Comparison of the differentiation of ovine fetal bone-marrow mesenchymal stem cells towards osteocytes on chitosan/alginate/CuO-NPs and chitosan/alginate/FeO-NPs scaffolds.

In the current study, the creation of a chitosan/alginate scaffold hydrogel with and without FeO-NPs or CuO-NPs was studied. From fetal ovine bone marrow mesenchymal stem cells (BM-MSCs) were isolated and cultivated. Their differentiation into osteocyte and adipose cells was investigated. Also, on the scaffolds, cytotoxicity and apoptosis were studied. To investigate the differentiation, treatment groups include: (1) BM-MSCs were plated in DMEM culture medium with high glucose containing 10% FBS and antibiotics (negative control); (2) BM-MSCs were plated in osteogenic differentiation medium (positive control); (3) positive control group + FeO-NPs, (4) positive control group + CuO-NPs; (5) BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate scaffold; (6) BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate/FeO-NPs scaffold; and (7) BM-MSCs were plated in osteogenic differentiation medium on chitosan/alginate/CuO-NPs scaffold. Alkaline phosphatase enzyme concentrations, mineralization rate using a calcium kit, and mineralization measurement by alizarin staining quantification were evaluated after 21 days of culture. In addition, qRT-PCR was used to assess the expression of the ALP, ColA, and Runx2 genes. When compared to other treatment groups, the addition of CuO-NPs in the chitosan/alginate hydrogel significantly increased the expression of the ColA and Runx2 genes (p < 0.05). However, there was no significant difference between the chitosan/alginate hydrogel groups containing FeO-NPs and CuO-NPs in the expression of the ALP gene. It appears that the addition of nanoparticles, in particular CuO-NPs, has made the chitosan/alginate scaffold more effective in supporting osteocyte differentiation.

Evaluation of DNA damage induced by acesulfame potassium: spectroscopic, molecular modeling simulations and toxicity studies.

Acesulfame potassium (Ace-K) is a widely used artificial sweetener that has been reported to interact with DNA and cause important genetic damage. However, the type of interaction mechanism is unknown. This study provides an approach to understanding the in vitro mechanism of Ace-K interaction with Ct-DNA using spectroscopic methods combined with molecular simulations. The hypochromic effect as obtained from UV-Vis spectra indicated the formation of the DNA-Ace-K complex in the minor groove. Further evidence for groove binding mode comes from the decrease in Hoechst-DNA fluorescence caused by increasing Ace-K concentrations, alongside no detectable change in MB-DNA emission band intensity. A negative value of ΔH and ΔS represents the hydrogen bonds and van der Waals forces between Ace-K and DNA. Based on the molecular docking, Ace-K was located between the guanine10 and 16 in DNA minor groove and stabilized by two hydrogen bonds and one π-Sulfur interaction. In vitro cell culture results showed that about 5 mg/mL of Ace-K caused the death of 85% of HUVEC cells after 48 h. Communicated by Ramaswamy H. Sarma.

Inhibitory effects of postbiotic consisting sonication-killed Bifidobacterium bifidum on experimental triple negative breast neoplasm in mice: a preliminary study.

Background and Objectives: Breast cancer is the second leading cause of death and one of the most common malignancies among women in the world. The aim of this study was to investigate the preventive effects of postbiotic consisting of sonicated Bifidobacterium bifidum cells on triple negative breast cancer. Materials and Methods: Thirty-six female BALB/c mice aged 5-7 weeks were randomly divided into 3 groups (n=12): Ctrl-, healthy mice; Ctrl+, mice with breast cancer with no treatment; and Postbiotic, mice with orally gavage postbiotic before and after 4T1 cell line transplantation. Cancer progress and the effects of postbiotic were assayed by histological, immunohistochemical and gene expression quantification. Results: The histological results showed that administration postbiotic consisting of B. bifidum significantly decreased carcinogenesis in terms of tumor incidence, multiplicity and volume. The tumor progress was suppressed by oral intake of B. bifidum as showed by p53 and Ki-67 expression. Furthermore, Oral intake of postbiotic resulted in extended survival of mice and inhibited sever weigh loss. Conclusion: Pretreatment with sonication killed B. bifidum, as a postbiotic, inhibited breast cancer progress and malignancy.

The cell cycle, cancer development and therapy.

The process of cell division plays a vital role in cancer progression. Cell proliferation and error-free chromosomes segregation during mitosis are central events in life cycle. Mistakes during cell division generate changes in chromosome content and alter the balances of chromosomes number. Any defects in expression of TIF1 family proteins, SAC proteins network, mitotic checkpoint proteins involved in chromosome mis-segregation and cancer development. Here we discuss the function of organelles deal with the chromosome segregation machinery, proteins and correction mechanisms involved in the accurate chromosome segregation during mitosis.

Molecular and Biochemical Evaluation of Ethyl Methanesulfonate-Induced Mutant Lines in Camelina sativa L.

Background: Camelina sativa is one of the most important oilseeds that has a proportionate profile of essential unsaturated fatty acids that are suitable for human nutrition. In this regard, we can mention a high percentage and a reasonable ratio of omega 3 and omega 6. Objectives: In the current study, the created variation of second-generation mutant (M2) camelina lines in terms of fatty acid profiles and ISSR molecular markers in C. sativa was evaluated. Materials and Methods: For this purpose, while producing the first-generation of mutant plants (M1), 200 M2 seeds with 0.1% and 0.5% ethyl methanesulfonate (EMS) mutations were treated in two replications for 8 and 16 hours based on a completely randomized design. Results: The results of mean comparisons showed that there was no significant difference between treatments in terms of fatty acids of palmitic acid, stearic acid, linoleic acid, eicosadienoic acid, oleic acid and erucic acid. The cluster analysis revealed that all the treatments used with five replications were divided into eight groups. It was found that all replications of the treatment with a concentration of 0.1% and a time of 16 hours (C1T2) were in the second group with the lowest palmitic acid was present among other treatments. Therefore, C1T2 treatment is recommended as the best treatment to reduce palmitic acid. Examination of the information content of ISSR molecular markers also showed that markers 2, 5, and 6 were the best informative markers in the detection of camelina fatty acid profiles. Conclusion: A significant variation has been created in the fatty acids profile and it can be applied in future breeding programs depending on the intended purpose.

MUC1-Targeted Radiopharmaceuticals in Cancer Imaging and Therapy.

Mucin 1 (MUC1) is a large, transmembrane mucin glycoprotein overexpressed in most adenocarcinomas and plays an important role in tumor progression. Regarding its cellular distribution, biochemical features, and function, tumor-related MUC1 varies from the MUC1 expressed in normal cells. Therefore, targeting MUC1 for cancer immunotherapy and imaging can exploit the difference between cancerous and normal cells. Radiopharmaceuticals have a potential use as carriers for the delivery of radionuclides to tumors for a diagnostic imaging and radiotherapy. Several radiolabeled targeting molecules like peptides, antibodies, and aptamers have been efficiently demonstrated in detecting and treating cancer by targeting MUC1. This review provides a brief overview of the current status of developments and applications of MUC1-targeted radiopharmaceuticals in cancer imaging and therapy.

Key parameters optimization of chitosan production from Aspergillus terreus using apple waste extract as sole carbon source.

Chitosan is commonly obtained from shrimp and crab shell chitin by deacetylation; however, such supplies appear limitation. An alternative source of chitosan is cell wall in certain fungi. In this study, chitosan production through submerged fermentation of Aspergillus terreus on apple waste extract as sole carbon source was investigated. Monod equation with a maximum specific growth rate of 0.083 h-1 and half-saturation constant of 6.67 w/v% was best described the kinetic of growth. Results of response surface methodology showed the highest chitosan to substrate yield of 49.32 mg gsubstrate-1, chitosan to fungal biomass yield of 140.9 mg gcell-1, and fungal biomass to substrate yield of 0.387 gcell gsubstrate-1 were simultaneously obtained at temperature 30.0 °C, initial pH 5.98, and ammonium nitrate concentration 5.0 g L-1. The chitosan produced at the optimum condition was characterized by FTIR, TGA, and DSC analysis, and degree of deacetylation was 88.2%.

Nontoxic silver nanocluster-induced folding, fibrillation, and aggregation of blood plasma proteins.

In recent years, concerns have been raised considering the potential risks of nanocluster (NC) for the environment and human health. Since the blood circulation system is probably the first entry route of NC into the human body, adsorption of blood proteins on NC may change cellular responses, including cellular uptake efficiency, bio distribution patterns, and nanotoxicity profiles, besides other biological effects. Therefore, the interaction of NCs with proteins and the cellular implications can be therapeutically of great importance. Adsorption of human blood proteins on NCs has been methodically investigated. In the present study, the first analysis of fibrillation was conducted between MBI-AgNCs and human serum (a complex biofluid). AgNCs were prepared by coating with 2-mercaptobenzimidazole. Then, interactions with human blood proteins, such as immunoglobulin, albumin, and insulin were investigated using various experimental approaches. Upon protein association, the fluorescence of proteins significantly decreased, accompanied by a blue shift in the AgNCs-human serum albumin (HSA) system and a red shift in the AgNCs-insulin/γ-globulin. Concomitantly, circular dichroism spectroscopy and atomic force microscopy were employed to investigate the effects of protein binding to NCs. We found that AgNCs induced γ-globulin aggregation. HSA at the AgNC surface was partially unfolded and could promote protein self-assembly into amyloid fibrils, while the surface morphology remained unchanged after insulin incubation. The atomic force microscopy (AFM) data and the ThT and CR analysis of the proteins, as well as circular dichroism (CD) and fluorescence findings, support the use of AgNCs as an indicator for monitoring the progress of HSA fibrillogenesis. Additionally, cytotoxicity assays were used to ensure the biocompatibility of nanoparticles within the applicable limits.

Green synthesis and characterizations of gold nanoparticles using Thyme and survey cytotoxic effect, antibacterial and antioxidant potential.

Nowadays, green chemistry methods are noticeable for synthesis metal nanoparticles due to affordable, easy and high reaction rate. In the present study gold nanoparticles (Au NPs) were eco-friendly synthesized using Thyme extract at the room temperature for 30 min to provide non-toxic, which can be used for different applications. Identifying properties of synthesized gold nanoparticles was done by various analytical technique including UV-Vis absorption spectroscopy approved presence of Au NPs in the solution, the functional groups of Thyme extract in the reduction and capping process of Au NPs are determined by FT-IR, Crystalline with the fcc plane approved by X-ray diffraction (XRD) pattern, energy dispersive spectroscopy (EDS) determined existence of elements in the sample, surface morphology, diverse shapes and size of present Au NPs were showed by scanning electron microscopy (SEM), atomic force microscopy (AFM) and Transmission electron microscopy (TEM). Beginning and end destroy temperature of the gold nanoparticles were determined by thermal gravimetric spectroscopy (TGA). In addition, antibacterial, antioxidant, and cytotoxicity properties of Au NPs were studied. Antibacterial activity of Au NPs was investigated on gram-positive (Bacillus) and gram-negative (Escherichia coli) by disk diffusion; also MIC (Minimum Inhibitory Concentration) and MBC (Minimum Bactericidal Concentration) were determined. DPPH free radical scavenging assay was used for antioxidant property and compared to butylated hydroxy toluene (BHT) as a standard antioxidant that showed high antioxidant activity. Synthesized Au NPs have great cell viability in a dose-depended manner and demonstrate that this method provided nontoxic for synthesis gold nanoparticles. The average diameter of synthesized Au NPs was about 35 nm.

Fabrication of a novel enzymatic electrochemical biosensor for determination of tyrosine in some food samples.

In this work, fabrication of a novel and ultrasensitive electrochemical biosensor based on immobilization of tyrosine hydroxylase onto palladium-platinum bimetallic alloy nanoparticles/chitosan-1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl) imide/graphene-multiwalled carbon nanotubes-IL/glassy carbon electrode for determination of L-tyrosine in some high tyrosine foods including cheese, egg and yogurt was reported. Immobilization of tyrosine hydroxylase onto the surface of the biosensor was performed by cross-linking tyrosine hydroxylase and chitosan through the addition of glutaraldehyde. Enzymatic biosensors employ the affinity and selectivity of catalytically active proteins towards their target molecules and here, the tyrosine hydroxylase selectively catalyzes the conversion of tyrosine to levodopa which can be oxidized at lower potentials than tyrosine. The modifications were characterized by electrochemical impedance spectroscopy, cyclic voltammetry, energy dispersive X-ray spectroscopic and scanning electron microscopy. Under optimal conditions, the biosensor detected tyrosine in concentration ranges of 0.01 × 10-9 to 8.0 × 10-9 mol L-1 and 8.0 × 10-9 to 160.0 × 10-9 mol L-1 with a limit of detection of 0.009 × 10-9 mol L-1. The biosensor was able to selective determination of tyrosine even in the presence of common interferents therefore, the biosensor was highly selective. The biosensor also showed good operational stability, antifouling properties, sensitivity, repeatability and reproducibility.

Intellectual modifying a bare glassy carbon electrode to fabricate a novel and ultrasensitive electrochemical biosensor: Application to determination of acrylamide in food samples.

Acrylamide (AA) is a neurotoxin and carcinogen which is mainly formed in foods containing large quantities of starch processed at high temperatures and its determination is very important to control the quality of foods. In this work, a novel electrochemical biosensor based on hemoglobin-dimethyldioctadecylammonium bromide (HG-DDAB)/platinum-gold-palladium three metallic alloy nanoparticles (PtAuPd NPs)/chitosan-1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (Ch-IL)/multiwalled carbon nanotubes-IL (MWCNTs-IL)/glassy carbon electrode (GCE) is proposed for ultrasensitive determination of AA in food samples. Development of the biosensor is based on forming an adduct by the reaction of AA with α-NH2 group of N-terminal valine of HG which decreases the peak current of HG-Fe+3 reduction. The modifications were characterized by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), energy dispersive X-ray spectroscopic (EDS) and scanning electron microscopy (SEM). Under optimized conditions, the biosensor detected AA by square wave voltammetry (SWV) in two linear concentration ranges of 0.03-39.0nM and 39.0-150.0nM with a limit of detection (LOD) of 0.01nM. The biosensor was able to selective detection of AA even in the presence of high concentrations of common interferents which confirmed that the biosensor is highly selective. Also, the results obtained from further studies confirmed that the proposed biosensor has a short response time (less than 8s), good sensitivity, long term stability, repeatability, and reproducibility. Finally, the proposed biosensor was successfully applied to determine AA in potato chips and its results were comparable to those obtained by gas chromatography-mass spectrometry (GC-MS) as reference method.

Radioprotective effects of lentil sprouts against X-ray radiation.

The present study investigated the radioprotective efficacy of lentil (Lens culinaris) sprouts against X-ray radiation-induced cellular damage. Lentil seeds were dark germinated at low temperature and the sprout extract was prepared in PBS. Free radical scavenging of extract was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and then the radioprotective potency of extract (0 to 1000 µg/mL) on the lymphocyte cells was determined by lactate dehydrogenases assay. Moreover, micronuclei assay was assessed using the cytokinesis-block technique. The irradiations were performed using 6 MV X-ray beam. The value of IC50 for DPPH assay was 250 µg/mL. The median lethal dose for radiation was determinate at 5.37 Gy. Pretreatment with lentil sprout extract at 1000 µg/mL reduced cytotoxicity at 6 Gy total concentration from 70% to 50%. The results of micronuclei assay indicated that cells were resistant to radiation at concentrations of 500-1000 µg/mL of exogenous lentil sprout extract. The value of median effective concentration for micronuclei assay was 500 µg/mL. The results indicated that lentil sprout extract showed actually somewhat radioprotective effect on lymphocyte cell. In addition, the obtained results suggest that extract of total lentil sprout have more antioxidant activity than radicle part.

An improved radiolabelled RNA aptamer molecule for HER2 imaging in cancers.

Human epidermal growth factor receptor 2 (HER2) expression has been shown to be increased in several types of human tumours. In this study, for the imaging of HER2-related tumours, a modified RNA aptamer with HER2-specific targeting was labelled with (99m)Tc, by using hydrazino nicotinamide (HYNIC) as the chelator in the presence of tricine or ethylenediamine-N,N'-diacetic acid (EDDA) as the co-ligand. Stability testing of the radiolabelled aptamers in the serum was performed through SDS-PAGE. The aptamer-radionuclide conjugate was evaluated for its cellular HER2-specific binding in ovarian cancer cells (SKOV-3), and its biodistribution properties were assessed in normal and SKOV-3 tumour-bearing mice. In the presence of either tricine or EDDA, the HYNIC-RNA aptamers were labelled with (99m)Tc at a high yield and radiochemical purity. Cellular experiments confirmed the specific binding of the RNA aptamer to the HER2 receptor. In the animal biodistribution study, uptake of the EDDA-co-liganded (99m)Tc-HYNIC-RNA aptamer by the liver and spleen was remarkably lower than that of the aptamer with tricine. Tumours also showed a higher accumulation of radioactivity with the EDDA-co-liganded aptamer complex. This study demonstrated EDDA to be better than tricine for use as a co-ligand with the RNA aptamer, which can be a potential tool for the molecular imaging of HER2-overexpressing cancers.

A HER2-targeted RNA aptamer molecule labeled with 99mTc for single-photon imaging in malignant tumors.

A modified RNA aptamer with HER2-specific binding was conjugated to hynic and labeled with (99m)Tc, for potential use as a radiopharmaceutical for diagnostic imaging of ovarian cancer cells (SKOV-3) with high HER2 expression. The aptamer was radiolabeled with (99m)Tc by using hynic as the chelator and tricine as the co-ligand. Stability testing of the radioconjugated aptamer was performed via ITLC and SDS-PAGE in normal saline and serum. The aptamer-radionuclide conjugate was evaluated for cellular HER2-specific binding, saturation affinity, and cellular internalization in SKOV-3 and MCF-7 cells, and its biodistribution properties were assessed in normal and SKOV-3 tumor-bearing mice. Radiolabeling of the aptamer was achieved with high yield and radiochemical purity, and the (99m)Tc-hynic-RNA aptamer was highly stable in normal saline and serum. Cellular experiments showed specific binding of the aptamer to the HER2 receptor with a dissociation constant of 27 nM. Rapid blood clearance was observed after injection of the (99m)Tc-hynic-RNA aptamer, and the main excretion route was via the hepatobilary system. While the radioconjugated aptamer bound specifically to the HER2 receptor on cells in vitro, it did not show any significant tumor-to-blood or tumor-to-muscle ratios in mice. Modifications to radiolabeled aptamer will require improving its pharmacokinetic properties and tumor uptake in vivo.

Radioimmunotherapy with ¹³¹I-bevacizumab as a specific molecule for cells with overexpression of the vascular endothelial growth factor.

Bevacizumab is a humanized monoclonal antibody that inhibits vascular endothelial growth factor A and is used for the treatment of several cancers. We labeled this monoclonal antibody with Iodine-131 (¹³¹I) and performed in vitro quality control and tumor cell growth inhibition tests. Bevacizumab was labeled with ¹³¹I using chloramine T. Radiochemical purity and stability in phosphate-buffered saline and human blood serum were determined using thin-layer chromatography and radio-sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively, performed at different times. Cell-specific binding, internalization, and toxicity of the radiolabeled antibody were tested using the SKOV-3 ovarian cancer cell line. The biodistribution of ¹³¹I-bevacizumab was investigated using male mice. The radiochemical purity of the complex was 99% ± 0.7%. Its stability in phosphate-buffered saline and human blood serum at 48 hours postpreparation was 78% ± 1.2% and 93% ± 0.6%, respectively. (131)I-bevacizumab was significantly bound to SKOV-3. The internalization of ¹³¹I-bevacizumab was time dependent, and it was cleared from the blood after 24 hours. Significant reductions in SKOV-3 cell viability were achieved with (131)I-bevacizumab at a concentration of 500 nM. A low accumulation of ¹³¹I-bevacizumab was observed in the stomach and salivary glands after 24 hours and 48 hours. These findings indicate that the new radiolabeled antibody should be further evaluated in animals and, possibly, in humans as a new radiopharmaceutical agent for use in radioimmunotherapy for ovarian cancer.