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An effective one-step electrochemical method was developed to synthesize three-dimensional N-doped porous graphene/magnetite nanoparticles hybrid onto Ni foam (Fe3O4@3D-NPG/NF electrode). In this method, 3D nitrogen-doped porous graphene layers are electrophoretically deposited onto Ni foam, accompanied by the simultaneous in situ electrochemical deposition (ECD) of magnetite particles onto 3D-NPG layers. For comparison, Fe3O4 particles and N-doped graphene were separately deposited onto Ni foam, and pristine Fe3O4/NF and 3D-NPG/NF electrodes were fabricated. The structure, composition and morphology of the fabricated electrode materials were systematically characterized by XRD, FT-IR, FE-SEM, Raman, TEM, BET, and TGA/DSC techniques. The formation mechanism of Fe3O4@3D-NPG hybrid through EPD/ECD was proposed and described in detail. The charge storage capabilities of the fabricated electrodes were analyzed as the supercapacitor electrode. The results GCD tests revealed that Fe3O4@3D-NPG electrode is able to deliver specific capacity value of 715 F g−1 at 2 A g−1 and cycle life of 94.3% after 5000 GCD cycles, where the pristine Fe3O4/NF electrode delivered only specific capacity of 219 F g−1 and 77.6% capacity retention. These findings implicated the positive synergistic effects between Fe3O4 and 3D-NPG in the hybrid electrode to exhibit higher supercapacitive performance. This simple strategy could find practical uses in the large-scale fabricating Fe3O4@3D-NPG electrode.

Purpose Recent studies show that the depletion of naturally occurring deuterium can result in tumor regression in both in vitro and in vivo models. In the present paper we report the inhibitory effect of different concentrations of DDW discretely and in combination with 5-FU on HT29 colon cancer cells and the possible mechanisms underlying these effects. Methods HT29 cells were treated with decreasing deuterium concentrations of DDW individually, 5-FU alone and both from 24 to 72 h. Cellular cytotoxicity of the cells were determined by the MTT proliferation assay. The morphology and structure of the cells were observed by scanning electron microscopy (SEM). Results We observed the changes in the cell cycle by flow cytometry and alterations in the cellular antioxidant enzymes by transmission chemiluminescence. Treatment with DDW especially in concentrations of 50 and 75ppm alone imposed inhibitory effects on the growth of the cell line and DDW augmented 5-FU inhibitory effects on HT29 cell lines. The cell cycle analysis revealed that DDW caused the cell cycle arrest in the G1/S transition, reduced the number of the cells in the S phase and significantly increased the population of cells in the G1 phase in HT29 cells. We observed that DDW induced the cellular antioxidant defense system. Under SEM examination, the control cells showed smooth surface and more extracellular matrix, while the DDW exposed group showed a rough surface and more microvillus on the cell surface. Conclusions We conclude that DDW is a promising new adjuvant anti-cancer agent with the potential for future clinical applications.

Synthesis of folic-acid mediated targeting nanocarrier is one of the most important and effective approaches in drug delivery. In this study, firstly, the surface of polyamidoamine G4 (PAMAM G4) dendrimers was modified with polyethylene glycol (PEG) to reduce dendrimer toxicity and was engineered with folic acid (FA) to increase nanocomplex targeting of the mice myoblast C2C12 cell lines. Both these synthetic phases were characterized via fourier transform infrared spectroscopy (FT-IR), Proton nuclear magnetic resonance (1H NMR), ultraviolet–visible spectrophotometry (UV–Vis), and differential scanning calorimetry (DSC) analysis. Secondly, 5-fluorouracil (5-FU), as a chemotherapeutic agent, was loaded in the nanocarrier to therapeutic goals and was characterized. In vitro drug release was also assessed and an excellent controlled release with good release efficiency was reported. The size, diameter, surface morphology, and surface charge of these synthesized nanocomplexes were ascertained with the scanning electron microscopy (SEM), dynamic light scattering (DLS) and zeta potential analyser, respectively, and PAMAM G4 positive charges eliminated. Finally, our nanocomplex was tested on the HT-29 human colorectal cancer and mice myoblast C2C12 normal cell lines and cell viability was evaluated with an 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. Good cancer cell inhibition and cell growth promotion effects of the nanocomplexes have been indicated in this research study.

Dendrimer-based targeted drug delivery, as an innovative polymeric drug-delivery system, is promising for cancer therapy. Folate receptors (FR) are overexpressed in many types of tumor cells, such as breast cell carcinomas, which allow folate-targeted delivery. Therefor polyethylene glycol (PEG) modified-PAMAM G4 dendrimers were functionalized with folic acid (FA), as targeting agent. Then, 5-FU (5-fluorouracil) and

Dendrimer-based targeted drug delivery, as an innovative polymeric drug-delivery system, is promising for cancer therapy. Folate receptors (FR) are overexpressed in many types of tumor cells, such as breast cell carcinomas, which allow folate-targeted delivery. Therefor polyethylene glycol (PEG) modified-PAMAM G4 dendrimers were functionalized with folic acid (FA), as targeting agent. Then, 5-FU (5-fluorouracil) and 99mTc (technetium-99 m) as therapeutic agents were respectively loaded and conjugated to previous nano-complex (PEG-PAMAM G4-FA-5FU-99mTc). The value of drug loading was calculated by TGA analysis (16.97%). Drug release profiles of PEG-PAMAM G4-FA-5FU-99mTc and PEG-PAMAM G4-FA-5FU were evaluated. The radiochemical purity of PEG-PAMAM G4-FA-5FU-99mTc and PEG-PAMAM G4-FA-99mTc was obtained at >95% with excellent in-vitro and in-vivo stabilities. PEG-PAMAM G4-FA-5FU-99mTc was synthesized and the stability studies were carried out by the ITLC methods in serum (86.67% and 83.75%) and PBS. Combinational therapy effects of 5-FU and 99mTc containing nano-complexes were evaluated on 4 T1 (mouse breast cancer) and MDA-MB-231 (human breast adenocarcinoma) cancer cell lines. Excellent uptake values were obtained for FA-decorated nano-complexes on 4 T1 and MDA-MB-231 cell lines. Subsequently, tumor inhibition effects of PEG-PAMAM G4-FA-5FU-99mTc and PEG-PAMAM G4-FA-5FU were evaluated using the breast tumor-bearing BALB/C mice.

B-cell lymphoma 2 (BCL2) family proteins play a critical role in tuning cell death processes. Almost in half of all human cancers, a dysregulation in BCL2 family gene expression has been shown which made it an impressive target for human gene therapy as a novel approach in cancers. In this study we will optimize lentiviral-mediated RNA interference (RNAi), recombinant lentiviruses accommodating anti-BCL2 micro adaptive short hairpin RNAs (shRNAs), to downregulate BCL2 in human embryonic kidney 293T (HEK293T) cells to produce stable cell lines. We tested 4 different Dharmaconâ„¢ GIPZâ„¢ shRNAmir lentiviral vectors targeting BCL2 in different positions and a pGIPZ non-silencing shRNAmir lentiviral vector (as a negative control). Lentivirus packaging was performed by the calcium phosphate precipitation method. HEK293T cells were transduced by each type of recombinant lentiviruses individually and selected by puromycin within 10 days. The relative mRNA level and protein expression were assayed by using real-time polymerase chain reaction (PCR) technic and western blotting, respectively. Lentivirus (LV) packaging was performed in high efficiency (transfection rate was > 90%). Recombinant viruses of 4 expression vector addition to a control vector were produced then transduced to HEK293T cells successfully. All the 4 cell groups showed a significant down regulation of BCL2 gene (~90-95%) at mRNA level compared to the control group (p 0.05). We showed that the lentivirus-mediated RNAi technique is an efficient method to establish HEK293 cell lines with stable down-regulation of BCL2 gene.

Overexpression of folic acid receptor in various human tumors cells makes it as good candidate for targeting delivery of chemotherapeutic and radiopharmaceutical agents. In this research, FA used for functionalization of PEG modified PAMAM G4 dendrimer as a smart delivery of 5-FU and 99m Tc for the breast carcinoma in order to chemotherapeutic and imaging goals. One aim of this research was assess the FA-mediated cell viability assay of PEG-PAMAM G4-FA-5FU- 99m Tc and in vitro uptake of PEG-PAMAM G4-FA- 99m Tc as the novel nano-complex determined on C2Cl2 (normal cell) and MCF-7 (breast cancer cell) cell lines. Other main goals were studied. Morover, an investigation in to in vivo imaging and biodistribution was carried out via a novel radio tracer by which tumor accumulation and site were obviously detected. The targeted tumor images taken by tail intravenous injection demonstrated that nano-complex can be smartly used in imaging study of the clinical practices. Also, the biodistribution of this nano-complex was investigated and the organ predestination of 99m Tc labeled nano-complex (%ID/g) was ascertained.

Recent studies have shown that the depletion of naturally occurring deuterium can result in tumor regression. The aim of the present study was to show the growth inhibitory effects of DDW discretely and in combination with 5-FU on MCF-7 breast cancer cells and to determine possible mechanisms underlying these changes. MCF7 cells were grown in RPMI medium with decreasing deuterium concentrations of DDW individually, 5-FU alone and both for 24, 48, and 72 h. Cell viability was determined with the MTT assay. The cell cycle and antioxidant enzymes status were measured using flow cytometry and quantitative luminescence methods, respectively. Our results showed that treatment with DDW especially in 30-100 ppm concentrations imposed the highest cell growth inhibitory effect. The cell cycle analysis revealed that DDW caused the cell cycle arrest in the

In this work, binder-free composite of Co(OH)2 nanosheets/oxygen-functionalized g-C3N4 (f-g-C3N4) was co-embedded onto nickel foam through a simple one-step deposition process. In this process, f-g-C3N4 and Co(OH)2 are electrochemically co-embedded into Ni-Foam support using a simple electrodeposition procedure in a two-electrode system from an aqueous electrolyte containing both f-g-C3N4 and cobalt nitrate salt. FE-SEM and TEM observations confirmed that Co(OH)2 nanosheets are hierarchically formed on the surface of porous f-g-C3N4 electrophoretically deposited onto the surface of nickel foam making unique three-dimensional porous networks. The specific capacitances of Co(OH)2@f-g-C3N4/Ni-foam hybrid electrode were measured to be 1663 F g–1 and 1064 F g–1 at 1 A g–1 and 30 A g–1, respectively, which are higher than those of pristine Co(OH)2/Ni-foam electrode (i.e. 1267 F g–1 and 462 F g–1 at 1 A g–1 and 30 A g–1, respectively). Furthermore, nanocomposite electrode exhibited high capacitance retentions i.e. 64.1% retention with increasing the applied load from 1 A g–1 to 30 A g–1, also 93.4% and 88.6% capacity stabilities after 5000 cycling at 2 A g–1 and 6 A g–1, where pristine Co(OH)2/Ni foam showed only 36.5% retention at high-rate discharging, and capacity stabilities of 84.6% and 69.1% at 2 A g–1 and 6 A g–1, respectively. These improvements were ascribed to the synergetic effects between Co(OH)2 nanosheets and oxygen-functionalized g-C3N4, which supported the fabricated nanocomposite as a new high-performance electrode material for supercapacitors.

PURPOSE Serum and intracellular instability limits the therapeutic applications of short interfering RNA (siRNA) as a radiopharmaceutical. Chemical modifications like phosphorothioate (PS) substitution and 2'-O-methoxy (2'-O-Me) modifications could eliminate such limitations. In this study, the effects of PS and 2'-O-Me modifications at the backbone of siRNA on serum stability and RNA interference activity were investigated. MATERIALS AND METHODS Fully PS and 2'-O-Me-modified type 1 insulin-like growth factor receptor (IGF-1R) siRNA was radiolabeled with lutetium-177 ((177)Lu) through p-SCN-Bn-DTPA as a chelator. After purification with Vivaspin and PD-10 columns, the radiolabs were examined for stability in serum by instant thin-layer chromatography and polyacrylamide gel electrophoresis. The level of IGF-1R in response to the modified and labeled IGF-1R siRNA was examined using RT-PCR and ELISA assay in colon cancer cells. The effects of such siRNA on the prevention of proliferation of colon cancer cells and its apoptosis were investigated using MTT assay and Annexin-V/propidium iodide double staining, respectively. Cellular accumulation quantities of the labeled and modified IGF-1R siRNA were determined using a γ-counter by taking advantage of (177)Lu as a γ-emitter. RESULTS Both the modified (177)Lu-siRNA complex and the modified nonlabeled siRNA showed significant stability in serum. The levels of IGF-1R mRNA and protein significantly decreased with both the labeled and nonlabeled IGF-1R siRNAs, but no such reduction in IGF-1R was observed with luciferase siRNA (P

Cerium-141 [T 1/2 = 32.5 days, β 1 = 580.4 keV (30 %), β 2 = 435.0 keV (70 %), γ = 145.4 keV (48 %)], has been introduced as a radionuclide for therapy while can be used in diagnosis as well. In this study, nuclear model calculation on 141Ce production was investigated via the 140Ce(d,p)141Ce, 142Ce(d,dn)141Ce, 141Pr(n,p)141Ce, and 140Ce(n,γ)141Ce nuclear reactions. 140Ce was irradiated by thermal neutron at the Tehran Research Reactor according to the 140Ce(n,γ)141Ce reaction. In addition, the obtained activity of the produced 141Ce was compared with the theoretical calculations. The results showed that the end of bombardment activities of 141Ce is 631.64 MBq theoretically and 611.60 MBq experimentally for ceria powder. The activities of similar samples of ceria (CeO2) powder in two forms of nano-particles (nanoceria) and bulk were compared after bombardment by thermal neutrons. The results showed that activities of nanoceria were less than the bulk form of ceria.

Blood and the system that forms it, known as the hematopoietic system, consist of many cell types with specialized functions. Redblood cells (erythrocytes) carry oxygen to the tissues. Platelets (derived frommegakaryocytes) help prevent bleeding. Granulocytes(neutrophils, basophils and eosinophils) and macrophages (collectively known as myeloid cells) fight infections from bacteria, fungi,and other parasites such as nematodes (ubiquitous small worms). Some of these cells are also involved in tissue and bone remodelingand removal of dead cells. B-lymphocytes produce antibodies, while T-lymphocytes can directly kill or isolate by inflammation cellsrecognized as foreign to the body, including many virus-infected cells and cancer cells. Many blood cells are short-lived and need to bereplenished continuously; the average human requires approximately one hundred billion new hematopoietic cells each day. The continuedproduction of these cells depends directly on the presence of Hematopoietic Stem Cells (HSCs), the ultimate, and only, sourceof all these. Peripheral blood stem cells (PBSC) are rapidly becoming the primary rescue modality for autologous transplantation andare now actively being investigated in the allogeneic transplant setting. Many investigators and clinical researchers believe that PBSCare likely to replace bone marrow stem cells entirely, for use in clinical transplantation in the not too distant. Hematopoietic stem cells(HSCs) are the blood cells that give rise to all the other blood cells. They give rise to the myeloid (monocytes and macrophages, neutrophils,basophils, eosinophils, erythrocytes, megakaryocytes/platelets, dendritic cells), and lymphoid lineages (T-cells, B-cells, NK-cells).The definition of hematopoietic stem cells has changed in the last two decades. The hematopoietic tissue contains cells with long-termand short-term regeneration capacities and committed multipotent, oligopotent, and unipotent progenitors.

Summary In this study, bevacizumab was successively labeled with 111In-InCl3 after conjugation with DOTA-NHS-ester followed by molecular filtration and determination of the average number ofDOTAconjugated per mAb (6 : 1) by spectrophotometric method. Radiochemical purity (> 97%, measured by ITLC and HPLC), integrity of protein after radiolabeling (gel electrophoresis) and stability of 111In-DOTA-Bevacizumab (in final formulation, human serum, liver/kidney homogenates) were determined in 24-72 h as well as biodistribution studies in wild-type rats and human colon cancer (SW-480) bearing mice. The accumulation of the radiolabeled antibody was consistent with the former reported Bevacizumab conjugates. Significant tumor uptake (8%) was observed at 72 h p.i. Tumor/muscle uptake ratios were 2.6 (24 h), 9.74 (48 h) and 25 (72 h). 111In- DOTA-Bevacizumab was prepared as a SPECT molecular imaging agent for diagnosis and follow-up of vascular endothelial growth factor A (VEGF-A) expression in oncology.

Purpose The radiolabeling of targeting biomolecules with gamma emitter radionuclides for tracing and beta emitters for therapy involves the conjugation of such biomolecules to the chelating agents to form complexes with the radionuclide of interest. In this study, radioconjugate of IGF-1R siRNA with lutetium-177 ( 177 Lu) was produced, and the anti-proliferation and apoptosis effects elicited by this 177 Lu-siRNA complex in the SW480 colon cancer cells were evaluated. Methods IGF-1R and Luciferase siRNAs were conjugated with p-SCN-Bn-DTPA, and then radiolabeled with 177 Lu. The effects of labeled and non-labeled IGF-1R siRNAs on IGF-1R expression were assessed with RT-PCR analysis and ELISA assay. IGF-1R siRNAs induced cell death and apoptosis were investigated using MTT assay and Annexin-V/propidium iodide (PI) double staining, respectively. Results Combined purification using Vivaspin and PD-10 columns resulted in a radiochemical purity of 97.32%±1.97%. Knockdown effect of the labeled IGF-1R siRNA was not significantly different from the non-labeled duplex of the same sequence (P 177 Lu-IGF-1R siRNA in comparison with either 177 Lu or IGF-1R siRNA (P Conclusion Radioconjugate of IGF-1R siRNA, p-SCN-Bn-DTPA and 177 Lu was successfully produced and characterized as radiopharmaceutical. The present study demonstrates the involvement of 177 Lu-labeled IGF-1R siRNA in the inhibition of cell growth and induction of apoptosis in colon cancer cells.

It is usually believed that γ-ray interaction with biomolecules is intermediately performed by reactive oxygen species (ROS) produced from radiolysis of water. Hemoglobin (Hb) as one of the most abundant biomolecule in blood and well-studied endogenously affected by ROS, was a good candidate for study. Adult human Hb was extracted and irradiated using four distinct 20, 60, 90 and 170 Gy doses from Co-60 γ-ray source. UV–vis, fluorescence and FT-IR spectroscopies were used to study the whole conformational changes and partial degradation of heme. Hb species calculated using Benesch equations indicated that the concentration of oxy-Hb was decreased from 9.97 μM to 6.56 μM, while the total metastable met and deoxy-Hb concentration were just increased 2.39 μM and about 8.4% of total heme was diminished. Heme degradation was studied using fluorescence spectra at two 321 and 460 nm excitation wavelengths as fully and partially degradation of heme respectively. Inverse behavior of these two fluorescence spectra suggested a new mechanism of heme degradation in which γ-ray preferably absorbed by heme without any intermediary effects of water. It was confirmed by FT-IR spectra at 900–1000 cm −1 where the Fe N and N H of porphyrin indicate their own stretching vibrational bands. Thermal stability justified that the gamma radiation induced the conformational changes of Hb which is appeared during thermal unfolding. First derivative of thermal spectra indicated that the Tm of 170 Gy dose irradiated sample is 2 °C lowered and total concentration of Hb was decreased 14%.

Insulin-like growth factors are among the peptide mitogens that regulate cell proliferation and differentiation as well as mediator of antiapoptotic signals. The imbalance between the expression and activities of these molecules may lead to malignancy in cells. Evidences have suggested the insulin-like growth factor 1 receptor (IGF-1R) signaling pathway as a therapeutic target in the management and treatment of cancer. In this present study, we have generated silencing stable clones of HEK cells using six different pGIPZ (lentiviral vector) shRNAs targeted to human IGF-1R gene and a pGIPZ non-silencing shRNAmir lentiviral vector (as negative control). The recombinant lentiviral vectors were separately transduced into human embryonic kidney 293 T (HEK293T) cell lines. The knockdown of IGF-1R was confirmed by reverse transcription polymerase chain reaction (RT-PCR) and the relative IGF-1R mRNA levels were expressed as a ratio of IGF-1R to β-actin by REST software. The results showed significant reduction in the expression of IGF-1R mRNAs in cells transduced with all six pGIPZ-IGF-1R recombinant lentivirals compared to non-silencing negative control. No significant difference was observed among the six cassettes. Results indicated that recombinant lentiviral vectors provided an efficient and stable knockdown of IGF-1R providing useful tool for IGF-1R pathway studies.

Overexpression of folic acid receptor in various human tumors cells makes it as good candidate for targeting delivery of chemotherapeutic and radiopharmaceutical agents. In this research, FA used for functionalization of PEG modified PAMAM G4 dendrimer as a smart delivery of 5-FU and

Yttrium-90 is a useful radionuclide for radioimmunotherapy (RIT) and the anti-epidermal growth factor receptor (anti-EGFR) antibody cetuximab is clinicsally approved for the treatment of EGFR-expressing metastatic colorectal cancer and advanced head and neck cancer. Thus in this work radiolabeling of monoclonal anti-EGFR with 90Y for radioimmunotherapy (RIT) is targetted. Cetuximab was successively labeled with [90Y] chloride (74 MBq) 2 mCi after conjugation with macrocyclics bifunctional chelating agent, 1,4,7,10-tetraazacyclododecane-N,N′,N″,N″′-tetraacetic acid mono-(N-hydroxysuccinimidyl) ester (DOTA-NHS), purified and concentrated by centrifugation using an Amicon Ultra-15 filter (Millipore, MWCo, 30000). 90Y chloride was obtained by 90Sr/90Y generator. Radiolabeling was completed in 2 h by the addition of DOTA-cetuximab conjugate at 42 °C. The stability of radiolabeled was studied in human serum. Biodistribution studies in normal rats were carried out to determine the radioimmunoconjugate distribution up to 96 h. Radiochemical purity of 92 % (using ITLC) was obtained for final radioimmunoconjugate (Specific activity = 0.55 GBq/mg). Stability of radiolabeled protein in presence of human serum was tested at 37 °C for up to 24 h. Biodistribution studies demonstrated the highest ID/g % in the blood (2.62 ± 0.005 at 24 h) and the liver (2.19 ± 0.001). This study demonstrated that 90Y-DOTA-cetuximab is a potential compound for the treatment of EGFR-expressing cancers.

Gold nanorods (GNRs) can be used in various biomedical applications; however, very little is known about their in vivo tissue distribution by radiolabeling. Here, we have developed a rapid and simple method with high yield and without disturbing their optical properties for radiolabeling of gold rods with iodine-131 in order to track in vivo tissue uptake of GNRs after systemic administration by biodistribution analysis and γ-imaging. Following intravenous injection into rat, PEGylated GNRs have much longer blood circulation times.

Breast cancer radioimmunoscintigraphy targeting HER2/neu expression is a growing field of work in nuclear medicine research. Trastuzumab is a monoclonal antibody that binds with high affinity to HER2/neu, which is over expressed on breast and other tumors. Developing new tracers for the detection of this cancer is of great interest. In this study, trastuzumab was successively labeled with [64Cu]CuCl2 after conjugation with DOTA-NHS-ester. The conjugate was purified by molecular filtration, the average number of DOTA conjugated per mAb was calculated and total concentration was determined by spectrophotometric method. DOTA–trastuzumab was labeled with 64Cu produced by 68Zn(p,αn)64Cu nuclear reaction (30 MeV protons at 180 μA). Radiochemical purity, integrity of protein after radiolabeling and immunoreactivity of radiolabeled mAb trastuzumab with HER2/neu antigen and SkBr3 cell line were performed by RIA. In vitro stability of radiolabeled mAb in human serum was determined by thin layer chromatography. In vitro internalization studies were performed with the SkBr3 cell line and the tissue biodistribution of the 64Cu–DOTA–trastuzumab was evaluated in wild-type rat (90 ± 5.5 μCi, 2, 6, 12, 24 h p.i.). The radioimmunoconjugate was prepared with a radiochemical purity of higher than 96 ± 0.5 % (ITLC) and specific activity as high as 5.3 μCi/μg. The average number of chelators per antibody for the conjugate used in this study was 5.8/1. The sample was showed to have similar patterns of migration in the gel electrophoresis. The 64Cu–DOTA–trastuzumab showed high immunoreactivity towards HER2/neu antigen and SkBr3 cell line. In vitro stability of the labeled product was found to be more than 94 % in PBS and 82 ± 0.5 % in human serum over 48 h. In vitro internalization studies of the 64Cu–DOTA–trastuzumab showed that up to 11.5 % of the radioimmunoconjugate internalized after 10 h. The accumulation of the radiolabeled mAb in liver, skin, intestine, lung, spleen, kidney and other tissues demonstrates a similar pattern to the other radiolabeled anti-HER2 immunoconjugates. 64Cu–DOTA–trastuzumab is a potential compound for molecular imaging of PET for diagnosis and treatment studies and follow-up of HER2 expression in oncology.

Antiangiogenic monoclonal antibodies in combination with therapeutic radionuclides are potential targeted therapy agents in cancer. In this study, bevacizumab was successively labeled with [166Ho]HoCl3 after conjugation with DOTA-NHS-ester with a radiochemical purity of higher than 95% (RTLC). The conjugates were purified by molecular filtration, the average number of DOTA conjugated per mAb was calculated and total concentration was determined by spectrophotometric method and the average chelate to antibody ratio (c/a) for the conjugate used in this study was 5.8:1 and protein integrity experiments (SDS-PAGE). The biodistribution studies in wild-type rats demonstrate a similar pattern to the other radiolabeled anti-vascular endothelial growth factor A (VEGF-A) immunoconjugates. 166Ho-DOTA-bevacizumab is a potential compound for therapy/imaging of VEGF-A expression in oncology.

Developing monoclonal antibodies labeled with beta-emitters has led to the introduction of important agents in radioimmunotherapy. In this work, Sc-46 chloride was obtained by thermal neutron activation flux of natural metallic scandium sample followed by dissolution in acidic media (radionuclidic purity via beta and gamma ray spectroscopy, 99.9% ; radiochemical purity via ITLC, >99%) and used in radiolabeling of rituximab after conjugation with DOTA-NHS-ester. The specific activity was however not high. The conjugates were purified by molecular filtration and used in the radiolabeling. The radiochemical purity (ITLC), stability studies (ITLC and size exclusion chromatography), determination of average number of DOTA conjugated per mAb (chelate: antibody ratio, 5.8:1) and gel electrophoresis of [ 46Sc]Sc-DOTA-anti-CD20 were determined followed by biodistribution studies for 46Sc and [ 46Sc]Sc-DOTA-anti-CD20 i n wild type rats up to 72 h. The binding of the radiolabeled antibody was showed to be 60% on Raji cells. The final compound was stable in presence of PBS at 37 ºC and room temperature. The accumulation of the radiolabeled antibody in liver, spleen, kidney, heart and other tissues demonstrates a pattern similar to the other radiolabeled anti-CD20 immunoconjugates. The present study shows the possibility of antibody labeling for future use in radioimmunotherapy by 47Sc.

Colon cancer is the second leading cause of cancer death worldwide. Elevated expression of insulin-like growth factor-I receptor (IGF-IR) is a frequent genetic abnormality seen in this malignancy. For a better understanding of its role in maintaining the malignant phenotype, we used RNA interference (RNAi) directed against IGF-IR in our study. The aim of this study was to examine the anti-proliferation and chemosensitization effects elicited by a decrease in the transcription and protein levels of IGF-IR by RNAi in SW480 colon cancer cells. A plasmid-based polymerase III promoter system was used to deliver and express short interfering RNA (siRNA) targeting IGF-IR to reduce its expression in SW480 cells. Western blot analysis was used to measure the protein level of IGF-IR. We assessed the effects of IGF-IR silencing on cancer cell growth by a cell growth curve. The effect of the 5-fluorouracil (5-FU)-induced cell death by knockdown of IGF-IR was also investigated by methyl thiazolyl tetrazolium assay. Transfection of siRNA targeting IGF-IR was shown to reduce IGF-IR messenger RNA levels by 95%. Western blotting detected a similar inhibition of IGF-IR protein levels in those cells. The cells transfected with PKD-short hairpin RNA-IGF-IR-V2 significantly decreased cell growth and rendered cells more sensitive to chemotherapy. The highest proliferation inhibitory and chemosensitization ratios were 53 ± 2% and 1.78, respectively. This study indicates that downregulation of IGF-IR results in significant inhibition of tumor growth in vitro. It also provides a promising strategy to chemotherapy efficacy in human tumors and forming a basis for future in vivo trials.

Background and Aims Colon cancer is the second leading cause of death due to cancer worldwide. Elevated expression of IGF-IR is a frequent genetic abnormality seen in this malignancy. The aim of the study was to examine the anti-growth effects elicited by a decrease in the protein level of IGF-IR by RNA interference (RNAi) in SW480 cells. Methods A plasmid-based polymerase III promoter system was used to deliver and express short interfering RNA (siRNA) targeting IGF-IR to reduce its expression in SW480 cells. The expression of IGF-1R protein was detected by Western blot. We assessed the effects of IGF-IR silencing on cancer cell growth by a growth curve. Results We prepared a type of IGF-IR short hairpin RNA (shRNA) expression vector that could efficiently inhibit the expression of IGF-IR in SW480 cells. At 48 h after transfection, the expression inhibition rate was 92 ± 2% at mRNA level detected by RT-PCR analysis. Western blotting detected a similar inhibition of IGF-IR protein levels in cells transfected with pkD-shRNA-IGF-IR-V2. Downregulation of IGF-IR resulted in significant inhibition of cancer cell growth in vitro . The cell growth inhibition rates at 24, 48, and 72 h after pkD-shRNA-IGF-IR-V2 transfection were 32.06, 47.61, and 35.36%, respectively. Conclusions Our data show that decreasing the IGF-IR protein level in SW480 cells by RNAi could significantly inhibit tumor growth in vitro , implying the therapeutic potential of RNAi on the treatment of colon cancer by targeting overexpression oncogenes such as IGF-IR. IGF-IR may be a potential therapeutic target for human colon cancer.

Background Protein tyrosine phosphatase 1B (PTP1B), encoded by the PTPN1 gene, efficiently dephosphorylates the insulin receptor, and attenuates insulin signaling. Recently, a 1484insG variant of the PTPN1 gene has been associated with an increased risk of metabolic syndrome in an Italian population that has not been confirmed in the subsequent studies. The purpose of this study was to investigate the association of 1484insG polymorphism of the PTPN1 with obesity, insulin resistance, type 2 diabetes and other cardiovascular-related traits in an Iranian population. Methods The genotypes of 1484insG variant were determined by PCR-RFLP method in 696 unrelated subjects including 412 subjects with normal glucose tolerance and normal fasting glucose and 284 type 2 diabetic patients. Results The allelic frequency of 1484insG polymorphism among type 2 diabetic patients and non-diabetic subjects was 4.9 and 4.1%, respectively ( p = 0.475). In type 2 diabetic patients, among quantitative traits, no significant difference in anthropometric and biochemical parameters was seen between the wild-type and heterozygous 1484insG genotypes in male and female groups and in non-diabetic subjects, male carriers of 1484insG allele had significantly higher fasting insulin ( p = 0.003), cholesterol ( p = 0.012), LDL-C ( p = 0.037), apo B ( p = 0.015), and HOMA-IR ( p = 0.011) levels compared to the individuals carrying the wild-type genotype. Among non-diabetic female individuals, only body mass index was significantly higher in 1484insG subjects compared to the wild-type individuals ( p = 0.007). Conclusions Our results from a sample of Iranian type 2 diabetes cases and controls provide evidence that the 1484insG genotype of the PTPN1 gene may be associated with insulin resistance and other cardiovascular risk factors in non-diabetic male subjects.

Microsphere and particle technology with selective transport of radiation represents a new generation of therapeutics. Poly-L-lactic acid (PLLA) microspheres loaded with holmium-166 acetylacetonate ((166)Ho-PLLA-MS) are novel microdevices. In this research, (165)HoAcAc-PLLA microparticles were prepared by the solvent evaporation technique. Microspheres were irradiated at Tehran Research Reactor. The diameter and surface morphologies were characterized by particle sizer and scanning electron microscopy before and after irradiation. The complex stability, radiochemical purity, and in vivo biodistribiotion were checked in the final solution up to 3 days. In this study, (166)Ho-PLLA spherical particles with a smooth surface and diameter of 20-40 µm were obtained, which were stable in vitro and in vivo studies. Neutron irradiation did not damage the particles. The ease with which the PLLA spheres could be made in the optimal size range for later irradiation and their ability to retain the (166)Ho provided good evidence for their potential use in radioembolization.

Dental pulp is the soft live tissue inside a tooth. Dental pulp contains stem cells, known as Dental Pulp Stem Cells. The finest Dental Pulp Stem Cells are found in a baby teeth or milk teeth. The stem cells from the milk teeth are ‘mesenchymal’ type of cells. cells that have the ability to generate a wide variety of cell types like chondrocytes, osteoblasts and adipocytes. To isolate high-quality human dental pulp stem cells from accessible resources is an important goal for stem-cell research. In this study we found that exfoliated human deciduous tooth contains multipotent stem cells, they identified to be a population of highly proliferative, clonogenic cells. Hematopoietic stem cells (HSCs) are the blood cells that give rise to all the other blood cells. HSCs are found in the bone marrow of adults, with large quantities in the pelvis, femur, and they are also found in umbilical cord blood and, in small numbers, in blood. In this study we differentiated stem cells that isolated from human dental pulp and peripheral blood to cardiac cells. The isolation process for dental pulp stem cells would take place in 2 different methods: The enzymatic digestion method and, the second method is a subsidiary one. For isolating the stem cells from peripheral blood we use ficoll method. Flow cytometery analyzes showed that dental pulp stem cells are mesanchymal and peripheral blood cells are hematopoietic stem cells. Bothe group of stem cells can differentiation to cardiac cells. Dental pulp stem cells are mesanchymal cells and peripheral blood cells are hematopoietic cells. For differentiation the cells to cardiomyocite we used 5-azacitidin and exitocin. After differentiation stem cells to cardiac cells, dental pulp stem cells and peripheral blood stem cells morphology was changed, before differentiation to cardiac cells, both group were elongated and after differentiated to cardiomyocit they have central typical nuclear and were stratified ,they expanded cytoplasmic papillae .

Gold nanorods (GNRs) can be used in various biomedical applications; however, very little is known about their in vivo tissue distribution by radiolabeling. Here, we have developed a rapid and simple method with high yield and without disturbing their optical properties for radiolabeling of gold rods with iodine-131 in order to track in vivo tissue uptake of GNRs after systemic administration by biodistribution analysis and γ-imaging. Following intravenous injection into rat, PEGylated GNRs have much longer blood circulation times.

Insulin like growth factor receptor 1 (IGF-1R) is well-documented to play a key role in radiation response and tumor radiosensitivity, thus offering an attractive clinic drug target to enhance tumor sensitivity to anti-cancer radiotherapy.Human colon carcinoma SW480 cells were transfected with the specific small interference RNA (siRNA) expression vector (pkD-shRNA-IGF-1R-V2) designed to target IGF-1R mRNA. The expression of IGF-1R mRNA and its protein among the transfected and untransfected cells were detected by semi-quantitative RT-PCR and ELISA assay. The changes in cell radiosensitivity were examined by MTT assay.Transfection of mammalian expression vector pkD containing IGF-1R siRNA was shown to reduce IGF-1R mRNA levels by up to 95%. ELISA assay detected a similar inhibition of IGF-1R protein levels in cells transfected with IGF-1R siRNA. SW480 cells transfected with the expression vector for siRNA significantly rendered cells more sensitive to radiation and the highest radiation enhancement ratio was 2.02 +/- 0.08.These data provide the first evidence that specific siRNA fragment (pkD-shRNA-IGF-1R-V2) targeting human IGF-1R mRNA is able to enhance colon cancer radiosensitivity. Also results indicated that, combining IGF-1R siRNA and radiation significantly enhances antitumor efficacy compared with either modality alone.

Background On the basis of results of our previous investigations on 90Y-DTPA-rituximab and in order to fulfil national demands to radioimmunoconjugates for radioscintigraphy and radioimmunotherapy of Non-Hodgkin’s Lymphoma (NHL), preparation and radiolabeling of a lyophilized formulation (kit) of DOTA-rituximab with 111In and 90Y was investigated. Methods 111In and 90Y with high radiochemical and radionuclide purity were prepared by 112Cd (p,2n)111In nuclear reaction and a locally developed 90Sr/90Y generator, respectively. DOTA-rituximab immunoconjugates were prepared by the reaction of solutions of p-SCN-Bz-DOTA and rituximab in carbonate buffer (pH = 9.5) and the number of DOTA per molecule of conjugates were determined by transchelation reaction between DOTA and arsenaso yttrium(III) complex. DOTA-rituximab immunoconjugates were labeled with 111In and 90Y and radioimmunoconjugates were checked for radiochemical purity by chromatography methods and for immunoreactivity by cell-binding assay using Raji cell line. The stability of radiolabeled conjugate with the approximate number of 7 DOTA molecules per one rituximab molecule which was prepared in moderate yield and showed moderate immunoreactivity, compared to two other prepared radioimmunoconjugates, was determined at different time intervals and against EDTA and human serum by chromatography methods and reducing SDS-polyacrylamide gel electrophoresis, respectively. The biodistribution of the selected radioimmunoconjugate in rats was determined by measurement of the radioactivity of different organs after sacrificing the animals by ether asphyxiation. Results The radioimmunoconjugate with approximate DOTA/rituximab molar ratio of 7 showed stability after 24 h at room temperature, after 96 h at 4°C, as the lyophilized formulation after six months storage and against EDTA and human serum. This radioimmunoconjugate had a biodistribution profile similar to that of 90Y-ibritumomab, which is approved by FDA for radioimmunotherapy of NHL, and showed low brain and lung uptakes and low yttrium deposition into bone. Conclusion Findings of this study suggest that further investigations may result in a lyophilized (kit) formulation of DOTA-rituximab which could be easily radiolabeled with 90Y and 111In in order to be used for radioimmunotherapy and radioscintigraphy of B-cell lymphoma in Iran.