Genomic Analysis of Stress Response against Arsenic in Caenorhabditis elegans

Arsenic, a known human carcinogen, is widely distributed around the world and found in particularly high concentrations in certain regions including Southwestern US, Eastern Europe, India, China, Taiwan and Mexico. Chronic arsenic poisoning affects millions of people worldwide and is associated with increased risk of many diseases including arthrosclerosis, diabetes and cancer. In this study, we explored genome level global responses to high and low levels of arsenic exposure in Caenorhabditis elegans using Affymetrix expression microarrays. This experimental design allows us to do microarray analysis of dose-response relationships of global gene expression patterns. High dose (0.03%) exposure caused stronger global gene expression changes in comparison with low dose (0.003%) exposure, suggesting a positive dose-response correlation. Biological processes such as oxidative stress, and iron metabolism, which were previously reported to be involved in arsenic toxicity studies using cultured cells, experimental animals, and humans, were found to be affected in C. elegans. We performed genome-wide gene expression comparisons between our microarray data and publicly available C. elegans microarray datasets of cadmium, and sediment exposure samples of German rivers Rhine and Elbe. Bioinformatics analysis of arsenic-responsive regulatory networks were done using FastMEDUSA program. FastMEDUSA analysis identified cancer-related genes, particularly genes associated with leukemia, such as dnj-11, which encodes a protein orthologous to the mammalian ZRF1/MIDA1/MPP11/DNAJC2 family of ribosome-associated molecular chaperones. We analyzed the protective functions of several of the identified genes using RNAi. Our study indicates that C. elegans could be a substitute model to study the mechanism of metal toxicity using high-throughput expression data and bioinformatics tools such as FastMEDUSA.     

Reduction in co-trimoxazole resistance inEscherichia coli isolates from urinary tract infections between 1995 and 2005: a multicenter study in Ankara, Turkey

Co-trimoxazole resistance inEscherichia coli isolates from urinary tract infections (UTI) was assessed in 382 strains from 1995 and 510 strains from 2005. The strains were collected from five microbiology laboratories in Ankara, Turkey. Documentation on patient gender, age and outpatient/inpatient status was collected in 2005, but not in 1995. The resistance percentages were 751% in 1995 and 55.5% in 2005. This reduction in resistance percentage was statistically significant, overall in all except two of the participating laboratories. The resistance percentage in 2005 was 61.1% for children (n=208) and 51.2% for adults (n=258), 53.7% for females (n=380) and 60.8% for males (n=130), and 55.3% for outpatients (n=400) and 56.4% for inpatients (n=110). The reduction in resistance is believed to be a consequence of reduced usage. Although decreased, the level of co-trimoxazole resistance remains high, and continued avoidance of its use for empiric treatment of UTI in Turkey appears to be an appropriate strategy.     

Loss of GM130 in breast cancer cells and its effects on cell migration,invasion and polarity

Spatially distinct pools of the small GTPase Cdc42 were observed, but the major focus of research so far has been to investigate its signaling at the plasma membrane. We recently showed that the Golgi pool of Cdc42 is relevant for cell polarity and that it is regulated by GM130, a Golgi matrix protein. Loss of GM130 abrogated cell polarity and consistent with the notion that polarity is frequently impaired in cancer, we found that GM130 is downregulated in colorectal cancer. Whether the loss of GM130 solely affects polarity, or whether it affects other processes relevant for tumorigenesis remains unclear. In a panel of breast cancer cells lines, we investigated the consequences of GM130 depletion on traits of relevance for tumor progression, such as survival, proliferation, adhesion, migration and invasion. We show that cellular assays that depend on polarity, such as chemotaxis and wound scratch assays, are only of limited use to investigate the role of polarity modulators in cancer. Depletion of GM130 increases cellular velocity and increases the invasiveness of breast cancer cells, therefore supporting the view that alterations of polarity contribute to tumor progression.     

Training Top-Down Attention Improves Performance on a Triple-Conjunction Search Task

Training has been shown to improve perceptual performance on limited sets of stimuli. However, whether training can generally improve top-down biasing of visual search in a target-nonspecific manner remains unknown. We trained subjects over ten days on a visual search task, challenging them with a novel target (top-down goal) on every trial, while bottom-up uncertainty (distribution of distractors) remained constant. We analyzed the changes in saccade statistics and visual behavior over the course of training by recording eye movements as subjects performed the task. Subjects became experts at this task, with twofold increased performance, decreased fixation duration, and stronger tendency to guide gaze toward items with color and spatial frequency (but not necessarily orientation) that resembled the target, suggesting improved general top-down biasing of search.     

Comparison of oxidative stress and antioxidant capacity before and after running exercises in both sexes

Background: It has been difficult to determine, from the published literature, whether men or women have higher levels of exercise-induced oxidative stress.Objective: The aim of this study was to compare variations between the sexes in lipid hydroperoxide (LPO), superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and lactate dehydrogenase (LDH) after 3 different running exercises performed at the same speed.Methods: Eligible participants were healthy university students of both sexes. The participants performed running exercise tests at distances of 800, 1500, and 3000 m at a speed of 10 km/h. Blood samples were taken from the participants just before and immediately after the running activities to determine LPO, SOD, CAT, GR, and LDH, and these measures were compared both before and after exercise and between the sexes.Results: A total of 17 young and healthy, but not physically trained, students (n = 8 men; mean age, 22.00 years; n = 9 women; mean age, 21.78 years) participated in this study. Height, weight, and maximum oxygen consumption values were significantly higher in men than in women (P = 0.01). Significant gender effects were found in LPO levels at 3000 m (F = 5.51; P = 0.03) and in SOD activity at 800 m (F = 7.92; P = 0.01) and 3000 m (F = 6.05; P = 0.03). CAT activity also differed between the sexes at 800 m (F = 15.67; P = 0.01) and 1500 m (F = 6.55; P = 0.02). However, no significant gender-time interaction effect was observed for any measurement at the 800-, 1500-, and 3000-m distances.Conclusions: Changes in LPO, SOD, and CAT activities at different running distances were not different between men and women over time because of a nonsignificant gender-time interaction. With regard to changes in oxidative stress, men and women had similar responses to exercise at the same absolute workload, despite significant differences in physical characteristics.     

Involvement of cAMP-response element binding protein-1 in arachidonic acid-induced vascular smooth muscle cell motility

In addition to their role in many vital cellular functions, arachidonic acid (AA) and its eicosanoid metabolites are involved in the pathogenesis of several diseases, including atherosclerosis and cancer. To understand the potential mechanisms by which these lipid molecules could influence the disease processes, particularly cardiovascular diseases, we studied AA's effects on vascular smooth muscle cell (VSMC) motility and the role of cAMP-response element binding protein-1 (CREB-1) in this process. AA exerted differential effects on VSMC motility; at lower doses, it stimulated motility, whereas at higher doses, it was inhibitory. AA-induced VSMC motility requires its conversion via the lipoxygenase (LOX) and cyclooxygenase (COX) pathways. AA stimulated the phosphorylation of extracellular signal-regulated kinases (ERKs), Jun N-terminal kinases (JNKs), and p38 mitogen-activated protein kinase (p38MAPK) in a time-dependent manner, and blockade of these serine/threonine kinases significantly attenuated AA-induced VSMC motility. In addition, AA stimulated CREB-1 phosphorylation and activity in a manner that was also dependent on its metabolic conversion via the LOX and COX pathways and the activation of ERKs and p38MAPK but not JNKs. Furthermore, suppression of CREB-1 activation inhibited AA-induced VSMC motility. 15(S)-Hydroxyeicosatetraenoic acid and prostaglandin F2alpha, the 15-LOX and COX metabolites of AA, respectively, that are produced by VSMC at lower doses, were also found to stimulate motility in these cells. Together, these results suggest that AA induces VSMC motility by complex mechanisms involving its metabolism via the LOX and COX pathways as well as the ERK- and p38MAPK-dependent and JNK-independent activation of CREB-1.     

Oxidative cleavage of DNA by homo- and heteronuclear Cu(II)-Mn(II) complexes of an oxime-type ligand

Novel homodinuclear Cu(II) (K1), heterodinuclear Cu(II)-Mn(II) (K2) and homotrinuclear Cu(II) (K3) complexes with a novel oxime-type ligand have been prepared and their nucleolytic activities on pCYTEXP were established by neutral agarose gel electrophoresis. The analyses of the cleavage products obtained electrophoretically indicate that although the examined complexes induces very similar conformational changes on supercoiled DNA by converting supercoiled form to nicked form than linear form in a sequential manner as the complex concentration or reaction period is increased, K3 is less effective than the two others. The oxime complexes were nucleolytically active at physiological pH values but the activities of K1 or K2 were diminished by increasing the pH of the reaction mixture. In contrast, K3 makes dominantly single strand nicking by producing nicked circles on DNA at almost all the applied pH values. Metal complex induced DNA cleavage was also tested for inhibition by various radical scavengers as superoxide dismutase (SOD), azide, thiourea and potassium iodide. The antioxidants inhibited the nucleolytic acitivities of the oxime complexes but SOD afforded no protection indicating that the nucleolytic mechanism involves of copper and/or manganese complex-mediated reactive oxygen species such as hydroxyl radicals being responsible for the oxidative DNA cleavage.