)‐CP has been employed for the fabrication of semiconducting device, which is quite impressive in view of existing energy crisis civilization; in addition, the compound has excellent bio‐activity.

Cytoprotective Properties of Citronella Oil (Cymbopogon nardus (L.) Rendl.) and Lemongrass Oil (Cymbopogon citratus (DC.) Stapf) through Attenuation of Senescent-Induced Chemotherapeutic Agent Doxorubicin on Vero and NIH-3T3 Cells.

This study aimed to determine the cytoprotective potentials of citronella (Cymbopogon nardus (L.) Rendl.) essential oil (CO) and lemongrass (Cymbopogon citratus (DC.) Stapf) essential oil (LO). The essential oils from citronella and lemongrass were obtained by steam-water distillation, then analyzed using Gas Chromatography-Mass Spectrophotometry (GC-MS) to determine the chemical constituents. The antioxidant activity of CO and LO was compared using a total antioxidant capacity kit. The viability of normal kidney epithelial cells Vero and fibroblast NIH-3T3 as the cell models were tested using a trypan blue exclusion assay. The effect of cellular senescence inhibition on both cell models was measured using senescence-associated β-galactosidase (SA-β-gal) staining. The mechanism of action of CO and LO in the protection of cellular damage against doxorubicin was also confirmed through 2',7'-dichlorofluorescin diacetate (DCFDA) staining to discover the ability to decrease reactive oxygen species (ROS) levels and a gelatin zymography assay to observe the activity of matrix metalloproteinases (MMPs). The major marker components of CO and LO were citronellal and citral, respectively. Both oils showed low cytotoxic activity against Vero and NIH-3T3 cells, with IC50 values of over 40 µg/mL. LO exhibited higher antioxidant capacity than CO, but there was no effect on the intracellular ROS level of both oils on Vero and NIH-3T3 cells. However, CO and LO decreased cellular senescence induced by doxorubicin exposure on both cells, as well as suppressed MMP-2 expression. Conclusion: Both CO and LO decrease the cellular senescence and MMP-2 expression with less cytotoxic effects on normal cells independently from their antioxidant capacities. The results were expected to support the use of CO and LO as tissue protective and anti-aging agents in maintaining the body's cellular health against chemotherapeutics or cellular damaging agents.

Potent saccharinate-containing palladium(II) complexes for sensitization to cancer therapy

Palladium(II) (PdII) complexes are among the most promising anticancer compounds. Both 2`-benzoylpyridine thiosemicarbazone (BpT) and saccharinate (Sac) are efficient metal chelators with potent anticancer activity. To explore a more effective new anticancer drug, we synthesized a series of Sac and BpT-containing PdII complexes coordinated with thiosemicarbazone (TSC)-derived ligands, and characterized them through nuclear magnetic resonance (NMR), Fourier transformed infrared spectroscopy (FT-IR), elemental analysis, ultraviolet-visible spectroscopy (UV–Vis) and thermogravimetric analysis (TGA). Each target complex was composed of PdII, BpT, and one or two Sac molecules. Both the in vitro and in vivo anti-growth effects of those ligands and the obtained PdII complexes were investigated in the human lung adenocarcinoma cell lines A549 and Spc-A1. The coordination of PdII with the TSC-derivatives and Sac resulted in clearly greater anticancer activity than single ligands. These compounds were demonstrated to be safe for 293 T normal human kidney epithelial cells. The introduction of Sac into the TSC-derived PdII complex significantly enhanced anti-growth effects, and induced apoptosis of human lung cancer cells in vitro and in vivo in a dose dependent manner. Moreover, the PdII complex containing two Sac molecules showed the most promising therapeutic effects, thereby confirming that Sac increases the cancer therapeutic efficacy of PdII complexes and providing a new strategy for exploring anticancer drugs for potential clinical treatment.

Anticancer and Neuroprotective Effects of the Triterpene Glycosides From Sea Cucumber Holothuria imitans

Introduction: Sea cucumbers has gained notoriety because possess a wide range of biological and pharmacological activities. In this sense, the aim of this work was to evaluate the anticancer and neuroprotective effects of the triterpene glycosides from sea cucumber Holothuria imitans. Methods: Triterpene glycosides were separated and purified by Reversed-phase high-performance liquid chromatography (RP-HPLC). Their structures were deduced by spectral analysis and chemical evidence. Cytotoxic activity was evaluated using normal African green monkey kidney epithelial cell line (VERO) and three cancer cell lines: cancer gastric (MKN-28), breast adenocarcinoma (MCF-7) and lung carcinoma (A-549). Besides, the neuroprotective effect was studied using the Cath.a-differentiated (CAD) cell line and human glial (Oligodendrocytic) hybrid cell line (MO3.13). Results: Two triterpene glycosides (Fuscocineroside C and Scabraside D) were isolated, which showed low cytotoxic activity against VERO cell line, and high cytotoxic activity against lines MKN-28, MCF-7 and A-549 cells, with IC50 between the ranges of 0.92 μmol/L to 2.61 μmol/L. The isolated triterpene glycosides showed the ability to regain mitochondrial viability in CAD and MO3.13 cells treated with neurotoxin (C2-ceramide) with statistically significant results (p<0.05). Conclusion: The triterpene glycosides Fuscocineroside C and Scabraside D isolated from sea cucumber Holothuria imitans show anticancer and neuroprotective potential and may be considered promising active principles for anticancer and neuroprotective drugs.

Synthesis, structural characterization and in vitro cytotoxicity assessment of new mononuclear Cu(II) and Co(II) complexes against MDA–MB–231, HCC–1806 and HT–29 cancer cell lines

In the present work, we report the synthesis, structural characterization and in vitro biological evaluation of four mononuclear Cu(II) and Co(II) complexes 1–4 derived from the Schiff base ligand scaffolds L1 and L2. The synthesized complexes 1–4 were systematically characterized by various analytical and spectroscopic techniques (FTIR, EPR, UV–vis and sXRD). DFT calculations were performed to determine the structural, electronic and chemical properties of complexes 1–4 based on their respective HOMO and LUMO energy values. Hirshfeld surface analysis was carried out to examine the intermolecular interactions (CH···O, NH···H, Cl–H···H and OH···O) which define the stability of the crystal lattice. Furthermore, comparative in vitro DNA/BSA binding studies were carried out by employing various biophysical methods which revealed a higher binding propensity for the copper analogues. The antioxidant activity of complexes 1–4 was also evaluated against the free radical DPPH and the results demonstrated a better radical scavenging activity by the copper analogues. In addition, the cytotoxicity of the complexes was evaluated using an SRB assay against MDA–MB–231/HCC1806 (triple negative breast cancer cell lines) and HT–29 (colorectal cancer cell lines), which revealed remarkable cytotoxicity of complex 3 against the tested cancer cell lines. The toxicity profile of the complexes was also evaluated against VERO (normal kidney epithelial) cells, which validated their low toxicity behaviour.

Synthesis, spectroscopic characterizations, single crystal X-ray analysis, DFT calculations, in vitro biological evaluation and in silico evaluation studies of thiosemicarbazones based 1,3,4-thiadiazoles

We created new thiosemicarbazones and their cyclized heterocyclic compounds, examined them, and evaluated their biological activity in silico and in vitro. Two pyrene-based thiosemicarbazone compounds, (E)-N'-(pyren-1-ylmethylene) pyrrolidine-1-carbothiohydrazide (PP1) and (E)-N'-(pyren-1-ylmethylene) morpholine-4-carbothiohydrazide (PM1) were prepared from pyrrolidine-1-carbothiohydrazide and morpholine-4-carbothiohydrazide with pyrene-1-carboxaldehyde. The 2-(pyren-1-yl)-5-(pyrrolidin-1-yl)-1,3,4-thiadiazole (PP2) and 4-(5-(pyren-4-yl)-1,3,4-thiadiazol-2-yl)morpholine (PM2) were obtained unexpectedly from the cyclization of PP1 and PM1 in the presence of Mn(OCOCH3)2⋅4H2O. FT-IR, UV-Visible, NMR, and HR-MS were used to accurately portray the structure of the compounds. The molecular composition of PP1, PP2 and PM2 were ascertained by the single-crystal X-ray diffraction (XRD) method. The compounds are ordered and dispersed in three dimensions according to the density functional theory (DFT) findings, which are in good agreement with the XRD-derived crystal structures. The molecular docking analysis revealed the various possible interactions of drugs with the non-small cell lung cancer protein (EGFR with a non-covalent inhibitor). The binding energies for the compounds were found to be in the following order PP2 > PP1 > PM1 > PM2, and molecular dynamic simulation was used to further examine the binding mechanism of the most active compound (PP2). These compounds bound strongly with calf thymus (CT)-DNA, which was well described by absorption and emission studies. This demonstrated that PP2 had a better ability to bind (Kb value of 1.95 × 106 and Kq value of 2.43 × 104). Similarly, the synergy of bovine serum albumin (BSA) with compounds also was examined and showed PP2's significant binding potential (Kb value of 2.94 × 104 and Kq value of 6.87 × 104). Additionally, using an MTT assay against a few human cancer cells, the substances' in vitro cell viability was evaluated, including HepG-2 (hepatocellular carcinoma) and T24 (bladder), as well as Vero (normal kidney epithelial cells isolated from an African green monkey). With IC50 values of 13.16 μM (Cisplatin-IC50 = 49.9 μM) and 37.67 μM (Cisplatin-IC50 = >50 μM), respectively, the findings demonstrated that PP2 displayed efficient cytotoxicity towards HepG-2 and T24 cells.

Pivotal Role of Copper Nanoparticles Shelled by Turmeric or Sumac on Huh-7 Cell Line Cytotoxicity, Apoptosis and Antioxidant Capacity

&lt;b&gt;Background and Objective:&lt;/b&gt; Cancer is a complex interaction among multiple signalling pathways involving a variety of target molecules. Nanoparticles were used in cancer treatment because of their intrinsic anticancer properties. The use of plant extracts in the preparation of metallic nanoparticles as a convenient substitute has been proposed. This study assessed the cytotoxic, antioxidant and apoptotic effects of copper nanoparticles shelled with either turmeric or sumac biosynthesized as core-shell nanostructures on the liver tumour cell line (Huh-7). &lt;b&gt;Materials and Methods:&lt;/b&gt; The nanostructures were synthesized by sonochemical method and characterization was done to confirm the successful synthesis within the nanoscale. Cytotoxicity of nanostructures was investigated on Huh-7 and normal kidney epithelial cell lines (VERO). Malondialdehyde, nitric oxide, reduced glutathione and superoxide dismutase were estimated in cell lysate to assess the antioxidant properties of nanostructures. Caspase-3 was also measured as an apoptotic marker. &lt;b&gt;Results:&lt;/b&gt; Both nanostructures had low IC&lt;sub&gt;50&lt;/sub&gt; on Huh-7 cells and a non-toxic effect on VERO cells. The cytotoxic effect was coupled with a significant increase in antioxidant activities and apoptotic efficiency compared to control. &lt;b&gt;Conclusion:&lt;/b&gt; The findings summarized here support the utilization of biosynthesized copper with turmeric or sumac as core-shell nanostructures as a novel chemotherapeutic drug for cancer treatment that improves antioxidant effect that modulates the side effect of cytotoxicity. Also, it is obvious that copper nanostructure biosynthesized with turmeric has a more advanced effect than that of sumac.

Polymer nanoparticle delivery of microRNA-1303 suppresses renal cell carcinoma differentiation via regulation of small nucleolar RNA host gene 16

Small nucleolar RNA host gene 16 (SNHG16) participates in some cancers as microRNA (miR)-1303 is also reported to function on proliferation of various cancer cells. This study aimed to assess SNHG16 and miR-1303′s role in renal cell carcinoma (RCC) and its underlying mechanism. RT-qPCR was used to determine SNHG16 and miR-1303 expression in RCC cells (A498, 786-O, ACHN and OS-RC-2) and normal kidney epithelial cells (HK-2). Functional experiment was established to detect the role of miR-1303. After synthesis of nanoparticles carrying miR-1303 and transfection, CCK-8 method and assays were used to evaluate cell growth and apoptosis. The target genes for miR-1303 were predicted using bioinformatics software, and binding of miR-1303 to SNHG16 was evaluated using the dual-luciferase reporter assay. miR-1303 was lowly expressed in RCC cells, with lowest level in A498 (p &lt; 0.05). Overexpressed miR-1303 significantly reduced proliferation ability of RCC cells and induced apoptosis (p &lt; 0.05). Besides, transfection of NP carrying miR-1303 mimic resulted in dramatically decreased migrated cells when reducing the expression of SNHG18 mRNA. The presence of NPs strengthened the inhibitory effect of miR-1303 on RCC. Furthermore, the miR-1303+SNHG16-WT co-transfection group had lower relative luciferase activity compared with miR-1303+SNHG16-MUT co-transfection group (p &lt; 0.05). miR-1303 was down-regulated in RCC and NP delivery of miR-1303 inhibited RCC cell proliferation and differentiation through regulation of SNHG16. These findings suggest miR-1303 may become a potential molecular target for RCC.

Synthesis of Rutin loaded nanomagnesia as a smart nanoformulation with significant antibacterial and antioxidant properties

Nowadays synthesis of hybrid nanomaterials is of interest to researchers due to their profound antioxidant and antibacterial properties. Here, at first we prepared amine functionalized positively charged MgO nanoparticles (MgO-NH2) via microwave assisted heating followed by annealing at 400 °C for 5 h, and conjugated thereafter with a natural flavonoid, Rutin to form a bioactive nanohybrid, MgO-NH2-Rutin. The nearly spherically shaped nanohybrids with size ∼35–40 nm showed drug loading content of ∼14 % with a sustained release pattern at pH 7.4. The nanohybrids exhibited superior antioxidant properties in cell free system (DPPH, hydrogen peroxide and superoxide) as well as in intracellular condition, and showed very little cytotoxicity upto a concentration of 100 μg/mL in normal kidney epithelial cells. Besides, MgO-NH2-Rutin also triggered a significant growth inhibition against both gram positive (S. aureus) & gram negative (E. coli) bacteria. Mechanistic studies showed that the nanohybrid caused bacterial cell death via oxidative stress, DNA degradation and cell membrane disruption. Therefore, our research highlighted the use of MgO nanoparticles for the synthesis of MgO-NH2-Rutin nanohybrid, which could enhance the pharmacological actions of hydrophobic drug molecules and exhibit synergistic antioxidant and antibacterial activities.

Synthesis of carbon dots from taurine as bioimaging agent and nanohybrid with ceria for antioxidant and antibacterial applications

Here we have synthesized water soluble and biocompatible carbon dots (CDs) from taurine via thermal decomposition method. The CDs showed nearly spherical shape with diameter less than 10 nm. The CDs exhibited excitation dependent fluorescence emission and could be used for mammalian cell imaging. The CDs showed excellent DPPH and hydrogen peroxide radical scavenging activity in cell free system. Besides, the CDs also displayed significant intracellular radical scavenging activity in human normal kidney epithelial (NKE) cells. Furthermore, nanohybrids consisting of both CDs and nanoceria (CeO2) were prepared and tested for their biomedical applications. The nanohybrids showed significant antioxidant activities in both cell free and intracellular conditions. The CDs and nanohybrids possessed very little toxicity upto the concentration of 100 μg/mL when treated for 24 hours in human NKE cells. The CDs as well as nanohybrids further displayed significant bacterial growth inhibition against both gram-positive and gram-negative bacteria under dark as well as light illumination condition via the bacterial membrane damage. However, under the light illumination, the bacterial growth inhibition of CDs and nanohybrids was further enhanced due to the generation of reactive oxygen radicals and subsequent DNA degradation. A higher dose-dependent intracellular antioxidant and antibacterial activities of the nanohybrid is attributed to the synergistic effect of nanoceria and CDs. All these results clearly reflected that our synthesized CDs and their nanohybrids can be used for several biomedical applications.

The Effect of Adding Spices to Green Walnut Tinctures on Their Polyphenolic Profile, Antioxidant Capacity and Action on Renal Cells

Green walnuts—unripe fruits of Juglans regia L. are known for their many biological activities and therapeutic potential. Tinctures based on unripe walnuts (samples 1 and 2) and tinctures with the addition of spices (ginger, cloves, bay leaves, juniper fruit: samples 3–6) were tested for polyphenol profile and antioxidant capacity. The effect on a normal monkey kidney epithelial cell line (VERO) was evaluated. For monitoring the changes in cell proliferation, real-time cell analysis (xCELLigence system) was employed and cell viability was measured by the MTS test. All tinctures showed the presence of polyphenols, mainly phenolic acids, flavonoids, and naphthoquinones, and in the sample with cloves, eugenol was found. Addition of spices increased the antioxidant capacity. Tinctures show a dose-dependent cytotoxic effect. The lowest concentrations (125 µg mL−1) of both tinctures without additives (1 and 2) and with bay leaves (4) did not adversely affect (p &gt; 0.05) and even supported cell proliferation (p &lt; 0.05) in comparison to the control cells without treatment. Viability was lower in all cells except for the cells treated with tincture with cloves addition (p &gt; 0.05). It indicates the beneficial effect of cloves enrichment and supports the assumption that the appropriate dosage of tinctures is necessary to be safe for the consumers.

Synthesis of green carbon dots as bioimaging agent and drug delivery system for enhanced antioxidant and antibacterial efficacy

Now-a-days, synthesis of carbon dots (CDs) from renewable sources has attracted considerable attention due to their wide spread availability, lower cost, higher aqueous solubility and more biocompatibility. Here, we have synthesized water soluble CDs with size between 1 and 4 nm from the root extract of red Korean ginseng via simple microwave irradiation. The CDs showed excitation-dependent fluorescence emission and used as a bioimaging agent. The CDs were further loaded with a natural flavonoid, Rutin to check its efficacy as a drug delivery system. The CDs showed a drug loading content of 12.7 % and a sustained drug release behavior at physiological conditions. CDs showed excellent free radical scavenging activity both in cell free systems and intra-cellular conditions. Besides, loading with the drug further improved the intra-cellular radical scavenging activity. The CDs and the nanohybrids (Rutin-loaded CDs) displayed very little toxicity upto the concentration of 100 μg/mL in normal kidney epithelial cells. The CDs and the nanohybrids also showed a dose-dependent antibacterial effect against both gram negative as well as gram positive bacteria under light exposure as well as dark conditions due to membrane damage. However; under light exposure, the antibacterial activity of CDs and the nanohybrids was further increased due to the production of excessive reactive oxygen species (ROS) and subsequent DNA damage. Overall, our synthesized green CDs can be considered as a promising candidate for the biomedical applications due to its facile synthesis, low cytotoxicity, excellent fluorescent properties and potential as drug delivery systems.

Synthesis, structural, DNA/protein binding and cytotoxic studies of copper(I) ∝-diimine hydrazone complexes

A series of copper(I) complexes with α-diimine hydrazone ligands of the type [Cu(PPh3)2(L1-4)] (1–4) were synthesized by the reacting [Cu(CH3COO)(PPh3)2] with α-diimine ligands (L1-4) [L1 = 1, 2-bis(2-(benzothiazole-2-yl)hydrazineylidene)-1, 2-dihydro acenaphthylene (AQBH), L2 = 1, 2-bis(2-(quinolin-2-yl)hydrazineylidene)-1, 2-dihydro acenaphthylene (AQQH), L3 = 9, 10-bis(2-(benzothiazol-2-yl)dihydrazano)phenanthren-9,10-one (PQBH), L4 = 9,10-bis(2-(quinolin-2-yl)dihydrazano)phenanthren-9(10H)-one (PQQH)]. The new complexes were characterized by elemental analysis, UV–vis, FT-IR, 1H 13C NMR spectra and electrospray ionization-mass spectrometry (ESI-MS). Especially, the solid state structure of L1 (AQBH) was established using single crystal X-ray analysis. The interaction of complexes with CT-DNA was explored in detail using absorption and emission spectral methods to gain some insight into the structure–activity relationship. The obtained results revealed that complexes could interact with CT-DNA via intercalation. The interaction of these synthesized Cu (I) complexes with bovine serum albumin (BSA) was also evaluated using absorption and fluorescence techniques, which provided a static quenching mechanism between them. In addition, the cytotoxicity of compounds against HepG-2 (hepatic carcinoma) cancer and Vero normal (kidney epithelial cells extracted from an African green monkey) cells was evaluated by MTT assay. It was found that complex 4 (19.54 µM) exhibited potential activity towards HepG-2 cells which was more efficient than cisplatin (48.50 µM).