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Introduction and Aim: Breast cancer (BC) is one of the top three common cancers in women, responsible for nearly one-third of all new cancer diagnoses. Angiogenesis plays a crucial role in BC progression. In this study, we aimed to measure the serum concentrations of eight angiogenic factors in BC patients and healthy controls and to assess their correlation with clinicopathological variables., Methods: In a case-control study, 62 pathologically confirmed BC patients as well as 54 age-matched controls were recruited. A bead-based immunoassay was used to measure serum levels of VEGF-A, ANG-2, PDGF-AA, PDGF-BB, EGF, TGF-α, HGF, and bFGF., Results: We observed a significant elevation in serum levels of VEGF-A, EGF, and PDGF-AA in BC patients compared with the controls (P < 0.05). Patients with grade III had higher ANG-2 levels compared with those with grades I (P = 0.007) and II of the disease (P = 0.003). In addition, estrogen-positive and progesterone-positive BC patients had higher levels of TGF-α (P < 0.05)., Conclusion: The significant elevation of VEGF-A, EGF, and PDGF-AA serum levels in BC patients suggests these cytokines might have diagnostic value as potential biomarkers in BC. Further large-scale studies are needed to generalize these results to all BC patients.

The complex [(PhCH 2 NC)AuCl], 1 , was prepared by the reaction of [(Me 2 S)AuCl], A , with an equimolar amount of benzyl isocyanide (PhCH 2 NC) ligand. Through a salt metathesis reaction, the chloride ligand in 1 was replaced by potassium benzothiazole-2-thiolate (Kbt) and potassium benzoimidazole-2-thiolate (Kbi) to afford complexes (PhCH 2 NC)Au(κ 1 -S-bt)], 2a and (PhCH 2 NC)Au(κ 1 -S-bi)], 2b , respectively, which were characterized by NMR spectroscopy. The cytotoxic activities of 2a and 2b were evaluated against three human cancer cell lines, including A549 (lung), SKOV3 (ovary), and MCF-7 (breast). Our results indicated that 2a exhibited comparable cytotoxicity on investigated cell lines with cisplatin. It showed a good anti-proliferative activity with IC 50 of 19.46, 11.76 and 13.27 μM against A549, SKOV3 and MCF-7 cell lines, respectively. The effects of these complexes on the proliferation of the non-tumorigenic epithelial breast cell line (MCF-10A) showed their good selectivity between the tumorigenic and non-tumorigenic cell lines. Molecular docking simulation studies were also conducted to determine the specific binding site and binding mode of the synthesized gold complexes to DNA and thioredoxinreductase (TrxR) as their proposed targets.

Antibody-conjugated magnetic nanoparticles (Ab-MNPs) have received considerable attention in bioseparation and clinical diagnostics assays due to their unique ability to detect and isolate a variety of biomolecules and cells. Because antibodies can be expensive, a key challenge for bioconjugation is to determine the optimal amount of antibodies with reasonable antigen-capturing activity. We designed an approach to determine the minimum amounts of antibodies for efficient coating. Different quantities of Herceptin (anti-human epidermal growth factor receptor 2: HER2) antibody were applied and immobilized on the surface of MNPs. Antibody binding was then checked by using an anti-human antibody conjugated with fluorochrome and flow cytometry. When the ratio of MNPs to antibodies increased from 0.79 to 795.45, mean fluorescence intensity (MFI) of conjugated MNPs decreased markedly from 185.56 to 20.07, indicating lower surface antibody coverage. We then investigated the relation between antibody content and isolation efficiency. Three Ab-MNP samples with different MFI were used to isolate SK-BR-3, a HER2-positive breast cancer cell line, from mixtures of whole blood or mononuclear cells. After isolation in a magnetic field, separation efficiency was evaluated by fluorescence microscopy and flow cytometry-based techniques. Our results collectively showed that the amount of anti-HER2 antibodies for conjugation with MNPs could be decreased by as much as one-fifteenth without compromising isolation efficiency, which in turn can reduce the cost of immunoassay biosensors., (© 2020 Published by Elsevier Ltd.)

Background: Human immunological memory is a hallmark of the adaptive immune system and plays an important role in the development of effective immune responses against tumors. In the present study, we aimed to determine the frequencies of CD8 + memory T cell subsets including T stem cell memory (TSCM) in tumor-draining lymph nodes of patients with breast cancer (BC)., Methods: Mononuclear cells were obtained from axillary lymph nodes of 52 untreated patients with BC and stained for CD8, CCR7, CD45RO, CD95 markers to detect different subtypes of memory cells in the CD8 + lymphocyte population. Data were acquired on four-color flow cytometer and analyzed with CellQuest Pro software., Results: We observed that 47.65 ± 2.66% of CD8 + lymphocytes expressed the CD45RO, a marker for memory T cells. Statistical analysis showed that the total frequency of central memory T cells (TCM) and their subset with low CD45RO expression was significantly higher in tumor-involved nodes compared to tumor-free ones (P = 0.024 and P = 0.017, respectively). The level of CD95 expression (based on mean fluorescence intensity) on the surface of TCM, their CD45RO hi and CD45RO low subsets, and TSCM was higher in patients with stage II compared to those in stage I (P < 0.05). In addition, the percentage of naive CD8 + T cells was significantly lower in tumor-involved lymph nodes compared to tumor-free ones (P = 0.025)., Conclusions: Our data collectively indicate no significant differences in the frequencies of CD8 + lymphocytes or their memory subsets in tumor-draining lymph nodes of patients with BC. However, the frequency of CD45 low TCM was higher in tumor-involved nodes. Along with a decrease in the frequency of naive T cells, the higher frequency of CD45 low TCM suggests that despite the immune reaction to provide a pool of effective memory cells, it is blocked in early-stage of memory cells' differentiation (CD45RO low ), probably by tumor-derived suppressive factors. Identifying the molecular and cellular mechanisms behind this suppression can provide invaluable tools for adoptive T cell therapies in cancer.

Although offshore freshened groundwater (OFG) systems have been documented in numerous continental margins worldwide, their geometry, controls and emplacement dynamics remain poorly constrained. Here we integrate controlled-source electromagnetic, seismic reflection and borehole data with hydrological modelling to quantitatively characterise a previously unknown OFG system near Canterbury, New Zealand. The OFG system consists of one main, and two smaller, low salinity groundwater bodies. The main body extends up to 60 km from the coast and a seawater depth of 110 m. We attribute along-shelf variability in salinity to permeability heterogeneity due to permeable conduits and normal faults, and to recharge from rivers during sea level lowstands. A meteoric origin of the OFG and active groundwater migration from onshore are inferred. However, modelling results suggest that the majority of the OFG was emplaced via topographically-driven flow during sea level lowstands in the last 300 ka. Global volumetric estimates of OFG will be significantly revised if active margins, with steep coastal topographies like the Canterbury margin, are considered.

The concept of green chemistry has made significant impact on many frontages including the use of green solvents or sustainable catalyst materials. Benzimidazole ring is an important nitrogen-containing heterocyclic, which exhibits a broad spectrum of bioactivities and are widely utilized by the medicinal chemists for drug discovery. A simple and efficient method was developed for the synthesis of some benzimidazole derivatives via reaction of o -phenylenediamine and substituted aldehydes in the presence of nano-SnCl 4 /SiO 2 as a mild catalyst. Ten 2-substituted benzimidazole compounds ( J 1 -J 10 ) were synthesized. All compounds were evaluated against different species of yeasts and filament fungi using broth micro dilution method as recommended by clinical and laboratory standard institute. Among these compounds, the active ones were chosen for their cytotoxic activities evaluation against MCF-7 and A549 cell lines using MTT method. Compound J 2 showed the best antifungal activity against all tested species. Compounds J 5 -J 7 had also desirable antifungal activities. Our cytotoxic results were also similar to the antifungal activities except for J 7 which had no cytotoxic activity., (Copyright: © 2019 Research in Pharmaceutical Sciences.)

Background: NK (natural killer) and NKT (natural killer T) cells, as components of innate immune system, play a crucial role in tumor progression and dissemination., Objective: To investigate the percentages of NK cells, NKT cells, iNKT (invariant natural killer T) cells, total T lymphocytes as well as activated T lymphocytes, in tumor draining lymph nodes (TDLNs) of patients with breast cancer (BC) and their association with different clinic-pathological features of the patients., Methods: Axillary lymph nodes were obtained from 30 Iranian women with breast cancer. After routine pathological evaluations, mononuclear cells were separated from their lymph nodes and incubated with appropriate fluorochrome conjugated monoclonal antibodies specific for CD3, HLA-DR, CD16/56, and Vα24Jα18-TCR. Data were collected on a four-color flow cytometer and analyzed by CellQuest software., Results: The mean percentages of NK (CD3-CD16/56+), NKT (CD3+CD16/56+) and iNKT (Vα24Jα18-TCR+) cells in TDLNs mononuclear cells of BC patients were 2.04%, 2.44% and 0.1%, respectively. A significant decrease in the percentages of NK and iNKT subsets in patients with grade I was observed compared to grade III (p=0.03 and p=0.01, respectively). Moreover, NK cells were increased in patients with grade III of BC compared to grade II (p= 0.003)., Conclusion: The increase in the percentage of NK and iNKT cells in TDLNs of patients with higher grade of BC might suggest a suppressive phenotype for these cells in breast cancer, which merit more functional investigation.

Background: A T cell subtype with the CD4 + CD25 - FoxP3 + phenotype was recently described. We aimed to investigate the frequency of these cells and their ability to produce cytokines in tumor-draining lymph nodes from patients with breast cancer (BC)., Materials and Methods: Mononuclear cells from lymph nodes of 20 patients with BC were activated and stained for appropriate markers. The cells were assayed with four-color flow cytometry., Results: A very small fraction of CD4 + CD25 - FoxP3 + cells produced cytokines at levels that were significantly lower than in the regulatory (CD4 + CD25 + FoxP3 + ) and effector cell (CD4 + CD25 + FoxP3 - ) subpopulations. The expression of IFNγ and IL-2 in the CD4 + CD25 - FoxP3 + subset was significantly higher than in Treg cells, but lower than in the effector subset. Conversely, IL-22 expression in Treg cells was significantly higher than in the CD4 + CD25 - FoxP3 + subpopulation. The expression of IL-10 in the CD4 + CD25 - FoxP3 + subset was also significantly higher than in effector cells., Conclusion: We suggest that CD4 + CD25 - FoxP3 + cells in patients with BC are exhausted cells with an intermediate phenotype between effector and regulatory cells., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Head and neck squamous cell carcinoma (HNSCC) is the sixth leading incident cancer worldwide. In this study, we aimed to investigate the possible association of OX40 gene polymorphisms, rs17568G/A and rs229811A/C, with susceptibility to HNSCC and its clinicopathological features. Two hundred and two HNSCC patients and 200 healthy age-sex matched individuals were enrolled. rs17568G/A and rs229811A/C polymorphisms in OX40 gene were genotyped using RFLP-PCR method. We observed more than 2 times increased risk for squamous cell carcinoma development in nose and paranasal sinuses among individuals who inherited GG genotype at rs17568 region (OR 2.29; CI 1.01-5.20; P = 0.035). Considering rs2298211 SNP, AA genotype was also observed with higher frequency, in comparison with other two genotypes (AC or CC), among patients with HNSCC originated from these regions (P = 0.003). Besides, we observed that patients with C allele at this locus (AC and CC genotypes) have tumors with significantly higher histological grade (P = 0.042). Our findings suggest the possible association of rs17568 GG genotype, as well as rs2298211 AA genotype with susceptibility to develop squamous cell carcinoma in the nose and sinonasal cavities.

Purpose: In recent years, the prognostic/predictive significance of tumor infiltrating lymphocytes (TILs) has become a topic of interest. Here, we aimed to evaluate the prognostic significance of CD3+, CD8+, CD45RO+ and Foxp3+ TILs in breast cancer, as well as the relation of these markers to other clinicopathological features of this disease., Methods: FFPE tumor samples from 94 females with invasive ductal carcinoma of the breast were retrospectively selected and immunohistochemically assessed for CD3, CD8, CD45RO and Foxp3 expression. Digital photos were acquired from the center (CT) and invasive margins (IM) of the tumors, after which positive cells were counted using ImageJ software., Results: We found that greater infiltrations of target lymphocyte subpopulations were associated with TNM stage III, lymph node metastasis, high histological grade, ER negativity and HER2 positivity. The ratios of CD8+ cytotoxic T cells to CD3+, CD45RO+ and Foxp3+ TILs were found to be relatively higher in tumors exhibiting the aforementioned characteristics. In univariate survival analyses, CD8+ TILs in the IM and total CD45RO+ TILs were found to be significantly associated with overall survival (OS). Infiltration of CD45RO+ TILs in the CT and lymph node status were variables that significantly correlated with disease-free survival (DFS). Multiple Cox regression analyses revealed independent significant prognostic effects of total CD45RO+ TILs and lymph node status (HR of 3.24 and 3.19, respectively) in predicting OS. Infiltration of CD45RO+ TILs in the CT (HR 3.12) and lymph node status (HR 3.15) also exhibited significant prognostic effects on DFS., Conclusion: From our data we conclude that CD45RO+ TILs serve as prognostic factors for predicting OS and DFS of breast cancer patients.

Purpose: The role of cytokine-producing B cells in antitumor immunity is mostly overlooked. In the present study, we investigated changes in B cell cytokine profiles in breast tumor-draining lymph nodes (TDLNs) during disease progression, and associations of these changes with prognostic indicators., Methods: Flow cytometry was used to measure the expression of TNF-α, IL-10, TGF-β, IL-2 and IFN-γ in B cells from 42 axillary lymph nodes. The frequencies of IL-10 + and FoxP3 + T regulatory cells (Tregs) were also determined., Results: No significant changes in B cell cytokine profiles were observed during breast cancer progression from stage I to III, but the percentage of B cells with high TNF-α expression (TNF hi ) showed a negative relationship with lymph node involvement and Her2 expression (p < 0.05). The percentage of IL-10 + B cells was found to be significantly higher in non-metastatic lymph nodes in node-negative compared to node-positive patients (p = 0.001). The frequencies of IL-10 + and TNF hi B cells were found to be negatively correlated with the number of involved lymph nodes. The frequency of TNF hi B cells showed an inverse correlation with the frequency of FoxP3 + Tregs, which in turn was associated with indicators of a poor prognosis., Conclusions: Our data indicate that the cytokine profiles of B cells in TDLNs of patients with breast cancer show associations with various disease parameters. TNF hi and IL-10 + B cells correlated positively with indicators of a good prognosis. Further functional studies are required to elucidate the role of cytokine production by B cells in immunity against breast cancer.

Recently the quinazoline derivatives have attracted much attention for their anticancer properties. In this study a series of new brominated quinazoline derivatives ( 1a-1g ) were synthesized in two steps. In the first step we used N-bromosuccinimide to brominate the anthranilamid. Then in the second step we closed the quinazoline ring by different aromatic aldehydes. Our aldehydes contain different electron donating or electron withdrawing groups at different positions of the aromatic ring. The chemical structures of products were confirmed by spectroscopic methods such as IR, 1HNMR, 13CNMR, and mass spectroscopy. The cytotoxic activities of the compounds were assessed on three cancerous cell lines including MCF-7, A549, and SKOV3 using colorimetric MTT cytotoxic assay in comparison with cisplatin as a standard drug. Our results collectively indicated that 1f and 1g , exhibited the best anti-proliferative activities on three investigated cancerous cell lines., (Copyright: © 2019 Research in Pharmaceutical Sciences.)

Introduction: Head and neck squamous cell carcinomas (HNSCCs) are the most common cancers of head and neck and the sixth most common malignancy worldwide. Programmed cell death 1 (PD-1) is an immune inhibitory molecule which through interaction with its ligands recruits protein phosphatase resulting in immune response inhibition. Expression of PD-1 ligands on tumor cells is considered as one of the crucial immune evasion mechanisms. This study aimed to investigate the association of PD-1 gene polymorphisms at positions PD1.3 (rs11568821), PD1.5 (rs2227981) and PD1.9 (rs2227982) with susceptibility to HNSCCs., Subjects and Methods: 150 patients pathologically confirmed to suffer from HNSCCs and 150 age-sex matched healthy controls were recruited in this study. Genomic DNA was extracted from white blood cells of all participants. Restricted fragment length polymorphisms (RFLP)-PCR was performed using site specific primers to determine the genotypes in each position., Results: Statistical analyses indicated no significant differences in the frequencies of genotypes, alleles as well as haplotypes between patients and controls (P > 0.05), however, haplotype combination differed significantly between two groups. GCC/GCT, GCC/GCC and GCT/GCC were higher in the HNSCC patients than the control group (P < 0.05). On the other hand, in the controls, GCT/GCT, GCT/ACC, GCT/ACT and ACC/GCT were more frequent. No significant association was found with various HNSCC clinicopathological characteristics., Discussion: Our results suggested that although PD-1 gene polymorphisms at three investigated positions are not solely associated with susceptibility to HNSCCs, haplotype combinations emerged from these three loci may render susceptibility.

Breast cancer, as the most common cancer in women worldwide, represents about 30% of all cancers affecting women. Long non-coding RNAs (lncRNAs) have been implicated in the regulation of several biological processes, and their dysregulation in cancer has well been documented. To investigate possible age-dependent variations in expression profiles of lncRNAs, we evaluated the expression levels of four lncRNAs, i.e., MALAT1, GAS5, SRA, and NEAT1, in breast cancer (BC) samples obtained from younger (<45 years) and older (>45 years) women. Tumor samples (n = 23) and 15 normal tissues were collected from BC patients. All tumor and normal samples were morphologically confirmed by a pathologist. RNA was extracted from the tissues and cDNAs were then synthesized. The lncRNA expression levels were evaluated by qRT-PCR. The changes in the expression levels were determined using the ΔΔCt method. Compared to normal tissues, BC tissues from both age groups (women under 45 years of age and women above 45 years of age) showed upregulation of MALAT1 (p = 0.003 and p = 0.0002), SRA (p = 0.005 and p = 0.0002), and NEAT1 (p = 0.010 and p = 0.0002) and downregulation of GAS5 (p = 0.0002 and p = 0.0005). Additionally, our analysis showed significant and direct correlation between the age and the expression levels of three of the four lncRNAs studied in this work. All four lncRNAs were overexpressed in both MDA-MB-231 and MCF7 cell lines (p = 0.1000). Our data show that MALAT1, GAS5, SRA, and NEAT1 lncRNAs are dysregulated in BC samples. However, except for MALAT1, the expression levels of all of these lncRNAs were significantly lower in cancers developed in younger cases, where poorer prognosis is suggested. Of note, GAS5 reduced expression has been documented to correlate with tumor progression., (Copyright © 2018. Published by Elsevier Inc.)

Background: The compartments of memory T cells play a fundamental role in the immune system by substantiating specific and acquired immunity. A new subset of memory cells, T stem cell memory (T SCM ) cells, with stem cell-like properties, a high capacity to proliferate, a long survival, and an ability to differentiate into all effector and memory cells has recently been introduced. In the present study, we aimed to determine the frequency of CD4 + T SCM and other T memory cell subsets in tumor draining lymph nodes of breast cancer patients., Materials and Methods: Mononuclear cells were obtained from axillary lymph nodes of 52 untreated patients with breast cancer (BC) and stained with fluorochrome conjugated anti-CD4, -CCR7, -CD45RO and -CD95 antibodies to detect different subtypes of memory cells in CD4 + lymphocyte populations. Data were acquired using a four-color FACSCalibur flow cytometer and analyzed using CellQuest Pro software., Results: We found that >70% of CD4 + lymphocytes in draining lymph nodes of BC patients exhibited a memory phenotype of which 7.04 ± 1.04% had a T SCM phenotype (CD4 + CCR7 + CD45RO - CD95 + ). The frequency of T SCM cells was significantly higher in tumor positive lymph nodes compared to tumor negative lymph nodes (p = 0.026) as well as among those patients who had at least one affected lymph node (p = 0.012). Moreover, we found that the total frequency of central memory T cells (T CM ) with a low expression of CD45RO was significantly higher among these patients. The percentage of CD45RO Low T CM cells was also found to increase with tumor progression from stage I to stage III (p = 0.020). On the other hand, we found that the percentage of CD95 Hi effector memory T cells (T EM ) was significantly decreased in involved lymph nodes (p = 0.009)., Conclusion: Our data suggest that following long-term exposure to putative tumor antigens, T SCM cells proliferate to generate a pool of committed memory and effector T cells. As the tumor progresses, the immunosuppressive milieu induced by tumor cells may slow down the differentiation of CD45RO Low T CM cells to more functional sub-populations.

To find out if the T cell repertoire is efficiently and specifically provoked in patients with breast cancer, we have investigated the clonotypes of main T cell subsets (based on Vβ-Chain) in tumor draining lymph nodes. CD4+ helper, CD8+ cytotoxic and (CD4+CD127dimCD25+) regulatory T cells, were negatively selected, and isolated, from lymph node mononuclear cells of 14 untreated patients with breast cancer. Four non-malignant patients, who underwent surgical operation, were also recruited as the control group. Based on sequences and new nomenclature of the T cell Receptor β Variables (TRBVs) available in the international ImMunoGeneTics (IMGT) database, 28 TRBV specific forward primers and two TRB Constant region (TRBC) specific reverse primers were developed to amplify all functional alleles. Fluorescent-labeled PCR products were then run on an ABI PRISM 310 Genetic-Analyzer. The data was analyzed by GeneMapper software version 3.1. Clonotype analysis suggested that activated T cells are present in breast cancer. More TRBV usage were detected among CD4+ helper and regulatory subsets, with Gaussian-like pattern in the majority of functional TRBV families; whereas CD8+ cytotoxic T cells showed oligoclonality in almost all TRBV families with one or two dominant peaks in each family. Similar pattern in some of these TRBVs were also observed among controls. Having no expression or polyclonality in the controls, the oligoclonal pattern observed in the TRBV18, however, appears to be specific to breast cancer patients. This phenomenon may reflect the existence of new antigenic stimulation(s) in BC patients, preferentially activating those clones of T cells that express TRBV18.

Dichloroacetate (DCA) is a simple and small anticancer drug that arouses the activity of the enzyme pyruvate dehydrogenase (PDH) through inhibition of the enzyme pyruvate dehydrogenase kinases (PDK1-4). DCA can selectively promote mitochondria-regulated apoptosis, depolarizing the hyperpolarized inner mitochondrial membrane potential to normal levels, inhibit tumor growth and reduce proliferation by shifting the glucose metabolism in cancer cells from anaerobic to aerobic glycolysis. In this study, a series of DCA analogues were applied to quantitative structure-activity relationship (QSAR) analysis. A collection of chemometrics methods such as multiple linear regression (MLR), factor analysis-based multiple linear regression (FA-MLR), principal component regression (PCR), and partial least squared combined with genetic algorithm for variable selection (GA-PLS) were applied to make relations between structural characteristics and cytotoxic activities of a variety of DCA analogues. The best multiple linear regression equation was obtained from genetic algorithms partial least squares, which predict 90% of variances. Based on the resulted model, an in silico -screening study was also conducted and new potent lead compounds based on new structural patterns were designed. Molecular docking as well as protein ligand interaction fingerprints (PLIF) studies of these compounds were also investigated and encouraging results were acquired. There was a good correlation between QSAR and docking results.

Cyclometalated rollover complexes of the type [PtMe(κ 2 N,C-bipyO-H)(L)] [bipyO-H=cyclometalated 2,2'-bipyridine N-oxide; L=tricyclohexylphosphine (PCy 3 , 2 a), 2-(diphenylphosphino)pyridine (PPh 2 py, 2 b), P(OPh) 3 (2 c)] were synthesized by treating [PtMe(κ 2 N,C-bipyO-H)(SMe 2 )] (1) with various monodentate phosphine and phosphite ligands. These complexes were characterized by NMR spectroscopy, and the structure of 2 a was confirmed by single-crystal X-ray diffraction. Complex 1 was treated with bis(diphenylphosphino)methane (dppm) at a 1:1 ratio to give the corresponding [PtMe(κ 2 N,C-bipyO-H)(κ 1 P-dppm)] (3 b) complex, in which the dppm ligand acts as a monodentate pendant ligand. The biological activities of these complexes were evaluated against a panel of four standard cancer cell lines: lung carcinoma (A549), ovarian carcinoma (OV-90 and SKOV3), and breast carcinoma (MCF-7). Complexes 2 c and especially 3 b indicated effective potent cytotoxic activity regarding the cell lines. Electrophoresis mobility shift assays and molecular-modeling investigations were performed to determine the specific binding mode and the binding orientation of these alkylating agents to DNA. Detection of cellular reactive oxygen species was also determined., (© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Aromatase inhibitors (AIs) as effective candidates have been used in the treatment of hormone-dependent breast cancer. In this study, we have proposed 300 structures as potential AIs and filtered them by Lipinski's rule of five using DrugLito software. Subsequently, they were subjected to docking simulation studies to select the top 20 compounds based on their Gibbs free energy changes and also to perform more studies on the protein-ligand interaction fingerprint by AuposSOM software. In this stage, anastrozole and letrozole were used as positive control to compare their interaction fingerprint patterns with our proposed structures. Finally, based on the binding energy values, one active structure (ligand 15) was selected for molecular dynamic simulation in order to get information for the binding mode of these ligands within the enzyme cavity. The triazole of ligand 15 pointed to HEM group in aromatase active site and coordinated to Fe of HEM through its N4 atom. In addition, two π-cation interactions was also observed, one interaction between triazole and porphyrin of HEM group, and the other was 4-chloro phenyl moiety of this ligand with Arg115 residue.

Background: Traditional approaches in Continuing Medical Education (CME) appear to be ineffective in any improvement of the patients' care, reducing the medical errors, and/or altering physicians' behaviors. However, they are still executed by the CME providers, and are popular among the majority of the physicians. In this study, we aimed to explore the parameters involved in the degree of effectiveness of CME program in Iran. Methods: In this study, 31 participants, consisting of general practitioners, CME experts and providers were recruited to participate in in-depth interviews and field observations concerning experiences with CME. Application was made of the qualitative paradigm along with the qualitative content analysis, using grounded theory data analysis methodology (constant comparative analysis). Results: Based on the participants' experiences, the insufficient consistency between the training program contents and the demands of GPs, in addition to the non-beneficiary programs for the physicians and the non-comprehensive educational designs, created a negative attitude to the continuing education among physicians. This could be defined by an unrealistic continuing education program, which is the main theme here. Conclusion: Impracticable continuing education has created a negative attitude toward the CME programs among physicians so much that they consider these programs less important, resulting in attending the said programs without any specific aim: they dodge absenteeism just to get the credit points. Evidently, promoting CME programs to improve the performance of the physicians requires factual needs assessment over and above adaptation of the contents to the physicians' performance.

Background: Toll-like receptor 9 (TLR9) is a DNA receptor of innate immune system which plays a pivotal role in inflammatory response. Recent evidence reveals over-expression and functionality of TLR9 in a wide variety of cancer cells and its contribution to tumor cell proliferation and survival., Objective: In this study, we assessed the aberrant cell surface expression of TLR9 in cancer using cell-lines model., Methods: Three breast cancer cell-lines (MDA-MB-231, MCF7 and SKBR3) and five colorectal adenocarcinoma cell-lines (HT29, HT29/219, SW480, SW48 and SW1116) in addition to one primary foreskin isolated fibroblast cell were analyzed for cell surface and intracellular expression of TLR9 by flow cytometry method., Results: Maximum surface expression of TLR9 was observed in colorectal cell-line HT29/219 (38.35%), as compared with the bottom line fibroblast normal cells (0.12%). The most intracellular expression was observed in MCF-7 cells (35.63%), whereas MDA-MB-231 expressed the maximum surface/intra cellular expression (277 times)., Conclusions: Based on the results, we hypothesize that aberrant surface expression of TLR9 on tumor cells may promote tumor growth and invasion. It might also highlight a dual contradictory role for CpG-ODNs, as adjutant in cancer therapy.

Background: Cytotoxic CD8+ T cells, as essential parts of the adaptive immune system, play pivotal roles in anti-tumor immune responses. It is well documented that cytokine expression profiles and activation status of these cells during anti-tumor immune responses affect the outcome of host-tumor interaction., Objective: To investigate the percentages of CD8+ lymphocytes and their subsets in tumor draining lymph nodes of patients with bladder cancer., Methods: Forty-five patients with bladder cancer, candidate for radical cystectomy, were recruited. Mononuclear cells were isolated from draining lymph nodes using Ficoll-Hypaque gradient centrifugation, and were activated by PMA/Ionomycin in the presence of Golgi inhibitors. The cells were then permeabilized and stained with appropriate flourochrome conjugated antibodies against CD3, CD8, IFN-γ, IL-17 and IL-4 molecules. Data were collected on a four-color flow cytometer and analyzed by CellQuestPro software., Results: Despite no difference in the frequency of IL-17 producing CD8+ (Tc17) lymphocytes, the mean expression of IL-17 in this subset was significantly elevated in high-grade patients (p=0.011). The percentage of double positive IFN-γ/IL-17 CD8+ lymphocytes was also significantly increased in node positive patients compared to node negative ones (p=0.046). Our results also demonstrated that the percentage of IFN-γ producing CD8+ (Tc1) lymphocytes was significantly increased in the patients with higher histological grade compared to those with lower ones (p=0.038)., Conclusion: IFN-γ and IL-17 producing CD8+ T cells may increase in advanced stages of bladder cancer, but their correlation with tumor prognosis remains to be investigated.

Introduction: Continuing Medical Education (CME) has been considered as a lifelong commitment for doctors to provide the optimal care for patients. Despite a long history of creating CME programs, outcomes are far from ideal. The present qualitative study aims to clarify the barriers affecting effectiveness of the CME programs in Iran based on the experiences of general practitioners., Methods: Sixteen general practitioners were recruited to participate in in-depth interviews and field observations concerning experiences with CME. The study was performed using a qualitative content analysis method. The codes, categories and themes were explored through an inductive process in which the researchers moved from specific to general., Results: The participants' experiences identified a number of barriers, particularly insufficient interaction with the instructors; additional problems included the teachers' use of an undifferentiated approach; unreal and abstract CME; and ignorance of the diverse reasons to participate in CME., Conclusion: Based on the study results, there are multiple barriers to effective implementation of CME in Iran. The key barriers include insufficient interaction between the trainees and providers, which must be considered by other stakeholders and program designers. Such interactions would facilitate improved program design, invite more specific tailoring of the education to the participants, allow for more effective educational methods and set the stage for outcome evaluation from the learners actually applying their new knowledge in practice. Replication of these findings with another sample would improve confidence in these recommendations, but these findings are broadly consistent with findings in the educational literature on improving the efficacy of CME.

The relationship between salivary gland tumors and immune system has not been well inspected. We aimed to investigate the distribution of CD4(+)CD25(+)Foxp3(+) regulatory T (Treg) cells, CTLA4(+)CD4(+) lymphocytes, as well asIL-17 producing CD4(+) and CD8(+) (Th17 and Tc17) lymphocytes in peripheral blood of patients with benign and malignant salivary gland tumors and a group of healthy controls. Peripheral blood samples were obtained from 27 patients with salivary gland tumors (19 benign and 8 malignant; mean age of 49.2±18.3), as well as19 age/sex matched healthy donors. Fluorochrome-conjugated antibodies were used to stain the cell surface markers, as well as intracellular molecules following cell-membrane fixation and permeabilization. The stained cells were acquired on a FACSCalibur four-color flowcytometer and analyzed by CellQuest Pro software package. The data were presented as mean percentages±SEM. Results indicated that the patients with malignant salivary gland tumors have increased percentage of Treg cells (7.74±1.1) and intracellular CTLA4 (inCTLA4)-positive CD4(+) lymphocytes (8.18±1.77) in comparison to the patients with benign tumors (4.38±0.56 for Treg cells and 3.83±0.56 for CTLA4(+)CD4(+) cells), as well as control subjects (2.34±0.28 for Treg cells and 2.22±0.25 for CTLA4(+)CD4(+) cells) (p≤0.001). Conversely these patients had reduced percentage of Th17 cells (0.84±0.14) comparing to the patients with benign tumors (2.09±0.31) as well as control subjects (2.31±0.23) (p≤0.001). In addition, the ratio of Th17/Treg lymphocytes was significantly lower in both malignant (0.12±0.03) and benign (0.48±0.09) tumors in comparison to control subjects (1.26±0.23) (p<0.001). The mean percentage of Tc17 cells in patients with benign (1.14±0.15) and malignant (0.60±0.13) tumors was nearly similar to those in control subjects (0.83±0.14) but the mean expression intensityofIL-17 by these cells was significantly higher in patients with malignant tumors (11.06±1.26) than controls (7.61±0.69) (p=0.01). Increase in the prevalence of regulatory lymphocytes, Treg cells and CTLA4(+)CD4(+) lymphocytes, as well as the imbalance of Th17/Treg ratio may suggest the contribution of these immune effector cells in the progression of salivary gland tumors. From immune-regulatory point of view, these data also suggest that benign salivary gland tumors might fall between healthy and malignant conditions. The immunity to salivary gland tumors, as well as the findings presented here merits more in-depth investigation., (Copyright © 2015. Published by Elsevier B.V.)

Tumor draining lymph nodes form the first line of defense against tumor dissemination. Lymphocyte subpopulations activated during anti-tumor response determine the outcome of host-tumor interaction. In the present study we explored the percentages of different subtypes of CD4+ lymphocytes, including regulatory cells (TFR, CD25-, and CD25+ Treg cells), helper subsets (Th1, Th2, Th17, and Tfh cells), and the expression level of their cognate cytokines (IFNγ, IL4, and IL17) in tumor draining lymph nodes of patients with breast cancer, and compared the results between node negative (LN-) and node positive (LN+) patients. Forty seven sentinel and non-sentinel auxiliary lymph nodes with or without tumor involvement were collected from untreated breast cancer patients undergoing surgical resection. Mononuclear cells obtained from fresh homogenized lymph nodes were subjected to surface and intracellular staining by flow cytometry. The results revealed the presence of a newly identified subtype of regulatory T cells, TFR, as well as CD25- Treg cells in TDLNs of the breast cancer patients. In addition, evaluation of different helper and regulatory subgroups of CD4+ T lymphocytes showed that upon metastasis of tumor cells to lymph nodes together with the progression of the disease stage, the immune responses changed from an inflammatory to an inhibitory state, as evidenced by a reduction in pro-inflammatory and anti-tumor cytokines, IL17 and IFNγ, as well as an increase in pro-tumorigenic phenotypes, Th2 and Treg cells. This situation may provide a favorable condition for tumor growth and spread., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background: CD8+ cytotoxic T lymphocytes have been recently divided based on their cytokine expression profile., Objective: To evaluate the percentages of CD8+ lymphocytes and their effector subsets including Tc1, Tc2 and Tc17 in the tumor draining lymph nodes (TDLNs) of patients with breast cancer., Methods: Single cell suspensions were obtained from TDLNs of 42 patients with breast cancer. Staining of the cell surface markers and intracellular cytokines was performed using appropriate fluorochrome-conjugated antibodies. The data was acquired on a four-color flow cytometer and was analyzed by CellQuestPro software package. The percentages of different CD8+ cell subtypes (Tc1, Tc2 and Tc17) were quantified in CD8+ T lymphocytes. The comparison was made between LN+ versus LN- patients, as well as patients in different clinico-pathological status., Results: The percentage of Tc1, Tc2 and Tc17 subsets were not significantly different between LN+ and LN- patients. Despite no difference in the percentages of Tc1 cells in LN+ patients with infiltrative ductal carcinoma (IDC), the mean expression of IFN-γ by Tc1 cells decreased significantly in comparison to LN- patients. On the other hand, the percentages of Tc2 and Tc17 effector subsets were increased in advanced stages (p=0.018 and p=0.009, respectively)., Conclusion: As the first study to investigate various effector subtypes of CD8+ lymphocytes in TDLNs of patients with breast cancer, our data collectively suggests a positive association between IL-17- and IL-4-producing CD8+ T cell percentages (Tc2 and Tc17) in TDLNs with breast cancer progression. Although the number of Tc1 cells seems not to be affected by cancer progression, down-regulation of IFN-γ by these cells seems to be associated with tumor metastasis to TDLNs. These findings may have implications in cancer immunotherapy based on CD8+ effector subsets.

Background: Osteoarthritis (OA) is the most common form of arthritis seen clinically. Current treatments for OA are limited to decreasing associated pain, maintaining or improving joint function, and minimizing disability. However, these treatments have no effect on the regeneration of hyaline cartilage. Since mesenchymal stem cells (MSCs) have been described as promising cell sources for cartilage repair, the present study was designed to examine whether intra-articular injection of scaffold-free adipose-derived stem cells (ASCs) obtained from subcutaneous adipose tissue could restore the matrix of arthritic knee joints in mature animals., Methods: OA was induced in adult white New Zealand rabbits by unilateral anterior cruciate ligament transection (ACLT); the contralateral knee was considered the sham-operated group. At 12 weeks following surgery, the ASCs treated group was injected intra-articularly with a single dose of 1 × 10(6) cells suspended in 1 mL of medium. The control group received 1 mL of medium without cells and the sham-operated group received no treatment. All rabbits were sacrificed at 16 and 20 weeks after surgery. OA progression was evaluated radiologically, grossly, and histologically using hematoxylin and eosin, Safranin-O, and toluidine blue staining., Results: At 12 weeks after surgery all knees subjected to ACLT showed radiological signs of OA. The findings showed significant differences in the quality of cartilage between ASCs-injected group compared to control group, particularly at 20 weeks after surgery., Conclusion: This study suggests that ASCs obtained from subcutaneous adipose tissue could be a viable approach for treating OA.

Interleukin-13 (IL-13) is a TH2 cytokine with direct and indirect immunoregulatory functions on cancer cells. The cytokine has been reported to have some polymorphic variations at the gene level associated with some immune related diseases including asthma and allergy. In the present study, association of three IL13 gene polymorphisms at positions -1512 A/C and -1055 C/T in the promoter and +2044 G/A in exon-4 was investigated in Iranian women with breast cancer and healthy controls. Genotyping of IL13 gene polymorphisms were performed by PCR-RFLP methods. Serum level of IL-13 was assessed by ELISA. Haplotypes were constructed from genotypic data using Arlequin 3.1 software package. Haplotype analysis revealed higher frequency of a three-locus haplotype, ACA (-1512A/-1055C/+2044A), in normal women than breast cancer patients (P < 0.025). Haplotype CCA, from the other hand, was observed with more frequency among patients than controls (P < 0.03). No statistically significant differences were found in the frequency of genotypes and alleles between patients and control group. No association was observed between investigated genotypes and other prognostic factors including tumor type, lymph node involvement and tumor size. IL-13 serum level was undetectable in both patients and control subjects. Despite observing no association between breast cancer and the single SNPs, results of this investigation suggest that the presence of CCA haplotype of IL13 gene may be associated with susceptibility of Iranian women to breast cancer.

Lung cancer is one of the leading causes of death from cancer. Both immune cells and tumor cells play a key role in lung cancer immunity by secretion of cytokines and developing type-2 cell-mediated immune response. IL-13 is an immunoregulatory cytokine affecting tumor immunosurveillance by deviation of immune response from Th1 to Th2. In the present study we sought to determine the association of single nucleotide polymorphisms (SNPs) of IL-13 gene at positions +2044 (G/A) and -1055 (C/T) and lung cancer. One hundred forty one patients and 113 controls were recruited; control group was subdivided into smoker and nonsmoker individuals for serum detection. Genotyping was carried out by PCR-RFLP assay and IL-13 detection by ELISA method. No statistically significant difference was found in the frequency of genotypes, alleles, and haplotypes at positions +2044 (G/A) and -1055 (C/T) of IL-13 gene between lung cancer patients and controls. Serum level of IL-13 was not detectable in both groups. The results of this study reveal that although +2044 (G/A) and -1055 (C/T) SNPs in IL-13 are implicated in some pulmonary processes, they do not confer susceptibility to lung cancer in Iranian population.