TGM2, HMGA2, FXYD3, and LGALS4 genes as biomarkers in acquired oxaliplatin resistance of human colorectal cancer: A systems biology approach.

Acquired resistance to oxaliplatin is considered as the primary reason for failure in colorectal cancer (CRC) therapy. Identifying the underlying resistance mechanisms may improve CRC treatment. The present study aims to identify the key genes involved in acquired oxaliplatin-resistant in CRC by confirming the oxaliplatin resistance index (OX-RI). To this aim, two public microarray datasets regarding oxaliplatin-resistant CRC cells with different OX-RI, GSE42387, and GSE76092 were downloaded from GEO database to identify differentially expressed genes (DEGs). The results indicated that the OX-RI affects the gene expression pattern significantly. Then, 54 common DEGs in both datasets including 18 up- and 36 down-regulated genes were identified. Protein-protein interaction (PPI) analysis revealed 13 up- (MAGEA6, TGM2, MAGEA4, SCHIP1, ECI2, CD33, AKAP12, MAGEA12, CALD1, WFDC2, VSNL1, HMGA2, and MAGEA2B) and 12 down-regulated (PDZK1IP1, FXYD3, ALDH2, CEACAM6, QPRT, GRB10, TM4SF4, LGALS4, ALDH3A1, USH1C, KCNE3, and CA12) hub genes. In the next step, two novel up-regulated hub genes including ECI2 and SCHIP1 were identified to be related to oxaliplatin resistance. Functional enrichment and pathway analysis indicated that metabolic pathways, proliferation, and epithelial-mesenchymal transition may play dominant roles in CRC progression and oxaliplatin resistance. In the next procedure, two in vitro oxaliplatin-resistant sub-lines including HCT116/OX-R4.3 and HCT116/OX-R10 cells with OX-IR 3.93 and 10.06 were established, respectively. The results indicated the up-regulation of TGM2 and HMGA2 in HCT116/OX-R10 cells with high OX-RI and down-regulation of FXYD3, LGALS4, and ECI2 in both cell types. Based on the results, TGM2, HMGA2, FXYD3, and LGALS4 genes are related to oxaliplatin-resistant CRC and may serve as novel therapeutic targets.

The effect of ciprofloxacin on doxorubicin cytotoxic activity in the acquired resistance to doxorubicin in DU145 prostate carcinoma cells.

The present study aimed to assess the influence of ciprofloxacin (CIP) against the doxorubicin (DOX)-resistant androgen-independent prostate cancer DU145 cells. The DOX-resistant DU145 (DU145/DOX20) cells were established by exposing DU145 cells to the increasing concentrations of DOX. The antiproliferative effect of CIP was examined through employing MTT, colony formation, and 3D culture assays. DU145/DOX20 cells exhibited a twofold higher IC50 value for DOX, an increased ABCB1 transporter activity, and some morphological changes accompanied by a decrease in spheroid size, adhesive and migration potential compared to DU145 cells. CIP (5 and 25 µg mL-1) resulted in a higher reduction in the viability of DU145/DOX20 cells than in DU145 cells. DU145/DOX20 cells were more resistant to CIP in 3D culture compared to the 2D one. No spheroid formation was observed for DU145/DOX20 cells treated with DOX and CIP combination. CIP and DOX, alone or in combination, significantly reduced the growth of DU145 spheroids. CIP in combination with 20 nM DOX prevented the colony formation of DU145 cells. The clonogenicity of DU145/DOX20 cells could not be estimated due to their low adhesive potential. CIP alone caused a significant reduction in the migration of DU145 cells and resulted in a more severe decrease in the wound closure ability of DOX-exposed ones. We identified that CIP enhanced DOX sensitivity in DU145 and DU145/DOX20 cells. This study suggested the co-delivery of low concentrations of CIP and DOX may be a promising strategy in treating the DOX-resistant and -sensitive hormone-refractory prostate cancer.

Differential expression analysis of genes and long non-coding RNAs associated with KRAS mutation in colorectal cancer cells.

KRAS mutation is responsible for 40-50% of colorectal cancers (CRCs). RNA-seq data and bioinformatics methods were used to analyze the transcriptional profiles of KRAS mutant (mtKRAS) in comparison with the wild-type (wtKRAS) cell lines, followed by in-silico and quantitative real-time PCR (qPCR) validations. Gene set enrichment analysis showed overrepresentation of KRAS signaling as an oncogenic signature in mtKRAS. Gene ontology and pathway analyses on 600 differentially-expressed genes (DEGs) indicated their major involvement in the cancer-associated signal transduction pathways. Significant hub genes were identified through analyzing PPI network, with the highest node degree for PTPRC. The evaluation of the interaction between co-expressed DEGs and lncRNAs revealed 12 differentially-expressed lncRNAs which potentially regulate the genes majorly enriched in Rap1 and RAS signaling pathways. The results of the qPCR showed the overexpression of PPARG and PTGS2, and downregulation of PTPRC in mtKRAS cells compared to the wtKRAS one, which confirming the outputs of RNA-seq analysis. Further, significant upregualtion of miR-23b was observed in wtKRAS cells. The comparison between the expression level of hub genes and TFs with expression data of CRC tissue samples deposited in TCGA databank confirmed them as distinct biomarkers for the discrimination of normal and tumor patient samples. Survival analysis revealed the significant prognostic value for some of the hub genes, TFs, and lncRNAs. The results of the present study can extend the vision on the molecular mechanisms involved in KRAS-driven CRC pathogenesis.

Zinc oxide nanofluids: The influence of modality combinations on prostate cancer DU145 cells.

AIM: The combination of phototherapy and chemotherapy (chemophototherapy), presents a promising multimodal method for comprehensive cancer treatment. The aim of this study is to investigate the influence of low doses of zinc oxide (ZnO) nanofluids and ultraviolet A (UVA) irradiation on the cytotoxicity and cellular uptake of doxorubicin (DOX) on human prostate cancer DU145 cells. MATERIALS AND METHODS: ZnO nanoparticles were prepared by the solvothermal method and 10% bovine serum albumin was used as the dispersant. The cytotoxic effect of DOX alone and in combination with different concentrations of ZnO nanofluids (0.95-15.6 µg/ml) in the presence and absence of UVA irradiation on DU145 cells was evaluated by -(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. DOX residue inside and outside of DU145 cells was explored by fluorescence microscopy and UV-Vis absorption spectroscopy, respectively. The role of ZnO nanofluids and UVA irradiation in DOX-induced apoptosis and cell cycle arrest were evaluated by DAPI staining, comet assay, and flow cytometry. RESULTS: The results revealed that low dose of ZnO nanofluids (0.95 µg/ml) accompanied with irradiation enhanced the cytotoxicity and intracellular delivery of DOX in DU145 cells. The percentage of chromatin fragmentation/condensation and DNA tail of DU145 cells treated simultaneously with DOX and ZnO nanofluids was increased after UVA irradiation, whereas no significant changes in cell cycle progression were observed. CONCLUSION: The results indicate that ZnO nanofluids in the presence of UVA irradiation could increase DOX efficiency in DU145 cells, suggesting such modality combinations as a promising approach in cancer treatment.

Kinetic and thermodynamic insights into the interaction of Aß1-42 with astaxanthin and aggregation behavior of Aß1-42: Surface plasmon resonance, microscopic, and molecular docking studies.

Amyloid-ß 1-42 (Aß1-42) aggregation is considered as an important process in the pathology of Alzheimer's disease (AD). Astaxanthin (ATX), a xanthophyll carotenoid, has a broad range of biological activities such as neuroprotective one. The present study aimed to elucidate the interaction of ATX with Aß1-42, as well as its effect on Aß1-42 aggregates under different conditions. Based on the surface plasmon resonance (SPR) results, ATX possessed a high affinity towards Aß1-42 and the binding process was spontaneous, endothermic, and entropy-driven. Additionally, the binding affinity of ATX to Aß1-42 was glucose and insulin concentration-dependent. Hydrophobic interactions may play an important role in the interaction between ATX and Aß1-42. The results of SPR, thioflavin T (ThT), and transmission electron microscopy (TEM) analyses represented the dependency of the anti-amyloid activity of ATX on glucose, insulin, and ATX concentrations. Further, molecular docking results indicated the presence of some same binding sites on Aß1-42 for ATX and glucose, as well as ATX and insulin, which suggests the possible competition between the molecules for Aß1-42 binding. Furthermore, the MTT results confirmed that ATX effect on the viability of Aß1-42-treated PC12 cells was dependent on glucose, insulin, and ATX concentrations. In general, the results provided further insights into the interaction between Aß1-42 and ATX, as well as the effect of ATX on Aß1-42 aggregates under various conditions.

Development and evaluation of a novel beneficent antimicrobial bioscaffold based on animal waste-fish swim bladder (FSB) doped with silver nanoparticles.

Treated fish wastes have found many applications in industry and medicine. Besides, nowadays low-cost scaffold with antimicrobial activity which can accelerates the process of wound healing is very demanding. In this study fish swim bladder (FSB), taken from Rutilus frisii, which is a disposable waste was doped with silver nanoparticles (AgNPs) and evaluated as antimicrobial wound dressing. The scanning electron microscopy (SEM) micrographs showed the presence of AgNPs on the scaffold. Histological observation confirmed cells and muscle removal from FSB and collagen preservation. There was significant antibacterial activity even in 50 ppm AgNPs concentration against pathogenic bacteria, swelling ratio was rather low, and cytotoxic assay revealed that the AgNPs-FSB scaffold had no toxic effect on human foreskin fibroblast (HFF) cells. Interestingly, despite the porous structure, the AgNPs-FSB scaffold was found to be a suitable barrier to microbial penetration even after 72 h. Further study showed the gradual release of AgNPs during 24 h. In conclusion, biofabricated FSB prepared in this study have appropriate characteristics notably encompassing a high quantity of collagen and broad-spectrum antimicrobial activity. Also, its porous structure made it suitable as a 3-D structure for the growth of cells and adding other antimicrobial nano-sized materials.

From basic researches to new achievements in therapeutic strategies of KRAS-driven cancers.

Among the numerous oncogenes involved in human cancers, KRAS represents the most studied and best characterized cancer-related genes. Several therapeutic strategies targeting oncogenic KRAS (KRAS onc ) signaling pathways have been suggested, including the inhibition of synthetic lethal interactions, direct inhibition of KRAS onc itself, blockade of downstream KRAS onc effectors, prevention of post-translational KRAS onc modifications, inhibition of the induced stem cell-like program, targeting of metabolic peculiarities, stimulation of the immune system, inhibition of inflammation, blockade of upstream signaling pathways, targeted RNA replacement, and oncogene-induced senescence. Despite intensive and continuous efforts, KRAS onc remains an elusive target for cancer therapy. To highlight the progress to date, this review covers a collection of studies on therapeutic strategies for KRAS published from 1995 to date. An overview of the path of progress from earlier to more recent insights highlight novel opportunities for clinical development towards KRASonc-signaling targeted therapeutics.

Identifying miltefosine-resistant key genes in protein-protein interactions network and experimental verification in Iranian Leishmania major.

Drug resistance is a complex phenomenon during leishmaniasis chemotherapy. In this study, the genes and pathways involved in miltefosine (MIL)-resistant Leishmania were identified using microarray data and in silico approaches. GSE30685 and GSE45496 were obtained from GEO database and analyzed with GEO2R tool to identify genes involved in MIL-resistant Leishmania. 177 differentially expressed genes (DEGs) were selected from these GSEs, which about half of them were uncharacterized/hypothetical proteins. The interactions between DEGs were investigated using STRING database and protein-protein interaction (PPI) networks. Five hub nodes were found in the PPI network. The gene ontology (GO) analysis of the resulting network revealed that DNA replication (GO:0006260) and ATP hydrolysis coupled proton transport (GO:0015991) were the most enriched GO term. Iranian MIL-resistant Leishmania major (L. major) parasites were generated by exposure of wild-type isolates to the increasing concentrations of MIL over a period of 5 months. Proof of mRNA expression levels of the obtained hub genes was assessed in Iranian wild-type and acquired resistant L. major parasites by real-time PCR. A significant higher expression level of LDBPK_150170 (encoding protein phosphatase 2C, PP2C), was only observed in Iranian L. major parasites resistance to MIL. Moreover, the RT-PCR results showed that the expression of metacyclic marker (small hydrophilic endoplasmic reticulum-associated protein, SHERP) and MIL-resistant marker (Leishmania MIL-transporter, LMT) was significantly increased and decreased, respectively, in Iranian MIL-resistant L. major parasites. Taken together, these data suggested that PP2C as well as SHERP and LMT genes may be prospective targets for the treatment of MIL-resistant Leishmania.

Concentration-Dependent Dual Effects of Ciprofloxacin on SB-590885-Resistant BRAFV600E A375 Melanoma Cells.

BRAF inhibitors (BRAFi) have been applied to treat melanoma harboring V600E mutations. Several studies showed that BRAFi-resistant melanomas are dependent on mitochondrial biogenesis. Therefore, the present study aimed to investigate the influence of ciprofloxacin (CIP), a mitochondria-targeting antibiotic, on SB-590885-resistant BRAFV600E A375 melanoma (A375/SB) cells. The cytotoxicity activity of CIP and SB-590885, a potent and specific BRAFi, on A375 and A375/SB cells was evaluated by MTT, colony formation, migration, and spheroid formation assays. Moreover, SB-590885-induced cell death in A375 cells was analyzed. SB-590885 showed time- and concentration-dependent cytotoxic effects on A375 cells. Twenty-five µg/mL CIP decreased the cell viability of A375 and A375/SB cells in a time-dependent manner. This concentration of CIP markedly decreased clonogenicity in both cells and caused a reduction in the growth of A375/SB spheroids. The cytotoxicity of 5 µg/mL CIP on A375/SB cells was less than that of A375 cells. The colony formation and migration ability of A375/SB cells was increased in the presence of 5 µg/mL CIP. Ten µM SB-590885 induced a massive vacuolization in A375 cells. Cell death assays suggested a simultaneous activation of autophagy, paraptosis, apoptosis, and necrosis. For the first time, this study reveals that CIP at the maximum concentration in serum (5 µg/mL) can enhance the colony formation and migration abilities in BRAFi-resistant melanoma cells, while it has cytotoxic activity against these cells at a higher concentration than serum level. This study suggests that CIP may promote aggressive growth properties in BRAFi-resistant melanomas, at a concentration present in serum.

Efficacy of metformin in mediating cellular uptake and inducing apoptosis activity of doxorubicin.

The clinical use of doxorubicin (DOX) is limited due to its systemic side effects and drug resistance. Recent evidence suggests that metformin prevents and controls certain but not all types of cancer. The beneficial use of metformin in combination with some chemotherapeutic agents has been reported. The aim of this study is to investigate the influence of metformin on DOX-induced effects in human prostate DU145 cancer cells and clarify its molecular mechanisms. For this purpose, DU145 cells were treated with DOX or metformin, either alone or in combination with each other. The proliferation of DU145 cells was inhibited by DOX-alone and metformin-alone treatment in a time and dose-dependent manner. Metformin could enhance the cytotoxicity of DOX by increasing DOX cellular uptake and cell cycle arrest at G1/S checkpoint which is associated with the enhancement of p21 protein expression. Moreover, metformin could elevate DOX-induced apoptosis in DU145 cells in a concentration-dependent manner and DOX-induced caspase-3 activity. These findings suggest that the combined treatment of metformin with DOX potentiates the anticancer efficacy of DOX in DU145 cells via inhibiting ABCB1 function, cell cycle arrest at G1/S transition and apoptosis induction.

In vitro antibacterial activity of ceftazidime, unlike ciprofloxacin, improves in the presence of ZnO nanofluids under acidic conditions.

The development of antibiotic resistance among hospital pathogens has provided a great need for new antimicrobial agents. Zinc oxide nanoparticles (ZnO NPs) in combination with various antibiotics can act as a reducing agent for antibiotic resistance. The aim of this study was to investigate the influence and the mechanism of ZnO NPs on the antimicrobial activity of ciprofloxacin (CP) and ceftazidime (CAZ) against Enterococcus faecalis (E. faecalis) and Acinetobacter baumannii (A. baumannii) bacteria in acidic conditions (pH 5.5). ZnO NPs were synthesised using the solvothermal method and characterised. The MIC90 value of ZnO NPs against A. baumannii was 0.25 mg ml-1 and its highest growth-inhibitory activity was observed at 0.125 mg ml-1 for E. faecalis. The Fourier transform infrared spectroscopy spectra of ZnO NPs treated with antibiotics showed the interaction between ZnO NPs and each of the two antibiotics. ZnO NPs at a sub-inhibitory concentration had no effect on the antibacterial activity of CP and CAZ against E. faecalis and CP against A. baumannii. The action mechanism of ZnO NPs for enhancing the antibacterial efficacy of CAZ against A. baumannii was evaluated. ZnO NPs caused to increase in the antibacterial activity of CAZ against A. baumannii, possibly through the release of Zn2+ and increasing of membrane permeability.

Poly-L-arginine: Enhancing Cytotoxicity and Cellular Uptake of Doxorubicin and Necrotic Cell Death.

OBJECTIVE: Cell resistance to doxorubicin and its toxicity to healthy tissue reduce its efficiency. The use of cell-penetrating peptides as drug delivery system along with doxorubicin is a strategy to reduce its side effects. In this study, the influence of poly-L-arginine on doxorubicin cytotoxicity, its cellular uptake and doxorubicin-induced apoptosis on human prostate cancer DU145 cells are assessed. METHODS: The cytotoxicity of doxorubicin and poly-L-arginine, alone and in combination, in DU145 cells was evaluated at different exposure times using MTT assay. The influence of poly-L-arginine on doxorubicin delivery into cells was evaluated by fluorescence microscopy and ultraviolet spectroscopy. DAPI and ethidium bromide- acridine orange stainings, flow cytometry using annexin V/propidium iodide, western blot analysis with anti-p21 antibody and caspase-3 activity were used to examine the influence of poly-L-arginine on doxorubicininduced cell death. RESULTS: Poly-L-arginine had no cytotoxicity at low concentrations and short exposure times. Poly-L-arginine increased the cytotoxic effect of doxorubicin in DU145 cells in a time-dependent manner. But no significant reduction was found in HFF cell viability. Poly-L-arginine seems to facilitate doxorubicin uptake and increase its intracellular concentration. 24h combined treatment of cells with doxorubicin (0.5 µM) and poly-L-arginine (1 µg ml-1) caused a small increase in doxorubicin-induced apoptosis and significantly elevated necrosis in DU145 cells as compared to each agent alone. CONCLUSION: Our results indicate that poly-L-arginine at lowest and highest concentrations act as proliferationinducing and antiproliferative agents, respectively. Between these concentrations, poly-L-arginine increases the cellular uptake of doxorubicin and its cytotoxicity through induction of necrosis.

Synthesis and antiproliferative and apoptosis-inducing activity of novel 3-substituted-3-hydroxy-2-oxindole compounds.

Anticancer role of oxindole compounds is well documented. Here, we synthesized new derivatives of 3-hydroxy-2-oxindole functionalized at position 3 (1a-f) which are expected to have antiproliferative activity in cancer cells. Human prostate cancer cell line (DU145) was treated with the synthesized derivatives at 40-µM concentration for 24, 48, and 72 h. Compounds 1-ethyl-3-hydroxy-1,1',3,3'-tetrahydro-2H,2'H-3,3'-biindole-2,2'-dione (1d), 5-bromo-1-ethyl-3-hydroxy-1,1',3,3'-2H,2'H-3,3'-biindole-2,2'-dione (1e), and 5-chloro-1-ethyl-3-hydroxy-1,1',3,3'-tetrahydro-2H,2'H-3,3'-biindole-2,2'-dione (1f) were found to significantly reduce DU145 cell viability at 48 and 72 h whereas no significant changes were observed up to 24 h. The compounds 1e and 1f showed the most cytotoxicity effect and had a similar antiproliferative activity on DU145 cell line. They have halogen and ethyl substitutions at positions 5 and 1, respectively. The IC50 of compound 1e for DU145 and A375 cells at 48 h was determined. The apoptotic effects and cell cycle progression of compound 1e at 1/2 × IC50 (55 µM) concentration in DU145 cells were investigated by nuclei staining, comet assay, flow cytometry, and scanning electron microscopy (SEM). The results obtained showed that this compound increased the percentage of tail DNA, increased the occurrence of the sub-G1 phase, and induced G2M arrest and apoptosis in DU145 cells after exposure for 48 h to a 55-µM concentration. The SEM images revealed cell contraction at 24 h, cell condensation, plasma membrane blebbing, and formation of apoptotic bodies at 48 and 72 h. These observations suggest that the antiproliferative activity of compound 1e may be to induce apoptosis in DU145 cells.

Antibacterial activity of poly-l-arginine under different conditions.

BACKGROUND AND OBJECTIVES: Arginine-rich peptides are an important class of antimicrobial peptides (AMPs) that exert their antibacterial activity via a lytic mechanism. Although the antibacterial activity of arginine-rich peptides has been already evaluated, no reports have so far been evaluated the influence of reaction conditions on their antimicrobial potential. The aim of the present study was to investigate the influence of pH, temperature, and glycine on antibacterial activity of poly-l-arginine (PLA) with a molecular weight of 5-15 kDa against Escherichia coli O157:H7 and Staphylococcus aureus. MATERIALS AND METHODS: The percentage of growth inhibition of PLA against both bacteria was analyzed at various pH, temperatures and sub-inhibitory concentrations of glycine by two-fold broth microdilution method. RESULTS: The results showed that PLA had antibacterial activity against E. coli O157:H7 and S. aureus and the inhibitory effect increased with increasing PLA concentration. The antimicrobial activity of PLA against both microorganisms was higher in basic media than under acidic or neutral conditions. At 1/2 times the MIC, heat treatment intensified the toxicity of PLA against E. coli O157:H7 whereas the susceptibility to PLA seems to be temperature independent for S. aureus. The MICs of glycine against E. coli O157:H7 and S. aureus were 12.5 and 25 mg ml-1, respectively. The antibacterial activity of PLA against both microorganisms increased, as indicated by the increasing growth inhibition percentage of this peptide with increasing glycine concentration. CONCLUSION: The antibacterial activity of PLA against S. aureus and E. coli O157:H7 depends on pH and glycine concentration.

Modulation of antibiotic resistance in Pseudomonas aeruginosa by ZnO nanoparticles.

BACKGROUND AND OBJECTIVES: Bacterial resistance to conventional antibiotics has become a widespread public health problem. The aim of this study was to investigate the influence of zinc oxide nanoparticles (ZnO NPs) on the antibacterial activity of several conventional antibiotics against Pseudomonas aeruginosa. MATERIALS AND METHODS: ZnO NPs were prepared by solvothermal method and dispersed in glycerol with the help of ammonium citrate as a dispersant. The antibacterial effects of the resulting ZnO nanofluid, ceftazidime, tobramycin, and ciprofloxacin were investigated against two P. aeruginosa strains, including one clinical isolate and P. aeruginosa ATCC 9027 using microdilution method. For the evaluation of the combined effect of ZnO nanofluid and antibiotics, the fractional inhibitory concentration indices were calculated and isobolograms were plotted. RESULTS: Clinical strain in comparison to standard strain of P. aeruginosa showed more resistance to ZnO nanofluid and the antibiotics. ZnO nanofluid acted synergistically with ceftazidime and tobramycin against both strains. Combination of ZnO nanofluid and ciprofloxacin displayed synergistic and partial synergistic activity against clinical and standard strains of P. aeruginosa, respectively. CONCLUSION: The results suggest that bacterial resistance to antimicrobials could be reduced by the synergistic action of ZnO NPs.

Direct and indirect sonication affect differently the microstructure and the morphology of ZnO nanoparticles: Optical behavior and its antibacterial activity.

In the present study, the sono-synthesis of ZnO nanoparticles (NPs) was performed by simple, low-cost, and the environmentally friendly method. The synthesis of zinc oxide as an antibacterial agent was performed by an ultrasonic bath (low intensity) for the indirect sonication and a horn system (high intensity) for the direct sonication. The samples synthesized by these two kinds of sonication were compared with each other. Crystallographic structures and the morphologies of the resultant powders were determined by X-ray diffraction (XRD) and transmission electron microscopy (TEM). The XRD patterns showed that both ZnO samples were crystallized in their pure phase. The TEM images confirmed that the morphologies of the products were completely different from each other. Based on the obtained analysis, the probable growth mechanisms were proposed for crystallization of both samples. The antibacterial activity of the synthesized species was evaluated by the colony count method against Escherichia coli O157:H7. Moreover, the optical behavior of the samples was studied by UV-vis spectroscopy and the variation of the ZnO band gap was compared.

Association of rs12255372 (TCF7L2) and D76N (PDX-1) Polymorphisms with Type 2 Diabetes in a Population Living in Northeast Iran.

The aim of this study was to investigate whether the rs12255372 (TCF7L2) and D76N (PDX-1) polymorphisms are associated with type 2 diabetes mellitus (T2DM) in Mashhad, northeast Iran. A hundred twenty seven patients with T2DM and 71 non-diabetic controls in Mashhad were genotyped by PCR-RFLP and ARMS-PCR methods. Single nucleotide polymorphisms (SNPs) were confirmed by sequencing in some samples and allelic and genotypic frequencies were then analyzed in each group. The T-allele of the SNP rs12255372 of TCF7L2 (OR = 2.70, 95% CI = 1.12-6.49, P = 0.027) and the A-allele of PDX-1 D76N (OR = 3.93, 95% CI = 1.60-7.68, P = 0.002) were significantly associated with an increased risk of T2DM. The rs12255372 SNP of TCF7L2 and D76N of PDX-1 genes may confer susceptibility to T2DM in the population living in Mashhad.

Effects of pH and Temperature on Antibacterial Activity of Zinc Oxide Nanofluid Against Escherichia coli O157: H7 and Staphylococcus aureus.

BACKGROUND: Zinc oxide nanoparticles (ZnO NPs) are known as one of the important inorganic materials used in research and health-related applications with effective antibacterial activities. Although the toxic effects of ZnO NPs have already been evaluated, more information is required to understand the possible mechanisms. OBJECTIVES: The aim of the present study was to determine the influences of pH and temperature on antibacterial activity of ZnO NPs against some strains of pathogenic bacteria. Identifying the interrelationship between toxicity and cultural conditions helps us to have a better understanding of the optimum reaction conditions for maximum antimicrobial activity. MATERIALS AND METHODS: ZnO NPs were prepared and characterized and then dispersed in glycerol with the help of ammonium citrate as the dispersant. The antibacterial tests were performed by measuring the growth of Escherichia coli O157:H7 and Staphylococcus aureus with different concentrations of ZnO NPs in glycerol. All the experiments were conducted at different incubation temperatures (25-42(°)C) and pH levels (4-10 for E. coli O157:H7 and 5-10 for S. aureus). RESULTS: The results showed that ZnO nanofluid have antibacterial activity against E. coli O157:H7 and S. aureus and the inhibitory effect increases with increasing the nanofluid concentration. The experiments showed that the antibacterial activity of ZnO NPs was influenced by temperature and pH. Higher antibacterial activity was observed at acidic pH levels with the maximum toxicity at pH = 4 and pH = 5 for E. coli O157: H7 and S. aureus, respectively. By raising the temperature, the toxicity of ZnO nanofluid increased, with the highest antibacterial activity at 42°C for both bacterial types in comparison with positive controls under the same conditions. CONCLUSIONS: Analysis of the results demonstrated that exposure media of ZnO NPs and cultural factors play a role in their cytotoxic effects. It could be attributed to the principal mechanism at different reaction conditions.

Spectroscopic and molecular modeling based approaches to study on the binding behavior of DNA with a copper (II) complex.

Blocking the division of tumor cells by small-molecules is currently of great interest for the design of new antitumor drugs. The interaction of a new metal complex with DNA was investigated through several techniques. Absorption spectroscopy and gel electrophoresis studies on the interaction of the Cu-complex of (2a-4mpyH)2 [Cu(pyzdc)2 (H2O)2].6 H2O with DNA have shown that this complex can bind to CT-DNA with binding constant 3.99 × 10(5) M(-1). The cyclic voltammetry (CV) responses of the metal complex in the presence of CT-DNA have shown that the metal complex can bind to CT-DNA through partial intercalation mode and this is consistent with molecular docking analysis, quenching process and thermal denaturation experiments. The cytotoxicity of this complex has been evaluated by MTT assay. The results of cell viability assay on DU145 cell line revealed that the metal complex had cytotoxic effects.

Developmental study of mercury effects on the fruit fly (Drosophila melanogaster).

Environmental pollution caused by heavy metals such as mercury is one of the most important human problems. It might have severe teratogenic effects on embryonic development. Some pharmacological and physiological aspects of fruit flies (Drosophila melanogaster) are similar to humans. So the stages of egg to adult fruit fly, as a developmental model, were employed in the study. Wild adult insects were maintained in glass dishes containing standard medium at 25 °C in complete darkness. Five pairs of 3-day old flies were then transferred to standard culture dishes containing different concentrations of mercury ion. They were removed after 8 hours. We considered the following: The rate of larvae becoming pupae and pupae to adults; the time required for the development; the hatching rate in the second generation without mercury in the culture; the morphometric changes during development in both length and width of the eggs through two generations; larvae, pupae and adult thorax length and width. The results showed that mercury in culture (20-100 mg/l) increase the duration of larvae (p<0.01) and pupae (p<0.01) development, the rate of larvae becoming pupae (p<0.001); pupae maturation (p<0.05), the hatching rate (p<0.01), the length (p<0.05) and width of larvae (p<0.01) and pupae (p<0.001) and the length in the adult thorax (p<0.01) decreased significantly. There was no effect upon the size of eggs. There were also no larvae hatching in concentrations of 200 mg/l of mercury. Negative effects of mercury as a heavy metal are possibly due to the interference of this metal in cellular signaling pathways, such as: Notch signaling and protein synthesis during the period of development. Since it bonds chemically with the sulfur hydride groups of proteins, it causes damage to the cell membrane and decreases the amount of RNA. This is the cause of failure of many enzyme mechanisms.