MDA Cell Research Articles

Effects of dietary Lactobacillus postbiotics and bacitracin on the modulation of mucosa-associated microbiota and pattern recognition receptors affecting immunocompetence of jejunal mucosa in pigs challenged with enterotoxigenic F18+ Escherichia coli

BackgroundEnterotoxigenic Escherichia coli (E. coli) is a threat to humans and animals that causes intestinal disorders. Antimicrobial resistance has urged alternatives, including Lactobacillus postbiotics, to mitigate the effects of enterotoxigenic E. coli.MethodsForty-eight newly weaned pigs were allotted to NC: no challenge/no supplement; PC: F18+ E. coli challenge/no supplement; ATB: F18+ E. coli challenge/bacitracin; and LPB: F18+ E. coli challenge/postbiotics and fed diets for 28 d. On d 7, pigs were orally inoculated with F18+ E. coli. At d 28, the mucosa-associated microbiota, immune and oxidative stress status, intestinal morphology, the gene expression of pattern recognition receptors (PRR), and intestinal barrier function were measured. Data were analyzed using the MIXED procedure in SAS 9.4.ResultsPC increased (P < 0.05) Helicobacter mastomyrinus whereas reduced (P < 0.05) Prevotella copri and P. stercorea compared to NC. The LPB increased (P < 0.05) P. stercorea and Dialister succinatiphilus compared with PC. The ATB increased (P < 0.05) Propionibacterium acnes, Corynebacterium glutamicum, and Sphingomonas pseudosanguinis compared to PC. The PC tended to reduce (P = 0.054) PGLYRP4 and increased (P < 0.05) TLR4, CD14, MDA, and crypt cell proliferation compared with NC. The ATB reduced (P < 0.05) NOD1 compared with PC. The LPB increased (P < 0.05) PGLYRP4, and interferon-γ and reduced (P < 0.05) NOD1 compared with PC. The ATB and LPB reduced (P < 0.05) TNF-α and MDA compared with PC.ConclusionsThe F18+ E. coli challenge compromised intestinal health. Bacitracin increased beneficial bacteria showing a trend towards increasing the intestinal barrier function, possibly by reducing the expression of PRR genes. Lactobacillus postbiotics enhanced the immunocompetence of nursery pigs by increasing the expression of interferon-γ and PGLYRP4, and by reducing TLR4, NOD1, and CD14.

Novel Tetracyclic Azaphenothiazines with the Quinoline Ring as New Anticancer and Antibacterial Derivatives of Chlorpromazine.

Phenothiazine derivatives are widely studied in various fields such as biology, chemistry, and medicine research because of their pharmaceutical effects. The first compound used successfully in the treatment of psychosis was a phenthiazine derivative, chlorpromazine. Apart from its activity in neurons, chlorpromazine has also been reported to display anticancer and antibacterial properties. In this study, we present the synthesis and research on the activity of A549, MDA, MiaPaCa, PC3, and HCT116 cancer cell lines and of S. aureus, S. epidermidis, E. coli, and P. aeruginosa bacterial strains against a series of new tetracyclic chlorpromazine analogues containing a quinoline scaffold in their structure instead of the benzene ring and various substituents at the thiazine nitrogen. The structure of these novel molecules has been determined by 1H NMR, 13C NMR, and HRMS spectral techniques. The seven most active of the twenty-four new chlorpromazine analogues tested were selected to study the mechanism of cytotoxic action. Their ability to induce apoptosis or necrosis in cancer cells was assessed by flow cytometry analysis. The results obtained confirmed the proapoptotic activity of selected compounds, especially in terms of inducing late apoptosis or necrosis in cancer cell lines A549, MiaPaCa-2, and HCT-116. Furthermore, studies on the induction of cell cycle arrest suggest that the new chlorpromazine analogues exert antiproliferative effects by inducing cell cycle arrest in the S phase and, consequently, apoptosis.

Physicochemical Properties and Biological Activities of Quinoa Polysaccharides.

Quinoa, known as the "golden grain" for its high nutritional value, has polysaccharides as one of its sources of important nutrients. However, the biological functions of quinoa polysaccharides remain understudied. In this study, two crude polysaccharide extracts of quinoa (Q-40 and Q-60) were obtained through sequential precipitation with 40% and 60% ethanol, with purities of 58.29% (HPLC) and 62.15% (HPLC) and a protein content of 8.27% and 9.60%, respectively. Monosaccharide analysis revealed that Q-40 contained glucose (Glc), galacturonic acid (GalA), and arabinose (Ara) in a molar ratio of 0.967:0.027:0.006. Q-60 was composed of xylose (xyl), arabinose (Ara), galactose, and galacturonic acid (GalA) with a molar ratio of 0.889:0.036:0.034:0.020. The average molecular weight of Q-40 ranged from 47,484 to 626,488 Da, while Q-60 showed a range of 10,025 to 47,990 Da. Rheological experiments showed that Q-40 exhibited higher viscosity, while Q-60 demonstrated more elastic properties. Remarkably, Q-60 showed potent antioxidant abilities, with scavenging rates of 98.49% for DPPH and 57.5% for ABTS. Antibacterial experiments using the microdilution method revealed that Q-40 inhibited the growth of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), while Q-60 specifically inhibited MRSA. At lower concentrations, both polysaccharides inhibited MDA (MD Anderson Cancer Center) cell proliferation, but at higher concentrations, they promoted proliferation. Similar proliferation-promoting effects were observed in HepG2 cells. The research provides important information in the application of quinoa in the food and functional food industries.

LncRNA MEG3 suppresses erastin-induced ferroptosis of chondrocytes via regulating miR-885-5p/SLC7A11 axis.

Ferroptosis is involved in osteoarthritis development; however, the roles of long noncoding RNAs (lncRNAs), including lncRNA MEG3, in the regulation of ferroptosis in osteoarthritis are still unclear. In this study, qRT‒PCR and Western blotting assays were used to detect the expression of lncRNA MEG3, miR-885-5p, SLC7A11 and GPX4; MDA and CCK-8 assays were applied to analyse cellular MDA levels and cell viability, respectively. Erastin elevated cellular MDA levels and decreased the viability of chondrocytes and the erastin-induced decline in cell viability was reversed by a ferroptosis inhibitor (ferrostatin-1). Erastin downregulated lncRNA MEG3, SLC7A11 and GPX4 and upregulated miR-885-5p. Silencing of lncRNA MEG3 increased miR-885-5p and downregulated SLC7A11 and GPX4 and further sensitized chondrocytes to erastin-induced ferroptosis. In contrast, overexpression of lncRNA MEG3 had opposite effects. Dual luciferase assays confirmed binding between lncRNA MEG3 and miR-885-5p and between miR-885-5p and the 3'UTR of SLC7A11. In the synovial fluids from patients with osteoarthritis compared with synovial fluids from normal controls, the RNA levels of lncRNA MEG3 and SLC7A11 were decreased and the miR-885-5p expression level was increased. Our findings indicated that lncRNA MEG3 overexpression alleviated ferroptosis in chondrocytes by affecting the miR-885-5p/SLC7A11 signalling pathway.

Preclinical and dose-ranging assessment of hESC-derived dopaminergic progenitors for a clinical trial on Parkinson's disease

Human embryonic stem cell (hESC)-derived midbrain dopaminergic (mDA) cell transplantation is a promising therapeutic strategy for Parkinson's disease (PD). Here, we present the derivation of high-purity mDA progenitors from clinical-grade hESCs on a large scale under rigorous good manufacturing practice (GMP) conditions. We also assessed the toxicity, biodistribution, and tumorigenicity of these cells in immunodeficient rats in good laboratory practice (GLP)-compliant facilities. Various doses of mDA progenitors were transplanted into hemi-parkinsonian rats, and a significant dose-dependent behavioral improvement was observed with a minimal effective dose range of 5,000-10,000 mDA progenitor cells. These results provided insights into determining a low cell dosage (3.15 million cells) for human clinical trials. Based on these results, approval for a phase 1/2a clinical trial for PD cell therapy was obtained from the Ministry of Food and Drug Safety in Korea, and a clinical trial for treating patients with PD has commenced.

The Newly Proposed Mechanism of Cardiomyocyte Protection of Carvedilol- Anti-Apoptosis Pattern of Carvedilol in Anoxia by Inducing Autophagy Partly through the AMPK/mTOR Pathway

Purpose: To investigate the underlying mechanism of cardiomyocyte protection of carvedilol based on autophagy and apoptosis. Methods: Neonatal rat ventricular myocytes (NRVMs) were exposed to various concentrations of carvedilol before anoxia, and pretreated with 3-MA or compound C for inhibiting autophagy or p-AMPK expression. CCK-8 colorimeter and flow cytometry were used to determine the cell viability and apoptotic rates. The variation of mRNA and protein was measured by RT-PCR and Western blot. The presence of autophagosomes was observed by electron microscopy. Results: First, we found that carvedilol increased autophagic marker levels in a concentration-dependent manner and the number of autophagosomes in NRVMs. Moreover, carvedilol substantially enhanced the viability and noticeably reduced the CK, MDA and LDH levels and cell apoptosis rate compared with the anoxia group. In addition, carvedilol decreased the levels of caspase-3 and Bim in mRNA and protein, but such effect was blocked by the special autophagy inhibitor-3-MA, and the number of autophagosomes was significantly decreased when treated with 3-MA, indicating that carvedilol exhibited anti-apoptotic and anti-injury effects by inducing autophagy in anoxia NRVMs, but these effects can be abolished by adding 3-MA to suppress autophagy. Finally, the carvedilol treatment-induced autophagy by enhancing the activation of p-AMPK and inhibiting p-mTOR. Electron microscopy presented that the number of autophagosomes was significantly decreased when treating with compound C, indicating that carvedilol induced autophagy in anoxia NRVMs partly by the AMPK-mTOR signaling pathway. Conclusions: Carvedilol has cardioprotection by inducing autophagy against apoptosis partly through the AMPK/mTOR pathway during anoxia in NRVMs.

Antibody-Free Fluorine-Assisted Metabolic Sequencing of RNA N4-Acetylcytidine.

N4-Acetylcytidine (ac4C) has been found to affect a variety of cellular and biological processes. For a mechanistic understanding of the roles of ac4C in biology and disease, we present an antibody-free, fluorine-assisted metabolic sequencing method to detect RNA ac4C, called "FAM-seq". We successfully applied FAM-seq to profile ac4C landscapes in human 293T, HeLa, and MDA cell lines in parallel with the reported acRIP-seq method. By comparison with the classic ac4C antibody sequencing method, we found that FAM-seq is a convenient and reliable method for transcriptome-wide mapping of ac4C. Because this method holds promise for detecting nascent RNA ac4C modifications, we further investigated the role of ac4C in regulating chemotherapy drug resistance in chronic myeloid leukemia. The results indicated that drug development or combination therapy could be enhanced by appreciating the key role of ac4C modification in cancer therapy.

Synthesis, biological evaluation and in silico studies of novel thiadiazole-hydrazone derivatives for carbonic anhydrase inhibitory and anticancer activities

ABSTRACT Thiadiazole and hydrazone derivatives (5a–5i) were synthesized and their chemical structures were verified and described by 1H NMR, 13C NMR, and HRMS spectra. Three cancer cell lines (MCF-7, MDA, and HT-29) and one healthy cell line (L929) were used to test the cytotoxicity activity of synthesized compounds as well as their inhibitory activity against carbonic anhydrase I, II and IX isoenzymes. Compound 5d (29.74 µM) had a high inhibitory effect on hCA I and compound 5b (23.18 µM) had a high inhibitory effect on hCA II. Furthermore, compound 5i was found to be the most potent against CA IX. Compounds 5a-5i, 5b and 5i showed the highest anticancer effect against MCF-7 cell line with an IC50 value of 9.19 and 23.50 µM, and compound 5d showed the highest anticancer effect against MDA cell line with an IC50 value of 10.43 µM. The presence of fluoro substituent in the o-position of the phenyl ring increases the effect on hCA II, while the methoxy group in the o-position of the phenyl ring increases the activity on hCA I as well as increase the anticancer activity. Cell death induction was evaluated by Annexin V assay and it was determined that these compounds cause cell death by apoptosis. Molecular docking was performed for compounds 5b and 5d to understand their biological interactions. The physical and ADME properties of compounds 5b and 5d were evaluated using SwissADME.

Phytomediated synthesis of bimetallic Ag/Au nanoparticles using orange peel extract and assessment of their antibacterial and anitcancer potential

The current research interest in nanoscience centers around the synthesis of nanoparticle with noble metals due to their distinctive characteristics that has expanded their prospective utilization in various fields of science and technology. The study reports the green fabrication of bimetallic composite nanoparticles (CNPs) with silver (Ag) and gold(Au) via an eco-friendly mode with a biomaterial, orange peel (OP) extract as a reductant. The reaction of aqueous solutions of silver nitrate (AgNO3) and chloroauric acid (HAuCl4) with the OP extract resulted in rapid production of stable CNPs having spectacular spherical shapes and other morphologies. The characterization of the synthesized CNPs included the following analyses; UV–vis spectrophotometer, (DLS) dynamic light scattering technique for the particles average size was found to be 138 nm with a polydispersity index (PDI) of 0.249. To identify the functional groups in biomolecules that emphasizes the reducing and capping agent for NPs, the Fourier transform infrared spectroscopy (FTIR) has been used. The characterization of as prepared NPs has been complemented with (TEM) transmission Electron Microscopy, and (SEM) scanning Electron Microscopy along with the (EDS) energy dispersive spectrometry for the elemental screening of the sample. Furthermore, antibacterial effect and the cytotoxicity activity against the human MDA cell line of the bimetallic CNPs were investigated. The key findings showed that the CNPs suspension possesses potent cytotoxic activity (and was highly effective against bacterial strains. Thus, the finding support the prospective therapeutic potential of the nanoparticles in nano medicine.

Transcriptome profiling uncovers the lncRNA-mediated regulatory networks associated with tolerance to cadmium stress in barley

Cadmium (Cd) is one of the most harmful and widespread toxic heavy metal contaminants. Long non-coding RNAs (lncRNAs) play vital roles in response to abiotic stress by participating in the gene regulatory network. Here, we performed a transcriptome analysis of lncRNAs in roots of the two contrasting barley genotypes, namely Zhenong8 (ZN, Cd-tolerant) and W6nk2 (Cd-sensitive). In total, 9937 novel lncRNAs were identified, and 5758 lncRNA-target pairs were predicted to be cis-acting and 4159 to be trans-acting. Further comparison of genotypic differences revealed 195 novel lncRNAs as differentially expressed between the two genotypes in response to Cd stress. The miRanda prediction and expression relationship analyses among miRNA, lncRNA, and mRNA revealed that 8 lncRNAs may act as target mimics of miRNAs in barley. Among them, HvGAMYB was predicted to be the target gene of miR319a_1 and miR319b-3p, with LTCONS_00177709 and LTCONS_00185175 acting as endogenous competitors. Barley stripe mosaic virus-induced gene silencing (BSMV-VIGS) of HvGAMYB in ZN8 led to hypersensitivity to Cd stress with significantly increased Cd concentration in both roots and shoots as well as Cd accumulation in shoots. Moreover, silencing HvGAMYB suppressed photosynthesis and antioxidant-related genes but stimulated the production of reactive oxygen species (ROS) after Cd exposure, leading to exacerbated Cd-induced MDA accumulation and cell death. Our findings provide new knowledge and insights into the regulatory functions of lncRNAs in plant Cd tolerance.

Production of Childinan SF-2 as Bioactive Compound from Daldinia childiae: A Survey on Antioxidant, Antibacterial and Antitumor Properties.

Fungal extracts have received increased attention due to their great medicinal applications including antitumor, immune-modulating, antioxidant, radical scavenging, antiviral, antibacterial, antifungal and detoxificating properties. This study is the first report on a novel bioactive compound, namely Childinan SF-2 which was isolated from soil ascomycete fungus. The significant antibacterial, antioxidant and cytotoxic properties of the extract may lead to development of novel, safe and useful substances. The isolate was identified on the basis of molecular approach. Spore suspension was inoculated in the culture medium and the bioactive compound was isolated from the viscous fermented broth via ethanol precipitation of the extracellular compound. The basic chemical composition of the extract including protein, carbohydrate, sulfate radical and uronic acid content were measured. FTIR (Fourier-transform infrared spectroscopy) and GC-MS (Gas chromatography-mass spectrometry) analysis were used for further structural characterization. The extract was utilized for treatment of AGS and MDA cell lines to assess the cell cycle and apoptosis. The antioxidant activity was examined using DPPH, hydroxyl radicals scavenging, β-carotene bleaching inhibition and ferric reducing power assay methods. The extract was tested for evaluation of antibacterial activity using different Gram-positive and Gram-negative bacterial strains. The fungal isolate was identified as the new strain Daldinia childiae SF-2. Initial biochemical characterization of the extract showed that the fungal biopolymer was composed of total sugars, protein, uronic acids and sulfated groups with values of 91.6%, 2.15%, 2.25% and 1.05% (w/w), respectively. FTIR and GC-MS analysis revealed that Childinan SF-2 might be mainly constructed from D-glucose, D-mannitol and D-galactofuranose. The in vitro experiments revealed that Childinan SF-2 enhanced the percentage of necrosis and apoptosis of cancer cells and blocked the cell cycle progression as shown by flowcytometry. Childinan SF-2 represented a considerable antioxidant and antibacterial activity. These results indicated that Childinan SF-2 can serve as a potential source in medicinal applications. Keywords Exopolysaccharides; Fungal Biopolymers; Mycelial Secretions; Natural Pharmaceuticals; Xylariaceae.

Therapeutic efficacy of Nano-formulation of lactoperoxidase and lactoferrin via promoting immunomodulatory and apoptotic effects

This study identifies promising potential of a novel and safer nanocombination of bovine milk lactoperoxidase (LPO) and lactoferrin (LF) to target breast cancer in vitro and in adult female albino rat model. Favorable selective anticancer effects of the prepared nanocombination were observed, in a dose-dependent manner, against both MCF-7 and MDA cell lines, sparing normal HFB-4 cells. The administration of LPO + LFNPs markedly improved the induced-breast cancer disorders, prolonged survival and reduced the values of serum TNF-α, IL1β, CD4+, ALAT, ASAT, urea, creatinine, cholesterol and triglycerides with remarkable elevation in mammary SOD and GPx activity and GSH level. Moreover, the histopathological findings showed that LPO + LFNPs succeeded in prevention of mammary gland tumorigenesis. Superior efficacy of LPO + LFNPs was observed against pro-inflammatory cytokines through their anti-inflammatory and immunomodulatory properties. The treatment of LPO + LFNPs more significantly modulated the apoptosis and enhanced the expression of cell cycle regulator genes, which demonstrates a successful tumor therapy in vitro and in vivo. Therefore, this study provided evidence that the chemo-preventive feature of LPO + LFNPs may offer a novel alternative therapy for the treatment of breast cancer through enhances apoptosis pathway, improvement of immune response, reduction of inflammation and restoration of the impaired oxidative stress.

A novobiocin derivative, XN4, triggers ferroptosis in gastric cancer cells via the activation of NOX4

Context A novobiocin derivative, XN4, has been shown to promote cell apoptosis in chronic myeloid leukaemia. Objective This study explores the mechanism by which XN4 promotes ferroptosis of gastric cancer (GC) cells. Materials and methods Human GC SGC-7901 and BGC-823 cells were treated with different XN4 concentrations (0, 0.1, 0.5, 1.0, 5.0, and 10.0 μmol/L) to evaluate effects of XN4. Additionally, cells were pre-treated for 24 h with si-NOX4, for 1 h with the iron chelator deferoxamine mesylate (DFO) or for 1 h with the lipid peroxidation inhibitor liproxstatin-1 before being treated with XN4 to analyse the mechanism of XN4. Results XN4 increased cell death (IC50 values of XN4 on SGC-7901 and BGC-823 cells: 1.592 ± 0.14 μmol/L and 2.022 ± 0.19 μmol/L) and Fe2+ levels in SGC-7901 and BGC-823 cells. These effects of 2.0 μmol/L XN4 were abolished by 100 μmol/L DFO treatment. XN4 enhanced transferrin and transferrin receptor expression to induce Fe2+ accumulation. XN4 decreased mitochondrial membrane potentials in GC cells, similar to erastin. Additionally, XN4 increased MDA, hydrogen peroxide, and ROS levels, but diminished total glutathione levels. Liproxstatin-1 (200 nmol/L) nullified the effects of XN4 (2.0 μmol/L) on MDA levels and cell death. Moreover, GPX4 levels decreased, but NOX4 and ferroptosis-related protein PTGS2 levels increased in GC cells following XN4 treatment, which was nullified by NOX4 knockdown. Discussion and conclusions The pro-ferroptotic role of XN4 in GC might enable it to become a promising drug for GC treatment in the future despite the need for extensive research.

Abstract 2875: Development of a new tumor-bearing humanized mouse model to evaluate the efficacy and toxicity of bispecific T cell engagers

Abstract Background: T cell-based immunotherapies, especially the bispecific T cell engager (BiTE), which recruit T cells to tumor cells, have been developed rapidly for treatment of malignancies. However, the wide application of BiTE therapies is limited by accompanying severe adverse effects such as cytokine release syndrome (CRS). Since human cancer cells are not incorporated in the CRS evaluation, currently used in vivo animal models and in vitro human immune cell activation assays do not reliably evaluate the induction of CRS by immune drugs that is seen in BiTE treated cancer patients. In addition, the differences between human and mouse immune systems hamper the development of new immune-oncology therapeutic drugs. Thus, establishing a predictive in vivo animal model that can screen both efficacy of cancer therapy and risk of developing CRS is critical for improving the safety of immune drug development for treatment of cancer patients. Method: Humanized mice were developed in NSG-MHC class I/II double knock out (DKO) mouse recipients following reconstitution with human peripheral blood mononuclear cells. The MDA-MB-231/Luciferase-2A-GFP stable breast cancer cell line (MDA), which expressed epidermal growth factor receptor (EGFR), were implanted in mice 5 days after PBMC engraftment. Following EGFRxCD3 BiTE treatment whole-body tumor burden was quantified by an in vivo imaging system. To evaluate the CRS levels in response to BiTE therapy, cytokine levels were analyzed by BD cytometric bead array. Clinical observations, CRS scores, body weight changes and human immune cell activation were also monitored for CRS assessment. Results: PBMC humanized models were successfully established with breast cancer cell engraftment. After treatment with EGFRxCD3 BiTE, the mice exhibited elevated serum levels of human IFNgamma, TNF-a, IL-6, IL-10, IL-2 and IL-4 compared to a vehicle control group. This cytokine production was dose-dependent and was associated with an increase of toxic mouse clinical evaluation score. In contrast, there was no significant inflammatory cytokine production in PBMC humanized mice without MDA cell implantation, suggesting T cell activation by BiTE followed binding to the cancer cells. BiTE treatment for 2-4 days significantly suppressed tumor growth in lung as measured by luciferin imaging. With serial doses of BiTE treatment, we observed a dose-dependent inhibition of tumor growth. Conclusion: We developed a novel in vivo PBMC humanized mouse model bearing human tumor cells that can be used to determine the optimal dose of BiTE for inhibition of tumor growth with low toxicity risk. This model provides a growth environment similar to that of patients and has important application values in simultaneously evaluating the efficacy and safety of any T cell-based immunotherapies including BiTE to define optimal approaches for clinical cancer treatment. Citation Format: Guoxiang Yang, Li-Chin Yao, Michael A. Brehm, Dale L. Greiner, Leonard D. Shultz, Danying Cai, MIngshan Cheng, James G. Keck. Development of a new tumor-bearing humanized mouse model to evaluate the efficacy and toxicity of bispecific T cell engagers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2875.

The effect of HTLV1 infection on inflammatory and oxidative parameters in the liver, kidney, and pancreases of BALB/c mice.

Viral infections are linked to the progression of inflammatory reactions and oxidative stress that play pivotal roles in systemic diseases. To confirm this phenomenon, in the present study, TNF‐α level and oxidative stress markers were examined in the liver, kidney, and pancreas of HTLV1‐infected male BALB/c mice. To this end, twenty BALB/c mice were divided into HTLV1‐infected mice that were inoculated with 1‐million HTLV1‐infected cells (MT‐2), and the control groups. Two months after inoculation, the peripheral blood, mesenteric lymph nodes, liver, kidney, and pancreas were collected after deep anesthetization of mice (ketamine, 30 mg/kg). The extracted DNA of mesenteric lymph nodes was obtained to quantify proviral load (PVL) using quantitative real‐time polymerase chain reaction (qRT‐PCR). The levels of lipid peroxidation, total thiol (SH), nitric oxide (NO), TNF‐α, catalase (CAT), and superoxide dismutase (SOD) activities were examined in the liver, kidney, and pancreases. Furthermore, histopathological changes in the liver and kidney were evaluated. In liver tissue, the levels of MDA, TNF‐α, and blood cell infiltration were significantly increased, and the levels of CAT and SOD were significantly decreased. In the kidney, a reduction in SOD, CAT, and total SH and an increase in MDA and NO were observed. In the pancreas, CAT activity, total SH, and SOD were decreased, and the levels of MDA and NO were enhanced. In terms of TNF‐α production, it has been shown that the level of this inflammatory cytokine was increased in the liver, kidney, and pancreas. The HTLV1 may have a role in inducing inflammatory reactions and oxidative stress pathways in the tissues.

Synthesis, Characterization, and Cytotoxicity Evaluation of Methotrexate-Polyethylene Glycol-Glutamic Acid Nanoconjugate as Targeted Drug Delivery System in Cancer Treatment

Introduction: Methotrexate (MTX) is used as a folic acid antagonist in the treatment of many human cancers. Attachment of hydrophilic ligands to MTX improves its efficacy due to reducing toxicity and enzymatic degradation and it also increases its in-vivo half-life. Materials and Methods: In the present study, pH-responsive nanoconjugates of methoxy poly(ethylene glycol)-glutamic acid methotrexate (mPEG-Glu-MTX) have been prepared and characterized using hydrogen nuclear magnetic resonance (1H-NMR) and Fourier transform infrared (FT-IR). Glutamic acid is attached to the mPEG chain by the carboxylic group and to the MTX via an amide bond to the amine group. Results: The prepared nanoconjugate has the mean diameter ranging from 160 to 190 nm and, the drug release was significantly induced two times at the pH of 5.5 and 3.5 compared with pH 7.4 (P &lt; 0.05). The prepared mPEG-Glu-MTX nanoconjugate showed toxicity similar to AGS, MDA, and MCF7 cell lines compared with the free form of MTX (P &gt; 0.1), which indicates that the conjugation does not effect on the MTX cytotoxicity but is expected to be successful in the targeted delivery of MTX. Conclusion: The results show that manufactured nanoconjugates can be considered as an efficient drug delivery system in the treatment of cancer; however, further studies are needed on the targeting activity of this nanocarrier in in-vivo conditions.

Heavy Metal Complexes of 1, 2, 3-Triazole derivative: Synthesis, Characterization, and Cytotoxicity Appraisal Against Breast Cancer Cell Lines (MDA-MB-231)

New chelating ligand derived from triazole and its complexes with metal ions Rhodium, Platinum and Gold were synthesized. Through a copper (I)-catalyzed click reaction, the ligand produced 1,3-dipolar cycloaddition between 2,6-bis((prop-2-yn-1-yloxy) methyl) pyridine and 1-azidododecane. All structures of these new compounds were rigorously characterized in the solid state using spectroscopic techniques like: 1HNMR, 13CNMR, Uv-Vis, FTIR, metal and elemental analyses, magnetic susceptibility and conductivity measurements at room temperature, it was found that the ligand acts as a penta and tetradentate chelate through N3O2, N2O2, and the geometry of the new complexes are identified as octahedral for (Rh &amp; Pt) complexes and for (Au) complex square planner. The newly prepared compounds were designed and efficiently synthesized to be used to investigation of their toxicity bioassay (in vitro) as anticancer agent towards MDA cell lines. From the results obtained from cytotoxic assay, it can be concluded that the synthesized compounds are promising as new anticancer candidates in future especially in high concentration.

Comparison of FTIR spectra of different breast cell lines to detect spectral biomarkers of pathology

Many efforts have been made for several years to make Fourier Transform Infrared (FTIR) micro-spectroscopy usable in the cytological diagnostic. In fact, some events arising when cells undergo a disease, as alterations in the relative content of biochemical cellular components and variations of the proteins secondary structure, lead to modifications of the FTIR spectra, as changes of relative intensity and/or spectral shifts of specific peaks. In this study, we compare FTIR spectra measured for three different breast cell lines: the non-malignant MCF10A cell line, the malignant non-metastatic MCF7 cell line and the metastatic MDA cell line. The major differences of average absorption spectra involve a blue-shift of amide I and II bands and a larger relative intensity of nucleic acid bands in MCF7 spectra with respect to the other ones. Curve fitting also provided important information about spectral markers able to discriminate the three cell lines. In fact, the ratio of sum of the areas of peaks at 2852 and 2922 cm−1 with respect to the sum of the areas of peaks at 2873 and 2958 cm−1 is able to differentiate the non-malignant, malignant and metastatic cells among them. These results suggest that FTIR micro-spectroscopy may be a potentially useful tool for breast cancer diagnosis at cytological level.

In vivo assessment of APPJ discharge on the earthworm: coelomic TAC and MDA levels, cell death, and tissue regeneration.

The effective medical applications of cold atmospheric pressure plasma jet (APPJ) have been reported by many researchers, including sterilization of liquid and solid surfaces, treatment of chronic wounds, treatment of cancer tumors, and blood clots. Therefore, in this study, we exposed Aporrectodea trapezoides and Eisenia fetida earthworms to APPJ discharge (0-30s) to evaluate the impacts on regeneration of missed segments, apoptosis, lipid peroxidation (LPo), catalase activity (CAT), total antioxidant capacity (TAC), total proteins, and protein profile. Results showed APPJ induced significant effects on regeneration ability of earthworms after 20 and 30s of exposure (p < 0.05). Atmospheric pressure plasma jet did not have significant effects on MDA content and TUNEL-positive cells, but this effect was significant for TAC and CAT in both species (p < 0.05). In conclusion, the present study revealed for the first time that regeneration of missed segments in earthworms can be stimulated by plasma treatment.

Identification of novel anti-cancer agents by the synthesis and cellular screening of a noscapine-based library

Noscapine is a natural product first isolated from the opium poppy (Papaver somniferum L.) with anticancer properties. In this work, we report the synthesis and cellular screening of a noscapine-based library. A library of novel noscapine derivatives was synthesized with modifications in the isoquinoline and phthalide scaffolds. The so generated library, consisting of fifty-seven derivatives of the natural product noscapine, was tested against MDA-MB-231 breast cancer cells in a cellular proliferation assay (with a Z’ > 0.7). The screening resulted in the identification of two novel noscapine derivatives as inhibitors of MDA cell growth with IC50 values of 5 µM and 1.5 µM, respectively. Both hit molecules have a five-fold and seventeen-fold higher potency, compared with that of lead compound noscapine (IC50 26 µM). The identified active derivatives retain the tubulin-binding ability of noscapine. Further testing of both hit molecules, alongside the natural product against additional cancer cell lines (HepG2, HeLa and PC3 cells) confirmed our initial findings. Both molecules have improved anti-proliferative properties when compared to the initial natural product, noscapine.