Lung cancer (LC), which includes small-cell lung carcinoma (SCLC) and non-small-cell lung carcinoma (NSCLC), is common and has a high fatality rate. This study aimed to reveal the prognostic mechanisms of LC. GSE30219 was extracted from the Gene Expression Omnibus (GEO) database, and included 293 LC samples and 14 normal lung samples. Differentially expressed genes (DEGs) were identified using the Limma package, and subjected to pathway enrichment analysis using DAVID. MicroRNAs (miRNAs) targeting the DEGs were predicted using Webgestalt. Cytoscape software was used to build a protein-protein interaction (PPI) network and to identify significant network modules. Survival analysis was conducted using Survminer and Survival packages, and validation was performed using The Cancer Genome Atlas (TCGA) dataset. The good and poor prognosis groups contained 518 DEGs. miR-190, miR-493, and miR-218 for the upregulated genes and miR-302, miR-200, and miR-26 for the downregulated genes were predicted. Three network modules (module 1, 2, and 3) were identified from the PPI network. CDK1, MCM10, and NDC80 were the core nodes of module 1, 2, and 3, respectively. In module 1, CDK1 interacted with both CCNB1 and CCNB2. Additionally, CDK1, CCNB1, CCNB2, MCM10, and NDC80 expression levels correlated with clinical survival and were identified as DEGs in both GSE30219 and the TCGA dataset. miR-190, miR-493, miR-218, miR-200, and miR-302 might act in LC by targeting the DEGs. CDK1, CCNB1, CCNB2, MCM10, and NDC80 might also influence the prognosis of LC.
Background: Oxidative damage is of great importance for patients with breast cancer. Thus, studies were performed to identify the relationship between breast cancer and oxidative stress biomarkers.
Objectives: In this study, our aim was to find out the oxidative and antioxidant status, serum thiol-disulphide levels in subjects with breast cancer.
Methods: This study was conducted between March and June 2018 with 82 control subjects (aged between 32-67 years) and 127 breast cancer patients (aged between 27-66 years) (p=0.058) in Selcuk University Faculty of Medicine, Konya, Turkey. Serum myeloperoxidase (MPO), catalase, prolidase were analyzed with kinetic spectrophotometric and thiol-disulphide, ischemia-modified albumin (IMA), ceruloplasmin were detected by colorimetric methods.
Results: Serum levels of catalase [199.3 (16.4-489.9) vs 81.6 (18.2-322.9) (kU/L)], MPO [124±28 vs 101±31 U/L], disulphide [25 (11-61) vs 18 (2-41) µmol/L], IMA [0.66 (0.31-3.30) vs 0.62 (0.19-1.31) absorbance unit (ABSU)] and prolidase levels [2217±538 vs 1456±401 U/L] were higher in patients than control subjects (For all p<0.001 except for IMA p=0.031). Native thiol [342±60 vs 391±52 µmol/L] and total thiol levels [396±56 vs 430±52 µmol/L] were lower in patients compared with the control group (For all p<0.001).
Conclusions: Levels of serum thiol/disulphide and prolidase might be reliable indicators for determining oxidative status in certain patient populations.
Purpose: The implantable bladder cancer (BC) models allow the researchers to perform rapid and useful experiments for BC. We investigated the implantation success of BC cells obtained from Wistar rats (grown in vitro), into bladders of syngeneic Wistar rats, which are commonly used in the laboratories.
Methods: The Nara Bladder Tumor No.2 (NBT-II) BC cells induced with 4-hydroxybutylnitrosamine were grown with passages in Kocaeli University Center for Stem-Cell and Gene-Therapies. After urothelial denudation, 2x106 NBT-II cells were then implanted into bladders of 24 female Wistar rats (aged 7-8 weeks). The rats were randomly divided into four experimental groups; three instillation groups (8 per group) and one sham-operated control group consisting of 6 rats. First, second and third instillation groups were sacrificed at days 7, 14, and 21, respectively, and, bladders were histopathologically evaluated for BC according to WHO / International Society of Urological Pathology.
Results: All tumors were pT1 (including 1 rat that prematurely died at 5th day), except one rat that died prematurely at 8th day had pT2 tumor. Implantation rates were 28.58% (2/7) in the first group, and 42.85% (3/7) in the second, for a cumulative rate of 35.71% (5/14) in these two-groups (until 14th day). Interestingly, there was no tumor in the third group, but there was an inflammatory granulation tissue.
Conclusion: Seeding NBT-II cells into bladders of Wistar rats was described, successfully tested and demonstrated in this study. This implantable BC model of Wistar rats may be improved to increase the success rate of BC cell implantation in new studies with higher number of animals.
Purpose: Ovarian torsion, being a gynecological emergency, requires to be rapidly diagnosed and treated with minimal consequences on ovarian function after the removal of torsion. As ischemia modified albumin (IMA) is considered a good biomarker in diverse ischemic diseases, the aim of our study was to determine the effect of different ovarian torsion/detorsion models on serum and ovarian homogenates levels of IMA in an experimental study.
Methods: IMA was measured in the serum and ovarian homogenates of 7 groups of female rats (10 animals in each group): 1 - control (no intervention); 2 - sham (only laparotomy); 3 - ischemia group: 3 hours ovarian torsion (OT); 4 - 3 hours OT (ischemia), 1 hour simple reperfusion; 5 - 3 hours ischemia, 1 hour controlled reperfusion that was assured during the first two minutes by opening and closing the clips on the ovarian pedicles in 10 seconds intervals, followed by simple reperfusion; 6 - 3 hours ischemia, 24 hours simple reperfusion; 7 - 3 hours OT, 24 hours controlled reperfusion. The results were analyzed by Welch’s ANOVA and Spearman correlation.
Results: Ischemia increases the IMA in both serum and ovarian homogenates compared to control and sham groups. The controlled reperfusion groups had a statistically significant lower IMA in serum compared to simple reperfusion groups. IMA was found to be higher in the ovarian homogenates of simple reperfusion compared to controlled reperfusion groups.
Conclusion: Our results suggest that controlled reperfusion prevent the processes that increase the IMA in ovarian torsion.
Introduction: Experimental acute cerebral ischemia quickly triggers circulating inflammatory cells, provoking infiltration of neutrophils and macrophages in the damaged brain region. N-3 polyunsaturated fatty acids alleviate the ischemic deterioration, however, their potential effect on bone marrow cell mobilization is less known.
Materials and methods: healthy male Wistar rats were submitted to intraperitoneal saline injection (n=10, sham Group), transient middle cerebral artery occlusion (tMCAO) and saline injection (n=10, placebo Group), tMCAO and highly purified fish-oil administration (n=10, T Group). At the two latter groups, twenty-four hours after tMCAO, MRI scans were performed to identify the ischemic regions; the eligible animals were sacrificed, the left parietal bones being removed and subjected to qualitative and quantitative histological and immunohistochemical analysis.
Results: The active hematopoietic surface was maximal at the T-Group, being significantly lower in the P- and S-Groups (p=0.006 and p= 0.017). The MPO positive surface increased significantly in the T-compared to the S-Group (22.57± 0.86 % vs. 18.87± 0.68%, p= 0.004). Arg1 expression was significantly higher (p=0.001), while iNOS expression was lower (p=0.004) in the T- than in the P-Group, but similar to the S-group. The Arg1/iNOS2 ratio was higher in the FO-treated than in the P-group (p<0.001).
Conclusion: the ischemic conditions triggered granulopoiesis and the increase of iNOS2 positive, type M1 macrophage in the rat bone marrow. Fish-oil treatment generated the expansion of overall hematopoietic surface along with normalization of iNOS2, up-regulating the potentially protective Arg1 positive M2 type macrophages and causing a significant shift in the M2/M1 ratio.
Aim: The aim of this study was to investigate the in-vitro efficacy of Glycyrrhetinic acid against Helicobacter pylori (H. pylori) strains, as compared with conventional antibacterial agents.
Methods: A total of 41 H. pylori isolates were used, 6 of which were of standard strains (NCTC 1637), 8 of which were drug-sensitive, and 27 were resistant to drugs isolates. Clarithromycin and metronidazole resistance in all strains of H. pylori were determined by the Epsilometer test (E-test) method. MIC study was performed by using microdilution broth method.
Results: Glycyrrhetinic acid was found to be effective against H. pylori NCTC 1637 in doses of 12.0±4.38 µg/mL, while the MIC value of clinical H. pylori isolates susceptible to antimicrobials was 20.8±10.11 µg/ml. It was found that the MIC values for antimicrobial-sensitive clinical H. pylori isolates was higher when compared with H. pylori NCTC 1637 strains. The MIC values of the standard antimicrobial agents against drug-resistant H. pylori strains were higher than H. pylori NCTC 1637 strains and drug-sensitive H. pylori strains. The MIC value was found to be 14.22±7.77 µg/ml for metronidazole, 3.89±1.90 µg/ml for clarithromycin, 2.33±1.0 µg/ml for amoxicillin, 2.44±0.88 µg/ml for levofloxacin and 4.89±2.47 µg/ml for tetracycline, whereas the MIC value of Glycyrrhetinic acid was 26.67±8.0 µg/ml in metronidazole-resistant H. pylori isolates. Besides, MIC values of the antimicrobials and 18ß-Glycyrrhetinic acid among the strains resistant to clarithromycin were as follows: 3.25±2.12 µg/ml for metronidazole, 9.71±4.54 µg/ml for clarithromycin, 2.06±1.32 µg/ml for amoxicillin, 3.88±4.22 µg/ml for levofloaxacin and 3.25±1.04 µg/mL for tetracycline and 22.0±11.11 µg/ml for Glycyrrhetinic acid.
Conclusion: Glycyrrhetinic acid had significant antimicrobial activity against H. pylori strains. Although further in-vivo studies are needed on antimicrobial activity of Glycyrrhetinic acid, increased resistance to drugs currently used in treatment suggests that Glycyrrhetinic acid may be a potential agent for the treatment of H. pylori.
In recent years, a significant number of epidemiological variations have been observed for fungal infections. In immunocompromised patients, Candida albicans is crucially involved in invasive infections, mostly originating in respiratory tract colonization. The global rise in candidiasis has led researchers to investigate possible correlations between fungal strains virulence profiles and their pathogenic potential, among the most investigated genes being those involved in adherence and biofilm development. In this study, we established the adherence gene profiles of C. albicans strains isolated from respiratory tract secretions in patients hospitalized for cardiovascular diseases and correlated them with the ability of the respective strains to colonize the epithelial cells and form biofilms on the inert substratum. The strains isolated from the lower respiratory tract exhibited the highest adherence capacity and were intensive biofilm producers. The SAP9, ALS3, ALS5, and ALS6 genes were the most frequently detected. There was a significant association between the presence of ALS 3 gene and the cellular substrate colonizing potential of the harboring strains. We also found that the strains expressing SAP9 were more virulent in the phenotypic assays. Detecting the presence of adherence genes from different clinical isolates is a cost-effective tool that would allow researchers to predict the virulence of a certain strain and estimate its potential to adhere to host cells and develop biofilms.
Lung cancer (LC), which includes small-cell lung carcinoma (SCLC) and non-small-cell lung carcinoma (NSCLC), is common and has a high fatality rate. This study aimed to reveal the prognostic mechanisms of LC. GSE30219 was extracted from the Gene Expression Omnibus (GEO) database, and included 293 LC samples and 14 normal lung samples. Differentially expressed genes (DEGs) were identified using the Limma package, and subjected to pathway enrichment analysis using DAVID. MicroRNAs (miRNAs) targeting the DEGs were predicted using Webgestalt. Cytoscape software was used to build a protein-protein interaction (PPI) network and to identify significant network modules. Survival analysis was conducted using Survminer and Survival packages, and validation was performed using The Cancer Genome Atlas (TCGA) dataset. The good and poor prognosis groups contained 518 DEGs. miR-190, miR-493, and miR-218 for the upregulated genes and miR-302, miR-200, and miR-26 for the downregulated genes were predicted. Three network modules (module 1, 2, and 3) were identified from the PPI network. CDK1, MCM10, and NDC80 were the core nodes of module 1, 2, and 3, respectively. In module 1, CDK1 interacted with both CCNB1 and CCNB2. Additionally, CDK1, CCNB1, CCNB2, MCM10, and NDC80 expression levels correlated with clinical survival and were identified as DEGs in both GSE30219 and the TCGA dataset. miR-190, miR-493, miR-218, miR-200, and miR-302 might act in LC by targeting the DEGs. CDK1, CCNB1, CCNB2, MCM10, and NDC80 might also influence the prognosis of LC.
Background: Oxidative damage is of great importance for patients with breast cancer. Thus, studies were performed to identify the relationship between breast cancer and oxidative stress biomarkers.
Objectives: In this study, our aim was to find out the oxidative and antioxidant status, serum thiol-disulphide levels in subjects with breast cancer.
Methods: This study was conducted between March and June 2018 with 82 control subjects (aged between 32-67 years) and 127 breast cancer patients (aged between 27-66 years) (p=0.058) in Selcuk University Faculty of Medicine, Konya, Turkey. Serum myeloperoxidase (MPO), catalase, prolidase were analyzed with kinetic spectrophotometric and thiol-disulphide, ischemia-modified albumin (IMA), ceruloplasmin were detected by colorimetric methods.
Results: Serum levels of catalase [199.3 (16.4-489.9) vs 81.6 (18.2-322.9) (kU/L)], MPO [124±28 vs 101±31 U/L], disulphide [25 (11-61) vs 18 (2-41) µmol/L], IMA [0.66 (0.31-3.30) vs 0.62 (0.19-1.31) absorbance unit (ABSU)] and prolidase levels [2217±538 vs 1456±401 U/L] were higher in patients than control subjects (For all p<0.001 except for IMA p=0.031). Native thiol [342±60 vs 391±52 µmol/L] and total thiol levels [396±56 vs 430±52 µmol/L] were lower in patients compared with the control group (For all p<0.001).
Conclusions: Levels of serum thiol/disulphide and prolidase might be reliable indicators for determining oxidative status in certain patient populations.
Purpose: The implantable bladder cancer (BC) models allow the researchers to perform rapid and useful experiments for BC. We investigated the implantation success of BC cells obtained from Wistar rats (grown in vitro), into bladders of syngeneic Wistar rats, which are commonly used in the laboratories.
Methods: The Nara Bladder Tumor No.2 (NBT-II) BC cells induced with 4-hydroxybutylnitrosamine were grown with passages in Kocaeli University Center for Stem-Cell and Gene-Therapies. After urothelial denudation, 2x106 NBT-II cells were then implanted into bladders of 24 female Wistar rats (aged 7-8 weeks). The rats were randomly divided into four experimental groups; three instillation groups (8 per group) and one sham-operated control group consisting of 6 rats. First, second and third instillation groups were sacrificed at days 7, 14, and 21, respectively, and, bladders were histopathologically evaluated for BC according to WHO / International Society of Urological Pathology.
Results: All tumors were pT1 (including 1 rat that prematurely died at 5th day), except one rat that died prematurely at 8th day had pT2 tumor. Implantation rates were 28.58% (2/7) in the first group, and 42.85% (3/7) in the second, for a cumulative rate of 35.71% (5/14) in these two-groups (until 14th day). Interestingly, there was no tumor in the third group, but there was an inflammatory granulation tissue.
Conclusion: Seeding NBT-II cells into bladders of Wistar rats was described, successfully tested and demonstrated in this study. This implantable BC model of Wistar rats may be improved to increase the success rate of BC cell implantation in new studies with higher number of animals.
Purpose: Ovarian torsion, being a gynecological emergency, requires to be rapidly diagnosed and treated with minimal consequences on ovarian function after the removal of torsion. As ischemia modified albumin (IMA) is considered a good biomarker in diverse ischemic diseases, the aim of our study was to determine the effect of different ovarian torsion/detorsion models on serum and ovarian homogenates levels of IMA in an experimental study.
Methods: IMA was measured in the serum and ovarian homogenates of 7 groups of female rats (10 animals in each group): 1 - control (no intervention); 2 - sham (only laparotomy); 3 - ischemia group: 3 hours ovarian torsion (OT); 4 - 3 hours OT (ischemia), 1 hour simple reperfusion; 5 - 3 hours ischemia, 1 hour controlled reperfusion that was assured during the first two minutes by opening and closing the clips on the ovarian pedicles in 10 seconds intervals, followed by simple reperfusion; 6 - 3 hours ischemia, 24 hours simple reperfusion; 7 - 3 hours OT, 24 hours controlled reperfusion. The results were analyzed by Welch’s ANOVA and Spearman correlation.
Results: Ischemia increases the IMA in both serum and ovarian homogenates compared to control and sham groups. The controlled reperfusion groups had a statistically significant lower IMA in serum compared to simple reperfusion groups. IMA was found to be higher in the ovarian homogenates of simple reperfusion compared to controlled reperfusion groups.
Conclusion: Our results suggest that controlled reperfusion prevent the processes that increase the IMA in ovarian torsion.
Introduction: Experimental acute cerebral ischemia quickly triggers circulating inflammatory cells, provoking infiltration of neutrophils and macrophages in the damaged brain region. N-3 polyunsaturated fatty acids alleviate the ischemic deterioration, however, their potential effect on bone marrow cell mobilization is less known.
Materials and methods: healthy male Wistar rats were submitted to intraperitoneal saline injection (n=10, sham Group), transient middle cerebral artery occlusion (tMCAO) and saline injection (n=10, placebo Group), tMCAO and highly purified fish-oil administration (n=10, T Group). At the two latter groups, twenty-four hours after tMCAO, MRI scans were performed to identify the ischemic regions; the eligible animals were sacrificed, the left parietal bones being removed and subjected to qualitative and quantitative histological and immunohistochemical analysis.
Results: The active hematopoietic surface was maximal at the T-Group, being significantly lower in the P- and S-Groups (p=0.006 and p= 0.017). The MPO positive surface increased significantly in the T-compared to the S-Group (22.57± 0.86 % vs. 18.87± 0.68%, p= 0.004). Arg1 expression was significantly higher (p=0.001), while iNOS expression was lower (p=0.004) in the T- than in the P-Group, but similar to the S-group. The Arg1/iNOS2 ratio was higher in the FO-treated than in the P-group (p<0.001).
Conclusion: the ischemic conditions triggered granulopoiesis and the increase of iNOS2 positive, type M1 macrophage in the rat bone marrow. Fish-oil treatment generated the expansion of overall hematopoietic surface along with normalization of iNOS2, up-regulating the potentially protective Arg1 positive M2 type macrophages and causing a significant shift in the M2/M1 ratio.
Aim: The aim of this study was to investigate the in-vitro efficacy of Glycyrrhetinic acid against Helicobacter pylori (H. pylori) strains, as compared with conventional antibacterial agents.
Methods: A total of 41 H. pylori isolates were used, 6 of which were of standard strains (NCTC 1637), 8 of which were drug-sensitive, and 27 were resistant to drugs isolates. Clarithromycin and metronidazole resistance in all strains of H. pylori were determined by the Epsilometer test (E-test) method. MIC study was performed by using microdilution broth method.
Results: Glycyrrhetinic acid was found to be effective against H. pylori NCTC 1637 in doses of 12.0±4.38 µg/mL, while the MIC value of clinical H. pylori isolates susceptible to antimicrobials was 20.8±10.11 µg/ml. It was found that the MIC values for antimicrobial-sensitive clinical H. pylori isolates was higher when compared with H. pylori NCTC 1637 strains. The MIC values of the standard antimicrobial agents against drug-resistant H. pylori strains were higher than H. pylori NCTC 1637 strains and drug-sensitive H. pylori strains. The MIC value was found to be 14.22±7.77 µg/ml for metronidazole, 3.89±1.90 µg/ml for clarithromycin, 2.33±1.0 µg/ml for amoxicillin, 2.44±0.88 µg/ml for levofloxacin and 4.89±2.47 µg/ml for tetracycline, whereas the MIC value of Glycyrrhetinic acid was 26.67±8.0 µg/ml in metronidazole-resistant H. pylori isolates. Besides, MIC values of the antimicrobials and 18ß-Glycyrrhetinic acid among the strains resistant to clarithromycin were as follows: 3.25±2.12 µg/ml for metronidazole, 9.71±4.54 µg/ml for clarithromycin, 2.06±1.32 µg/ml for amoxicillin, 3.88±4.22 µg/ml for levofloaxacin and 3.25±1.04 µg/mL for tetracycline and 22.0±11.11 µg/ml for Glycyrrhetinic acid.
Conclusion: Glycyrrhetinic acid had significant antimicrobial activity against H. pylori strains. Although further in-vivo studies are needed on antimicrobial activity of Glycyrrhetinic acid, increased resistance to drugs currently used in treatment suggests that Glycyrrhetinic acid may be a potential agent for the treatment of H. pylori.
In recent years, a significant number of epidemiological variations have been observed for fungal infections. In immunocompromised patients, Candida albicans is crucially involved in invasive infections, mostly originating in respiratory tract colonization. The global rise in candidiasis has led researchers to investigate possible correlations between fungal strains virulence profiles and their pathogenic potential, among the most investigated genes being those involved in adherence and biofilm development. In this study, we established the adherence gene profiles of C. albicans strains isolated from respiratory tract secretions in patients hospitalized for cardiovascular diseases and correlated them with the ability of the respective strains to colonize the epithelial cells and form biofilms on the inert substratum. The strains isolated from the lower respiratory tract exhibited the highest adherence capacity and were intensive biofilm producers. The SAP9, ALS3, ALS5, and ALS6 genes were the most frequently detected. There was a significant association between the presence of ALS 3 gene and the cellular substrate colonizing potential of the harboring strains. We also found that the strains expressing SAP9 were more virulent in the phenotypic assays. Detecting the presence of adherence genes from different clinical isolates is a cost-effective tool that would allow researchers to predict the virulence of a certain strain and estimate its potential to adhere to host cells and develop biofilms.