Kidney Epithelial Cell Line Research Articles

The role of efflux transporters in cytotoxicity and intracellular concentration of chlorpyrifos and chlorpyrifos oxon in human cell lines

In this study, we investigated the role of two efflux transporters, p-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), in the cytotoxicity and intracellular accumulation of the organophosphate pesticide chlorpyrifos (CPF) and its active metabolite, CPF-oxon (CPFO), in a human-derived liver cell line (HepG2) and kidney epithelial cell line (HK−2). The cytotoxicity to CPF and CPFO differed between cell lines where HK-2 had lower IC50 values which could be attributed to lower basal expression and inducibility of metabolizing enzymes, transporters, and nuclear receptors in HK-2 cells. In HepG2 cells, co-exposure of CPF with a specific inhibitor of either P-gp or BCRP enhanced the cytotoxicity of CPF while co-exposure of CPFO with VRP enhanced the cytotoxicity of CPFO, suggesting the role of these transporters in the elimination CPF and CPFO. Inhibition of efflux transporters did not affect the cytotoxicity of CPF and CPFO in HK-2 cells. Co-incubation of CPF with P-gp and BCRP inhibitors increased the intracellular concentration of CPF in HepG2 cells suggesting that both transporters play a role in limiting the cellular accumulation of CPF in HepG2 cells. Our results provide evidence that inhibition of efflux transporters can enhance CPF-induced toxicity through enhanced cellular accumulation and raises additional questions regarding how pesticide-transporter interactions may influence toxicity of mixtures containing pesticides and other environmental chemicals.

THE CYTOTOXIC EFFECT OF BENZOIC ACID ON TEN DIFFERENT CANCER CELL LINES

Cancer, having numerous types, is among the most dangerous and complex chronic diseases in the world affecting the wellbeing of humans, society and economy. The exploration and reassessment of effective chemicals, compounds, and natural products as potential agents for alleviating the adverse effects of cancer and its related symptoms continue on a global scale. This process involves an initial evaluation of the cytotoxic activities of potential drug candidates or treatment regimens on diverse cancer cell types in an ex vivo context. Benzoic acid (BA), an aromatic carboxylic acid that is widely available and used in the food industry, is one of the phenolic acids that may bear considerable anti-cancer potential. It is useful to find out the comparable effect of BA on various cancer types. Therefore, in this study, we tested the cytotoxicity of BA using MTT assay, on a number of ten different cancer cell lines and one normal cell type, namely prostate cancer (PC3), cervical cancer (HeLA), liver cancer (HUH7), colon cancer (CaCO2, HT29, SW48), bone cancer (MG63 and A673), pharyngeal cancer (2A3), lung cancer (CRM612) and kidney epithelial control cell line (Phoenix), respectively. IC50 (µg/ml) values after 48 and 72-hour exposure to BA were found to differ between 85.54±3.17 to 670.6±43.26, while the IC values for the control cell line Phoenix were 410.54±32.29 and 231.16±25.25, respectively. Taking into account of statistical evaluation of the IC50 values for BA on 11 cell types, we suggest that the molecular and omics approaches can be implemented in more details in order to find cellular and biochemical targets of BA as well as elucidating molecular mode of action, especially starting with the cancer cell lines of MG63, CRM612 and A673, in which the IC50 levels are relatively the lowest compared to those of the control cell line.

Building in vitro tools for livestock genomics: chromosomal variation within the PK15 cell line

BackgroundCultured porcine cell lines are powerful tools for functional genomics and in vitro phenotypic testing of candidate causal variants. However, to be utilised for genomic or variant interrogation assays, the genome sequence and structure of cultured cell lines must be realised. In this work, we called variants and used read coverage in combination with within-sample allele frequency to detect potential aneuploidy in two immortalised porcine kidney epithelial (PK15) cell lines and in a pig embryonic fibroblast line.ResultsWe compared two PK15 cultured cells samples: a new American Type Culture Collection (ATCC) sample and one that has been utilised and passaged within the laboratory for an extended period (> 10 years). Read coverage and within-sample allele frequencies showed that several chromosomes are fully or partially aneuploid in both PK15 lines, including potential trisomy of chromosome 4 and tetrasomy of chromosome 17. The older PK15 line showed evidence of additional structural variation and potentially clonal variation. By comparison, the pig embryonic fibroblast line was free from the gross aneuploidies seen in the PK15s.ConclusionsOur results show that the PK15 cell lines examined have aneuploidies and complex structural variants in their genomes. We propose that screening for aneuploidy should be considered for cell lines, and discuss implications for livestock genomics.

Stable in vitro fluorescence for enhanced live imaging of infection models for Batrachochytrium dendrobatidis.

Realistic and modifiable infection models are required to study the pathogenesis of amphibian chytridiomycosis. Understanding the mechanism by which Batrachochytrium dendrobatidis (Bd) can infect and kill diverse amphibians is key to mitigating this pathogen and preventing further loss of biodiversity. In vitro studies of Bd typically rely on a tryptone based growth media, whereas the recent development of a kidney cell-line infection model has provided a more realistic alternative, without the need for live animals. Here we use expression of a fluorescent reporter to enhance the in vitro cell-line based growth assay, and show that transformed Bd cells are able to invade and grow in an amphibian kidney epithelial cell line (A6) as well as in a new system using a lung fibroblast cell line (DWJ). Both Bd and host cells were modified to express reporter fluorescent proteins, enabling immediate and continuous observation of the infection process without the need for destructive sampling for fixation and staining. Plasmid DNA conferring hygromycin resistance and TdTomato (RFP) expression was delivered to Bd zoospores via electroporation, and continuous antibiotic selection after recovery produced stable fluorescent Bd transformants. Host cells (A6 and DWJ) were transfected before each assay using lipofection to deliver plasmid DNA conferring green fluorescent protein (GFP) and containing an empty shRNA expression cassette. Bd RFP expression allowed easy localisation of fungal cells and identification of endobiotic growth was assisted by host GFP expression, by allowing visualization of the space in the host cell occupied by the invading fungal body. In addition to enabling enhanced live imaging, these methods will facilitate future genetic modification and characterisation of specific genes and their effect on Bd virulence.

Comprehensive assessment of per and polyfluoroalkyl substances (PFAS) contamination in groundwater of Kamrup, Assam, India: occurrence, health risks, and metabolomic insights.

Per-and polyfluoroalkyl substances (PFAS) are synthetic chemicals that are known for their environmental persistence and adverse health effects. This study comprehensively assessed PFAS contamination in the Kamrup region of Assam, India, focusing on its presence in groundwater and associated health risks. The analysis detected 12 PFAS in groundwater samples from both the Kamrup Metro and Rural regions. In Kamrup Rural, Perfluorohexanoic acid (PFHxA), perfluorononanoic acid (PFNA), and perfluorooctanesulfonic acid (PFOS) were prevalent, whereas in Kamrup Metro, PFNA and PFOS were dominant, based on detection frequencies. These findings are noteworthy, as they demonstrate the widespread presence of PFAS in groundwater, a vital source of drinking water in the region. The assessment of PFAS health risks in India involved hazard quotient calculations for different age groups. Perfluorobutanesulfonic acid (PFBS) posed the highest risk, ranking children > boys > men > girls > women. Overall, ∑PFAS had low hazard (HQ: 0.27-0.41). Further, this study assessed PFBS and PFOS toxicity in human kidney epithelial cell lines (HEK293T) cells, revealing that PFBS was more cytotoxic than PFOS. The study examined the metabolomics of HEK293T cells after PFBS exposure, revealing significant alterations in lipid metabolism, particularly glycerophospholipids, potentially affecting cellular function and health. These findings underscore the importance of monitoring PFAS contamination in drinking water sources, especially in regions such as Kamrup, where groundwater is a primary source. Our metabolomics results show significant health effects at the cellular level, raising concerns about the impact of PFAS exposure on human health. This study highlights PFAS contamination in Kamrup, Assam's groundwater and its health risks, providing valuable insights for policymakers and public health management.

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.

Epigallocatechin-3-gallate attenuates arsenic-induced fibrogenic changes in human kidney epithelial cells through reversal of epigenetic aberrations and antioxidant activities.

Renal fibrosis is a pathogenic intermediate stage of chronic kidney disease (CKD). Nephrotoxicants including arsenic can cause kidney fibrosis through induction of oxidative stress and epigenetic aberrations. Epigallocatechin-3-gallate (EGCG), a green tea polyphenol, is known to have antioxidant and epigenetic modulation properties. Whether EGCG, through its antioxidant and epigenetic modulating activities, can attenuate fibrogenesis is not known. Therefore, the objective of this study was to determine whether EGCG can attenuate arsenic-induced acute injury and long-term exposure associated fibrogenicity in kidney epithelial cells. To address this question, two human kidney epithelial cell lines Caki-1 and HK-2 exposed to arsenic for both acute and long-term durations were treated with EGCG. The protective effect of EGCG on arsenic-induced cytotoxicity and fibrogenicity were evaluated by measuring the cell growth, reactive oxygen species (ROS) production, genes expression, and epigenetic changes in histone marks. Results revealed that EGCG has a protective effect in arsenic-induced acute cytotoxicity in these cells. EGCG scavenges the increased levels of ROS in arsenic exposed cells. Aberrant expression of fibrogenic genes in arsenic exposed cells were restored by EGCG. Abrogation of arsenic-induced fibrogenic changes was also associated with EGCG-mediated restoration of arsenic-induced aberrant expression of epigenetic regulatory proteins and histone marks. Novel findings of this study suggest that EGCG, through its antioxidant and epigenetic modulation capacities, has protective effects against arsenic-induced cytotoxicity and fibrogenic changes in kidney epithelial cells.

In-vitro Safety Assessment of Ultrasmall Gold Nanoparticles for Preclinical Drug Delivery Applications

Background: The development of safe and biocompatible nanoparticles has always been a major concern in nanomedicine applications. Various studies on the size-dependent toxicity of na-noparticles have been reported but are still controversial. The potential of small-sized nanoparticles can be utilized for imaging and diagnostics. However, insufficient toxicity data on these nanoparti-cles prevents researchers from utilizing their potential in diagnostics. More studies are needed on the toxicity of small-sized nanoparticles to present unanimous report for safe systemic use. The pre-sent study aimed to investigate the toxicity concerns of very small-sized AuNPs (2 ± 0.5 nm, 5 ± 1 nm, and 10 ± 2 nm) and provide a platform for their safe in vivo use. Methods: The cellular interactions of these three small-sized AuNPs with regard to cytotoxicity were investigated on hepatocellular carcinoma (HepG2) and epithelial kidney (HEK-293) cell lines. The cytotoxicity investigation of both cell lines was done through MTT assays, PI & DAPI, and cy-tology. Cellular stress was investigated by Catalase, TBARS, GSH, SOD & ROS parameters. The AuNPs incubated cells were also assessed for immunogenicity by ELISA, protein interaction by BSA, and cellular internalization by TEM (Edax). Results: All three-sized AuNPs were not toxic on cell viability, apoptosis, necrosis, or cytology as-sessment. No oxidative stress was noted in both cell types in the presence of 2 and 5-nm-sized AuNPs, whereas 10 nm-sized AuNPs showed little oxidative stress. AuNPs of size 2 and 5 nm were immunologically inert, but 10 nm-sized AuNPs elicited interleukin (IL-4 and IL-10) and interferon IFN gamma response. AuNPs of sized 2 nm showed 4 times the adsorption of albumin protein as compared to AuNPs of sized 5 nm. The TEM micrographs and peak of gold in the Edax graph con-firmed the presence of AuNPs in cells. Conclusion: Our results are suggestive of utilizing the potential of these three-sized AuNPs safely in preclinical drug delivery applications.

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.

Toxicity evaluation of synthetic glucocorticoids against breast cancer cell lines MDA-MB-231, MCF-7 and human embryonic kidney HEK293.

In this study, we conducted a comprehensive assessment of the cytotoxicity of three glucocorticoids, namely Hydrocortisone, Dexamethasone, and Methylprednisolone, using three different human cell lines: MDA-MB-231, MCF-7 (both adenocarcinoma cell lines), and HEK293 (kidney epithelial cell line). At lower concentrations exceeding 50µM, we did not observe any significant toxic effects of these glucocorticoids. However, when exposed to higher concentrations, Hydrocortisone exhibited dose-dependent cytotoxic effects on all three cell lines, with calculated IC50 values of 12 ± 0.6mM for HEK293, 2.11 ± 0.05mM for MDA-MB-231, and 2.73 ± 0.128mM for MCF-7 cells after 48h of exposure. Notably, Hydrocortisone, at its respective IC50 concentrations, demonstrated an inhibitory effect on the proliferation of the cancer cell lines, as evidenced by a substantial reduction in BrdU absorbance in a dose-dependent manner, coupled with a markedly reduced rate of colony formation in treated cells. Furthermore, Hydrocortisone exhibited remarkable anti-migratory properties in MDA-MB-231 and MCF-7 cells, and it induced cell cycle arrest in the SubG1 phase in MDA-MB-231 cells. In addition to these effects, Hydrocortisone triggered apoptosis in both cancer cell types, leading to observable morphological changes. This apoptotic response was characterized by a significant increase in the activity of caspase-3, which was time-dependent. Additionally, Hydrocortisone downregulated the expression of anti-apoptotic Bcl-2 proteins. In summary, our findings underscore the safety of clinical doses in terms of cell toxicity meanwhile increased concentration were showing an anti-proliferative potential of Hydrocortisone, particularly against adenocarcinoma breast cancer cell lines.

RhMYDGF Alleviates I/R-induced Kidney Injury by Inhibiting Inflammation and Apoptosis via the Akt Pathway.

Renal ischemia/reperfusion (I/R) injury is one of the crucial factors affecting the outcome of renal transplantation. In recent years, myeloid-derived growth factor (MYDGF) has received a lot of attention for its extensive beneficial effects on cardiac repair and protection of cardiomyocytes from cell death. Therefore, we hypothesized that the recombinant human MYDGF (rhMYDGF) protein might play an essential role in safeguarding renal I/R injury. In vivo experiments were conducted using a mouse unilateral I/R model. Mice were pretreated with rhMYDGF by intraperitoneal injection to study the potential mechanism of renal protection. In vitro, we established hypoxia/reoxygenation and H 2 O 2 treatment models to pretreat cells with rhMYDGF. The expression levels of oxidative stress, inflammation, and apoptosis-related factors in tissues and cells were detected. Finally, we explored the role of the protein kinase B (Akt) pathway in the renal protective mechanism of rhMYDGF. In this study, we found that intraperitoneal injection of 1.25 μg rhMYDGF could significantly improve renal function of I/R mice, and reduce oxidative stress, inflammation, and apoptosis. For the human proximal tubular epithelial cell line and human kidney cell line, pretreatment with 0.3 μg/mL rhMYDGF for 24 h significantly downregulated oxidative stress, inflammation, and apoptosis via the phosphorylation of Akt, which could be ameliorated by LY294002. rhMYDGF protects kidney from I/R injury by attenuating oxidative stress, inflammation, and apoptosis through the activation of the Akt pathway.

SETD2 loss in renal epithelial cells drives epithelial-to-mesenchymal transition in a TGF-β-independent manner.

Histone-lysine N-methyltransferase SETD2 (SETD2), the sole histone methyltransferase that catalyzes trimethylation of lysine 36 on histone H3 (H3K36me3), is often mutated in clear cell renal cell carcinoma (ccRCC). SETD2 mutation and/or loss of H3K36me3 is linked to metastasis and poor outcome in ccRCC patients. Epithelial-to-mesenchymal transition (EMT) is a major pathway that drives invasion and metastasis in various cancer types. Here, using novel kidney epithelial cell lines isogenic for SETD2, we discovered that SETD2 inactivation drives EMT and promotes migration, invasion, and stemness in a transforming growth factor-beta-independent manner. This newly identified EMT program is triggered in part through secreted factors, including cytokines and growth factors, and through transcriptional reprogramming. RNA-seq and assay for transposase-accessible chromatin sequencing uncovered key transcription factors upregulated upon SETD2 loss, including SOX2, POU2F2 (OCT2), and PRRX1, that could individually drive EMT and stemness phenotypes in SETD2 wild-type (WT) cells. Public expression data from SETD2 WT/mutant ccRCC support the EMT transcriptional signatures derived from cell line models. In summary, our studies reveal that SETD2 is a key regulator of EMT phenotypes through cell-intrinsic and cell-extrinsic mechanisms that help explain the association between SETD2 loss and ccRCC metastasis.

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.

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.

Antiviral effects and tissue exposure of tetrandrine against SARS-CoV-2 infection and COVID-19.

Tetrandrine (TET) has been used to treat silicosis in China for decades. The aim of this study was to facilitate rational repurposing of TET against SARS-CoV-2 infection. In this study, we confirmed that TET exhibited antiviral potency against SARS-CoV-2 in the African green monkey kidney (Vero E6), human hepatocarcinoma (Huh7), and human lung adenocarcinoma epithelial (Calu-3) cell lines. TET functioned during the early-entry stage of SARS-CoV-2 and impeded intracellular trafficking of the virus from early endosomes to endolysosomes. An in vivo study that used adenovirus (AdV) 5-human angiotensin-converting enzyme 2 (hACE2)-transduced mice showed that although TET did not reduce pulmonary viral load, it significantly alleviated pathological damage in SARS-CoV-2-infected murine lungs. The systemic preclinical pharmacokinetics were investigated based on in vivo and in vitro models, and the route-dependent biodistribution of TET was explored. TET had a large volume of distribution, which contributed to its high tissue accumulation. Inhaled administration helped TET target the lung and reduced its exposure to other tissues, which mitigated its off-target toxicity. Based on the available human pharmacokinetic data, it appeared feasible to achieve an unbound TET 90% maximal effective concentration (EC90) in human lungs. This study provides insights into the route-dependent pulmonary biodistribution of TET associated with its efficacy.

Identification of an unrecognized circRNA associated with development of renal fibrosis.

Backgroud: Renal fibrosis is the common characteristic of chronic kidney disease. Circular RNA plays an essential role in the occurrence and development of Renal fibrosis, but its regulative mechanism remains elusive. Methods: The animal and cell model of Renal fibrosis was established, and RNA-sequencing and real-time polymerase chain reaction (qRT-PCR) experiments were implemented. Subsequently, experiments for detecting apoptosis and proliferation of cell, were carried out, and the isobaric tags for relative and absolute quantification proteomics analyses were performed accordingly. Results: It was found that a newly discovered Circular RNA (circRNA_0002158), is highly expressed in kidneys or cells with fibrosis, implying that this Circular RNA might be associated with the occurrence and development of Renal fibrosis. Subsequently, the overexpression and knockdown of circRNA_0002158 were conducted in the human kidney epithelial cell line (HK-2) cells, and the results indicated that the circRNA_0002158 could inhibit apoptosis, and promote proliferation of cells. The kidney injury-related factors, including Fibronectin and plasminogen activator inhibitor-1 (PAI-1), were decreased in HK-2 cells with overexpression of circRNA_0002158, while the results were reversed in cells with knockdown of circRNA_0002158. Finally, to explore the regulative mechanism of circRNA_0002158, the iTRAQ proteomics analyses were implemented for the cell samples with OE of circRNA_0002158 and its control, it showed that multiple genes and functional pathways were associated with the occurrence and development of Renal fibrosis. Conclusion: CircRNA_0002158 is associated with regulating Renal fibrosis, and may contribute to ameliorating the progression of Renal fibrosis in the future.

A biopolymer functionalized two-dimensional WS2@TiO2 nanocomposite for in vitro biomedical and photocatalytic applications: facile synthesis and characterization.

Organic dyes and microorganisms in industrial wastewater have harmed both the environment and human health. In this present study, the in vitro biological and photocatalytic properties of a synthesized biogenic chitosan functionalized WS2@TiO2 hybrid nanocomposite (NC) are investigated. The chitosan-functionalized WS2@TiO2 hybrid nanocomposite (NC) was synthesized hydrothermally. Its microstructure and compositional properties were studied. The antibacterial activity against Staphylococcus aureus, and Bacillus subtilis (Gram-positive) and Klebsiella pneumoniae, and Escherichia coli (Gram-negative) was evaluated. The NC exhibits the highest antibacterial activity against K. pneumoniae at bacterial inhibition zones of 27 mm. It also showed remarkable anticancer effects in MCF-7 cells (cell inhibition of 74% at 100 μg mL-1). The biocompatibility of the composite was tested against the Vero (kidney epithelial) cell line. The results suggest that the NC had no obvious cytotoxicity. Also, the NC showed good photocatalytic performance (degradation rate of 89.43% at 150 min; K = 0.0175 min-1). The results suggest that chitosan functionalized WS2@TiO2 NCs are a potential candidate for biological and environmental applications.

Synthesis, conformation and cytotoxic activity of short hybrid peptides containing conformationally constrained 1-(aminomethyl)cyclohexanecarboxylic acid and gabapentin

The present work describes the synthesis,conformation and cytotoxic activities of short β/γ hybrid peptides, Boc-β2,2-Ac6c-Gpn-NHMe, BG1; Boc-(β2,2-Ac6c-Gpn)2-OMe, BG2; Boc-(β2,2-Ac6c-Gpn)3-OMe, BG3; H-β2,2-Ac6c-Gpn-NHMe, BG4; H-(β2,2-Ac6c-Gpn)2-OMe, BG5; H-(β2,2-Ac6c-Gpn)3-OMe, BG6, Boc-β2,2-Ac6c-Gpn-OMe, BG7 and H-β2,2-Ac6c-Gpn-OMe, BG8. Mixed C6/C7 conformations were observed for β/γ hybrid peptides. Further, BG1-BG8 were screened against MCF-7 (Breast cancer), A549 (Lung Cancer), PC-3 (Prostate cancer), HCT-116 (Colon cancer), and MDA-MB-231 (Breast cancer) cell lines. Among all, BG6 exhibited potent cytotoxicity against all cancer cell lines with IC50 ranging from 1.6 μM to 6.3 μM with relatively low cytotoxicity against normal epithelial breast cell line fR-2 and human embryonic kidney cell line HEK-293. Minimal hemolytic activity was observed for BG6 against human erythrocytes. Peptide BG6 displayed anti-migratory and anti-invasive potentials showing strong interactions with intrinsic apoptotic markers Bcl-2, Bax, and cleaved-PARP, as well as the induction of the mitochondria maladjustment mediated apoptosis.

Isolation, Propagation and Genotyping of Human Rotaviruses Circulating among Children with Gastroenteritis in Two Egyptian University Hospitals.

Simple SummaryGroup A rotaviruses are the most common cause of acute gastroenteritis affecting Egyptian children under the age of five, with symptoms ranging from asymptomatic infection to severe dehydration or death. In the present work, diarrheal samples from Egyptian children admitted to gastrointestinal pediatric wards of two main governmental hospitals were collected and molecularly analyzed for Group A rotavirus. Our findings revealed that rotaviruses accounted for more than one-sixth of all cases under study, peaking in the winter. G1P[8] was the most prevalent rotavirus genotype in this study. The two cell lines used in our work coherently isolated and propagated rotavirus strains. Continuous rotavirus detection and genome sequencing of the successfully isolated strains will be recommended in the future in order to support the control of such viruses, and tackle the problem in Egypt.The most prevalent cause of infectious neonatal diarrhea is Group A rotavirus (RVA). Unfortunately, there is a dearth of data on the incidence of rotavirus-associated infections among Egyptian children. The present study aimed to isolate, propagate, and genotype human rotaviruses circulating among Egyptian children with acute gastroenteritis admitted to two main university pediatric hospitals, Abo El-Reesh and El-Demerdash, over two consecutive winters, 2018–2020. Diarrheal samples (n = 230) were screened for Group A rotavirus RNA using RT-PCR assay. In positive samples (n = 34), multiplex semi-nested PCR was utilized to determine G and P genotypes. Thirty-four (14.8%) of the collected samples tested positive. The genotype distribution revealed that G1P[8] was the predominant rotavirus genotype throughout the current study. All rotavirus-positive fecal samples were passaged twice on human colorectal adenocarcinoma cell line (Caco-2) and rhesus monkey kidney epithelial cell line (MA104). Both cell lines could successfully isolate 14.7% (n = 5 out of 34) of the identified strains; however, Caco-2 cell line was shown to be more efficient than MA104 in promoting the propagation of human rotaviruses identified in Egyptian children’s feces.

Inhibition of kidney potassium channels by fluoxetine: In vivo and in vitro studies.

In vitro studies have demonstrated that fluoxetine, a commonly used antidepressant drug, can modulate the activity of K+ channels. In the present study, we investigated the in vivo effect of acute and sub-chronic treatment of rats with fluoxetine on K+ renal transport. Furthermore, OK cells, a kidney epithelial cell line, were used in order to evaluate the in vitro effect of fluoxetine on K+ currents. In the sub-chronic study, fluoxetine was administrated daily (10mg/kg, p.o.) for 15 days to male adult Wistar rats. In the acute study, rats were given increasing doses of fluoxetine (1, 3, 10, 30 and 50 mg/kg, p.o.) for 24 h. Results from the sub-chronic study show that urinary K+ content (in mmol/L) was markedly reduced in the fluoxetine-treated animals (fluoxetine: 83 ± 9; control: 131 ± 10; P < 0.001). K+ fractional renal excretion (in %) was also significantly lower in the fluoxetine group (fluoxetine: 6 ± 1; control: 13 ± 2; P < 0.001). No significant changes was observed in creatinine clearance and on renal tubular Na+ ,K+ -ATPase activity. Results obtained from the acute study demonstrate that, after a 24-h administration, fluoxetine produced a dose-dependent decrease in urinary K+ , with an ED50 (in mg/kg) of 4.2 (2.8; 5.5) and a maximal effect of 62% reduction. In vitro, fluoxetine produced a concentration-dependent inhibition of K+ currents in OK cells, with an EC50 of 107 (84.8; 129.5) μM. In conclusion, fluoxetine produces a marked reduction on urinary K+ excretion; this effect constitutes an in vivo evidence for the inhibitory action of fluoxetine on kidney epithelial K+ channels.