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Kinds of Time Points Selected AbstractsAttachment transfer among Swedish and German adolescents: A prospective longitudinal studyPERSONAL RELATIONSHIPS, Issue 3 2006WOLFGANG FRIEDLMEIER This prospective longitudinal study investigated whether and in what way different functions of attachment relationships are transferred from parents to reciprocal relationship partners in adolescence. Furthermore, the impact of nationality, romantic relationship status, and individual differences in perceived attachment history and current attachment orientation on the timing and extent of transfer was examined. Adolescents from Sweden and Germany were studied over a 12- to 15-month time span. As predicted, the transfer generally unfolded in a step-by-step process in cross-sectional analyses. However, the predicted direction of transfer from parents to peers could not be confirmed in the prospective analyses. Adolescents who had formed a romantic relationship between assessments showed a stronger transfer from parents to peers compared to those who had not. German adolescents had transferred to a larger extent at Time Point 1, but Swedish adolescents caught up by Time Point 2. Finally, the combination of an insecure history with mother and high current anxiety was linked to a particularly high degree of prospective attachment transfer, whereas an insecure history with mother combined with high current avoidance predicted a particularly low degree of prospective transfer. [source] Automatic Extraction of ECG Strips from Continuous 12-lead Holter Recordings for QT Analysis at Prescheduled versus Optimized Time PointsANNALS OF NONINVASIVE ELECTROCARDIOLOGY, Issue 2009Fabio Badilini Ph.D. Background: Continuous 12-lead ECG monitoring (Holter) in early-phase pharmaceutical studies is today widely used as an ideal platform to extract discrete ECGs for analysis. The extraction process is typically performed manually by trained readers using commercial Holter processing systems. Methods: Antares, a novel method for automatic 12-lead extraction from continuous Holter recordings applying minimal noise criteria and heart-rate stability conditions is presented. A set of 12-lead Holter recordings from healthy subjects administered with sotalol is used to compare ECG extractions at fixed time points with ECG extractions generated by Antares optimizing noise and heart rate inside 5 minute windows centered around each expected time point of interest. Results: Global, low- and high-frequency noise content of extracted ECGs was significantly reduced via optimized approach by Antares. Heart rate was also slightly reduced (from 69 ± 13 to 64 ± 13 bpm, P < 0.05). Similarly, the corrected QT interval from optimized extractions was significantly reduced (QTcB from 414 ± 32 to 402 ± 30 ms, P < 0.05). Using only baseline data, and after adjusting for intersubject variability, the standard deviation (SD) of QT intervals was highly reduced with optimized extraction (SD of QTcF from 11 ± 8 to 7 ± 2 ms, P < 0.05). Conclusions: Extraction of discrete 12-lead ECG strips from continuous Holter generates less noisy and more stable ECGs leading to more robust QTc data, thereby potentially facilitating the assessment of ECG effects on clinical trials. [source] Intra-amniotic endotoxin accelerates lung maturation in fetal rabbitsACTA PAEDIATRICA, Issue 1 2001Kristina Bry The hypothesis that endotoxin in amniotic fluid accelerates fetal lung maturation was tested. On day 25 of gestation, LPS (5 ,g/fetus) was injected intra-amniotically into one uterine horn of eight New Zealand white rabbits, whereas the contralateral amniotic sacs were injected with saline vehicle. The fetuses were delivered 48 h after LPS administration and their lungs were studied. One dam went into premature labor prior to the 48 h time point and was excluded from the study. Mean white cell counts in amniotic fluid and bronchoalveolar lavage fluid from LPS-treated fetuses were increased 3.2-fold (p= 0.04) and 9.9-fold (p= 0.04), respectively. Fetal weights and lung weights were not affected by LPS. Surfactant protein SP-A and SP-B mRNA expressions in LPS-treated fetuses were increased 2.3-fold (p= 0.03) and 1.4-fold (p= 0.04), respectively. Static lung compliance was increased in animals treated with LPS (p= 0.001). Lungs from LPS-treated animals had better aeration than those of controls. Mean volume of inflation-fixed lungs of LPS-treated fetuses was 1.7 times greater than that of controls (p= 0.03). Conclusion: Intra-uterine exposure to LPS increases surfactant protein expression and improves lung stability and aeration in preterm animals. [source] Rapid and easy semi-quantitative evaluation method for diacylglycerol and inositol-1,4,5-trisphosphate generation in orexin receptor signallingACTA PHYSIOLOGICA, Issue 3 2010M. E. Ekholm Abstract Aim:, Fluorescent protein-based indicators have enabled measurement of intracellular signals previously nearly inaccessible for studies. However, indicators showing intracellular translocation upon response suffer from serious limitations, especially the very time-consuming data collection. We therefore set out in this study to evaluate whether fixing and counting cells showing translocation could mend this issue. Methods:, Altogether three different genetically encoded indicators for diacylglycerol and inositol-1,4,5-trisphosphate were transiently expressed in Chinese hamster ovary cells stably expressing human OX1 orexin receptors. Upon stimulation with orexin-A, the cells were fixed with six different protocols. Results:, Different protocols showed clear differences in their ability to preserve the indicator's localization (i.e. translocation after stimulus) and its fluorescence, and the best results for each indicator were obtained with a different protocol. The concentration,response data obtained with cell counting are mostly comparable to the real-time translocation and biochemical data. Conclusion:, The counting method, as used here, works at single time point and looses the single-cell-quantitative aspect. However, it also has some useful properties. First, it easily allows processing of a 100- to 1000-fold higher cell numbers than real-time imaging producing statistically consistent population-quantitative data much faster. Secondly, it does not require expensive real-time imaging equipment. Fluorescence in fixed cells can also be quantitated, though this analysis would be more time-consuming than cell counting. Thirdly, in addition to the quantitative data collection, the method could be applied for identifying responsive cells. This might be very useful in identification of e.g. orexin-responding neurones in a large population of non-responsive cells in primary cultures. [source] Atomoxetine treatment in adults with attention-deficit/hyperactivity disorder and comorbid social anxiety disorderDEPRESSION AND ANXIETY, Issue 3 2009Lenard A. Adler M.D. Abstract Background: To evaluate the effect of atomoxetine (ATX) on attention-deficit/hyperactivity disorder (ADHD) and comorbid social anxiety disorder in adults. Methods: Randomized, double-blind, placebo-controlled, conducted in adults with ADHD and social anxiety disorder. Patients received 40,100,mg ATX (n=224) or placebo (n=218) for 14 weeks following a 2-week placebo lead-in period. Efficacy measures included the Conners' Adult ADHD Rating Scale: Investigator-Rated: Screening Version (CAARS:Inv:SV), Liebowitz Social Anxiety Scale (LSAS), Clinical Global Impression-Overall-Severity (CGI-O-S), State-Trait Anxiety Inventory (STAI), Social Adjustment Scale-Self Report (SAS), and Adult ADHD Quality of Life Scale-29 (AAQoL). Safety and tolerability were also assessed. Results: ATX mean change (,8.7±10.0) from baseline (29.6±10.4) on CAARS:Inv:SV Total ADHD Symptoms score was significantly greater than placebo mean change (,5.6±10.2) from baseline (31.2±9.4; P<.001). ATX mean change (,22.9±25.3) from baseline (85.3±23.6) on LSAS Total score was significant compared to placebo mean change (,14.4±20.3) from baseline (82.1±21.3; P<.001). The visit-wise analysis revealed greater improvement on the CAARS:Inv:SV Total ADHD Symptoms score and LSAS Total score for ATX at every time point throughout the study (P values ,.012). Mean changes in CGI-O-S, STAI-Trait Anxiety scores, and AAQoL Total score were significantly greater for ATX compared to placebo. Mean change for both groups on STAI-State Anxiety scores was comparable. Improvement on SAS for ATX compared to placebo was not significant. Rates of insomnia, nausea, dry mouth, and dizziness were higher with ATX than with placebo. Discontinuation rates due to treatment-emergent adverse events were similar between groups. Conclusions: ATX monotherapy effectively improved symptoms of ADHD and comorbid social anxiety disorder in adults and was well tolerated. Depression and Anxiety, 2009. Published 2009 Wiley-Liss, Inc. [source] A Randomized Trial to Determine the Influence of Laser Therapy, Monopolar Radiofrequency Treatment, and Intense Pulsed Light Therapy Administered Immediately after Hyaluronic Acid Gel ImplantationDERMATOLOGIC SURGERY, Issue 5 2007MITCHEL P. GOLDMAN MD BACKGROUND Hyaluronic acid-based dermal fillers, such as hyaluronic acid gel (Restylane, Q-Medical AB, Uppsala, Sweden), are widely used for tissue augmentation of the nasolabial folds. Additional dermatologic treatments using infrared light, radiofrequency (RF), and intense pulsed light (IPL) are also important tools for facial rejuvenation. This study was designed to evaluate whether these therapies could be safely administered immediately after hyaluronic acid gel treatment without compromising the effect of the dermal filler. OBJECTIVE The objective of this study was to confirm or refute any possible subtractive effects of augmentation of the nasolabial folds when followed by 1,320-nm Nd:YAG laser, 1,450-nm diode laser, monopolar RF, and/or IPL treatments. METHODS Thirty-six patients with prominent nasolabial folds were treated with hyaluronic acid gel implantation on one side of the face and hyaluronic acid gel followed by one of the nonablative laser/RF/IPL therapies on the contralateral side of the face. RESULTS There were no statistically significant differences between wrinkle severity or global aesthetic scores for hyaluronic acid gel implantation alone and hyaluronic acid gel with laser/RF/IPL treatment at any time point. In a small sample, histologic changes were not apparent after laser/RF/IPL treatment. CONCLUSIONS Based on this small pilot study, laser, RF, and IPL treatments can safely be administered immediately after hyaluronic acid gel implantation without reduction in overall clinical effect. [source] Four-Year Follow-up on Endovascular Radiofrequency Obliteration of Great Saphenous RefluxDERMATOLOGIC SURGERY, Issue 2 2005Robert F. Merchant MD Background Endovascular radiofrequency obliteration has been used since 1998 as an alternative to conventional vein stripping surgery for elimination of saphenous vein insufficiency. Objective To demonstrate the long-term efficacy of this treatment modality. Methods Data were prospectively collected in a multicenter ongoing registry. Only great saphenous vein above-knee treatments were included in this study. Eight hundred ninety patients (1,078 limbs) were treated prior to November 2003 at 32 centers. Clinical and duplex ultrasound follow-up was performed at 1 week, 6 months, and 1, 2, 3, and 4 years. Results Among 1,078 limbs treated, 858 were available for follow-up within 1 week, 446 at 6 months, 384 at 1 year, 210 at 2 years, 114 at 3 years, and 98 at 4 years. The vein occlusion rates were 91.0%, 88.8%, 86.2%, 84.2%, and 88.8%, respectively; the reflux-free rates were 91.0%, 89.3%, 86.2%, 86.0%, and 85.7%, respectively; and the varicose vein recurrence rates were 7.2%, 13.5%, 17.1%, 14.0%, and 21.4%, respectively, at each follow-up time point at 6 months, and 1, 2, 3, and 4 years. Patient symptom improvement persisted over 4 years. Conclusions Endovascular temperature-controlled radiofrequency obliteration of saphenous vein reflux exhibits an enduring treatment efficacy clinically, anatomically, and hemodynamically up to 4 years following treatment. ROBERT F. MERCHANT, MD, AND OLIVIER PICHOT, MD, ARE PAID CONSULTANTS TO VNUS MEDICAL TECHNOLOGIES, WHICH PROVIDED FINANCIAL SUPPORT FOR THIS STUDY. [source] Effectiveness of Microporous Polysaccharide Hemospheres for Achieving Hemostasis in Mohs Micrographic SurgeryDERMATOLOGIC SURGERY, Issue 6 2004FRCPC, Stephen R. Tan MD Background. Microporous polysaccharide hemospheres consist of controlled-porosity spherical particles manufactured from bioinert plant polysaccharide. Microporous polysaccharide hemospheres facilitate hemostasis by rapidly absorbing the fluid component of blood, concentrating platelets and clotting factors to accelerate blood clotting. Objective. The objective was to compare a microporous polysaccharide hemosphere bandage and electrocautery in achieving hemostasis. Methods. Twenty-four patients with a total of 48 stages of Mohs micrographic surgery were included. Patients were stratified by whether or not they were taking anticoagulant medications. Within each group, patients were randomized to receive either the microporous polysaccharide hemosphere bandage or electrocautery. Outcomes included bleeding through the dressing (early time point) and active bleeding upon dressing removal (late time point). Results. Nineteen patients not taking anticoagulants had 40 stages, of which 18 received the study bandage and 22 received electrocautery. The remaining 5 patients on anticoagulants had 8 stages, of which 4 received the study bandage and 4 received electrocautery. In both total and subgroup analysis, there was a higher incidence of bleeding through the dressing with the study bandage (p<0.05), but no increase in the incidence of active bleeding upon dressing removal (p>0.05). Conclusion. The microporous polysaccharide hemosphere study bandage had an increased incidence of bleeding through the dressing compared to electrocautery, but did not have an increased incidence of active bleeding upon dressing removal. [source] Impaired lactation in mice expressing dominant-negative FADD in mammary epitheliumDEVELOPMENTAL DYNAMICS, Issue 4 2009Mark Shackleton Abstract The Fas-associated death domain (FADD/Mort1) adaptor protein was originally identified as a key mediator of apoptosis, although pleiotropic functions for FADD have also been reported. FADD-mediated tumoricidal effects have been described in breast cancer cells; however, its physiological role in normal mammary gland epithelium is not well understood. To determine the role of FADD signaling during mammary gland development, we generated transgenic mice overexpressing dominant-negative FADD (DN-FADD) in mammary epithelium, using the steroid responsive mouse mammary tumor virus promoter. Transgenic mice exhibited a perturbation in lactation resulting in impaired milk production and pup growth retardation. Reduced expansion of alveoli was evident during early lactation with extensive shedding of luminal alveolar cells. Significantly more TUNEL (terminal deoxynucleotidyl transferase,mediated deoxyuridinetriphosphate nick end-labeling)-positive cells were present at this time point and a subsequent increase in bromodeoxyuridine-positive cells was observed. These findings suggest a role for FADD in maintaining the survival of mammary secretory alveolar cells after the establishment of lactation. Developmental Dynamics 238:1010,1016, 2009. © 2009 Wiley-Liss, Inc. [source] Maternal hypoxia increases the activity of MMPs and decreases the expression of TIMPs in the brain of neonatal ratsDEVELOPMENTAL NEUROBIOLOGY, Issue 3 2010Wenni Tong Abstract A recent study has shown that increased activity of matrix metalloproteinases-2 and metalloproteinases-9 (MMP-2 and MMP-9) has detrimental effect on the brain after neonatal hypoxia. The present study determined the effect of maternal hypoxia on neuronal survivability and the activity of MMP-2 and MMP-9, as well as the expression of tissue inhibitors of metalloproteinase 1 and 2 (TIMP-1 and TIMP-2) in the brain of neonatal rats. Pregnant rats were exposed to 10.5% oxygen for 6 days from the gestation day 15 to day 21. Pups were sacrificed at day 0, 4, 7, 14, and 21 after birth. Body weight and brain weight of the pups were measured at each time point. The activity of MMP-2 and MMP-9 and the protein abundance of TIMP-1 and TIMP-2 were determined by zymography and Western blotting, respectively. The tissue distribution of MMPs was examined by immunofluorescence staining. The neuronal death was detected by Nissl staining. Maternal hypoxia caused significant decreases in body and brain size, increased activity of MMP-2 at day 0, and increased MMP-9 at day 0 and 4. The increased activity of the MMPs was accompanied by an overall tendency towards a reduced expression of TIMPs at all ages with the significance observed for TIMPs at day 0, 4, and 7. Immunofluorescence analysis showed an increased expression of MMP-2, MMP-9 in the hippocampus at day 0 and 4. Nissl staining revealed significant cell death in the hippocampus at day 0, 4, and 7. Functional tests showed worse neurobehavioral outcomes in the hypoxic animals. © 2009 Wiley Periodicals, Inc. Develop Neurobiol 2010 [source] Time course analysis of gene expression during light-induced photoreceptor cell death and regeneration in albino zebrafishDEVELOPMENTAL NEUROBIOLOGY, Issue 8 2007Sean C. Kassen Abstract Constant intense light causes apoptosis of rod and cone photoreceptors in adult albino zebrafish. The photoreceptors subsequently regenerate from proliferating inner nuclear layer (INL) progenitor cells that migrate to the outer nuclear layer (ONL) and differentiate into rods and cones. To identify gene expression changes during this photoreceptor regeneration response, a microarray analysis was performed at five time points during the light treatment. The time course included an early time point during photoreceptor death (16 h), later time points during progenitor cell proliferation and migration (31, 51, and 68 h) and a 96 h time point, which likely corresponds to the initial photoreceptor differentiation. Mean expression values for each gene were calculated at each time point relative to the control (0 h light exposure) and statistical analysis by one-way ANOVA identified 4567 genes exhibiting significant changes in gene expression along the time course. The genes within this data set were clustered based on their temporal expression patterns and proposed functions. Quantitative real-time PCR validated the microarray expression profiles for selected genes, including stat3 whose expression increased markedly during the light exposure. Based on immunoblots, both total and activated Stat3 protein expression also increased during the light treatment. Immunolocalization of Stat3 on retinal tissue sections demonstrated increased expression in photoreceptors and Müller glia by 16 h of light exposure. Some of the Stat3-positive Müller cells expressed PCNA at 31 h, suggesting that Stat3 may play a role in signaling a subset of Müller cells to proliferate during the regeneration response. © 2007 Wiley Periodicals, Inc. Develop Neurobiol, 2007. [source] Comparison of the effects of early handling and early deprivation on maternal care in the ratDEVELOPMENTAL PSYCHOBIOLOGY, Issue 4 2001Christopher R. Pryce Abstract It has been reported in the rat that postnatal manipulations can induce robust and persistent effects on offspring neurobiology and behavior, mediated in part via effects on maternal care. There have, however, been few studies of the effects of postnatal manipulations on maternal care. Here, we describe and compare the effects on maternal behavior on postnatal days 1,12 of two manipulations, early handling (EH, 15-min isolation per day) and early deprivation (ED, 4-hr isolation per day), relative to our normal postnatal husbandry procedure. Maternal behavior was measured at five time points across the dark phase of the reversed L:D cycle. EH yielded an increase in arched-back nursing across several time points but did not affect any other behavior. ED stimulated a bout of maternal behavior such that licking and arched-back nursing were increased at the time of dam,litter reunion, although not at any other time point. Neither EH nor ED affected weaning weight significantly. Importantly, within-treatment variation was high relative to these between-treatment effects. © 2001 John Wiley & Sons, Inc. Dev Psychobiol 38: 239,251, 2001 [source] Serum concentrations of high-mobility group box chromosomal protein 1 before and after exposure to the surgical stress of thoracic esophagectomy: a predictor of clinical course after surgery?DISEASES OF THE ESOPHAGUS, Issue 1 2006K. Suda SUMMARY., High-mobility group box chromosomal protein 1 (HMGB-1) has recently been shown as an important late mediator of endotoxin shock, intra-abdominal sepsis, and acute lung injury. However, its role in the systemic inflammatory response syndrome after major surgical stress, which may lead to multiple organ dysfunction syndrome, has not been thoroughly investigated. We hypothesized that serum HMGB-1 participates in the pathogenesis of postoperative organ system dysfunction after exposure to major surgical stress. A prospective clinical study was performed to consecutive patients (n = 24) with carcinoma of the thoracic esophagus who underwent transthoracic esophagectomy with three field lymph node resection between 1998 and 2003 at Keio University Hospital, Japan. Serum HMGB-1 concentrations were measured by enzyme-linked immunosorbent assay. Preoperative serum HMGB-1 levels correlated with postoperative duration of SIRS, mechanical ventilation, and intensive care unit stay. Three of the 24 patients had serious postoperative complications: sepsis in two, and acute lung injury in one. Serum HMGB-1 levels in patients without complications increased within the first 24 h postoperatively, remained high during postoperative days 2,3, and then decreased gradually by postoperative day 7. In patients with serious complications, serum HMGB-1 was significantly higher than that found in patients without postoperative complications at every time point except postoperative day 2. Preoperative serum HMGB-1 concentration seems to be an important predictor of the postoperative clinical course. Transthoracic esophagectomy induces an increase in HMGB-1 in serum even in patients without complications. Postoperative serum HMGB-1 concentrations were higher in patients who developed complications, and may be a predictive marker for complications in this setting. [source] Aortic Valve Closure: Relation to Tissue Velocities by Doppler and Speckle Tracking in Patients with Infarction and at High Heart RatesECHOCARDIOGRAPHY, Issue 4 2010Ph.D., Svein A. Aase M.Sc. Aim: To resolve the event in tissue Doppler (TDI)- and speckle tracking-based velocity/time curves that most accurately represent aortic valve closure (AVC) in infarcted ventricles and at high heart rates. Methods: We studied the timing of AVC in 13 patients with myocardial infarction and in 8 patients at peak dobutamine stress echo. An acquisition setup for recording alternating B-mode and TDI image frames was used to achieve the same frame rate in both cases (mean 136.7 frames per second [FPS] for infarcted ventricles, mean 136.9 FPS for high heart rates). The reference method was visual assessment of AVC in the high frame rate narrow sector B-mode images of the aortic valve. Results: The initial negative velocities after ejection in the velocity/time curves occurred before AVC, 44.9 ± 21.0 msec before the reference in the high heart rate material, and 25.2 ± 15.2 msec before the reference in the infarction material. Using this time point as a marker for AVC may cause inaccuracies when estimating end-systolic strain. A more accurate but still a practical marker for AVC was the time point of zero crossing after the initial negative velocities after ejection, 5.4 ± 15.3 msec before the reference in high heart rates and 8.2 ± 12.9 msec after the reference in the infarction material. Conclusion: The suggested marker of AVC at high heart rate and in infarcted ventricles was the time point of zero crossing after the initial negative velocities after ejection in velocity/time curves. (Echocardiography 2010;27:363-369) [source] Electrospun Silk Fibroin Mats for Tissue EngineeringENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 3 2008A. Alessandrino Abstract Processing Silk Fibroin (SF) with electrospinning (ES) offers a very attractive opportunity for producing a variety of 2D and 3D matrices with great potential for tissue regeneration and repair due to the superior biocompatibility and mechanical properties of SF. Different combinations of ES parameters were explored to investigate the best experimental set-up related to the dimension and uniformity of the fibers in the electrospun silk fibroin (ES-SF) mats. Using SEM it was found that the ES-SF mats contain uniform fibers with a diameter in the nanometric range obtained by electrospinning a 7.5,% w/v SF solution in formic acid, with an electric field of 2.4,kV/cm and a spinneret-collector distance of 10,cm. FT-IR and DSC analyses were performed to investigate the structure of the ES-SF mats before and after immersion in methanol for different times (5, 10, and 15,min). The methanol treatment was able to promote the crystallization of SF by conformational transition of random coil and other poorly ordered conformations (turns and bends) to the ,-sheet structure. The degree of crystallinity was enhanced as shown by the trend of both the FT-IR crystallinity index and the melting/decomposition peak temperature (from DSC). To study the cytocompatibility of ES-SF mats, tests with L929 murine fibroblasts were carried out. Samples were seeded with the cells and incubated for 1, 3, and 7,days at 37,°C. At each time point, SEM investigations and Alamar blue tests were performed. The SEM images showed cell adhesion and proliferation just after 1,day and cell confluence at 7,days. Alamar blue test demonstrated that there were very low differences between cell viability on ES-SF mats and the tissue culture plastic control. [source] Radioprotective effects of Daflon against genotoxicity induced by gamma irradiation in human cultured lymphocytesENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 9 2009Seyed Jalal Hosseinimehr Abstract The ability of Daflon to protect against genotoxicity induced by gamma irradiation has been investigated in vivo and in vitro in cultured lymphocytes from healthy human volunteers. Peripheral human blood samples were collected predose (10 min before) and 1, 2, and 3 hr after a single oral ingestion of 1000 mg of Daflon. At each time point, whole blood was exposed in vitro to 150 cGy of cobalt-60 gamma rays, and then the lymphocytes were cultured with mitogenic stimulation to determine the micronuclei in cytokinesis-blocked binucleated cells. For each volunteer, the results showed a significant increase in the incidence of micronuclei after exposure to gamma irradiation as compared to control unexposed samples. As early as 1 hr after Daflon administration, a significant decrease in the incidence of micronuclei was observed in comparison with similarly irradiated lymphocytes collected before administration. The maximum protection was reached 1 hr after administration of Daflon with a significant decrease in the frequency of micronuclei of 40%. These findings suggest the possible application of Daflon for the protection of human lymphocytes from the genetic damage and side effects induced by gamma irradiation. Environ. Mol. Mutagen. 2009. © 2009 Wiley-Liss, Inc. [source] Alterations in gene expression profiles and the DNA-damage response in ionizing radiation-exposed TK6 cells,,ENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 2-3 2005Gregory S. Akerman Abstract Identifying genes that are differentially expressed in response to DNA damage may help elucidate markers for genetic damage and provide insight into the cellular responses to specific genotoxic agents. We utilized cDNA microarrays to develop gene expression profiles for ionizing radiation-exposed human lymphoblastoid TK6 cells. In order to relate changes in the expression profiles to biological responses, the effects of ionizing radiation on cell viability, cloning efficiency, and micronucleus formation were measured. TK6 cells were exposed to 0.5, 1, 5, 10, and 20 Gy ionizing radiation and cultured for 4 or 24 hr. A significant (P < 0.0001) decrease in cloning efficiency was observed at all doses at 4 and 24 hr after exposure. Flow cytometry revealed significant decreases in cell viability at 24 hr in cells exposed to 5 (P < 0.001), 10 (P < 0.0001), and 20 Gy (P < 0.0001). An increase in micronucleus frequency occurred at both 4 and 24 hr at 0.5 and 1 Gy; however, insufficient binucleated cells were present for analysis at the higher doses. Gene expression profiles were developed from mRNA isolated from cells exposed to 5, 10, and 20 Gy using a 350 gene human cDNA array platform. Overall, more genes were differentially expressed at 24-hr than at the 4-hr time point. The genes upregulated (> 1.5-fold) or downregulated (< 0.67-fold) at 4 hr were those primarily involved in the cessation of the cell cycle, cellular detoxification pathways, DNA repair, and apoptosis. At 24 hr, glutathione-associated genes were induced in addition to genes involved in apoptosis. Genes involved in cell cycle progression and mitosis were downregulated at 24 hr. Real-time quantitative PCR was used to confirm the microarray results and to evaluate expression levels of selected genes at the low doses (0.5 and 1.0 Gy). The expression profiles reflect the cellular and molecular responses to ionizing radiation related to the recognition of DNA damage, a halt in progression through the cell cycle, activation of DNA-repair pathways, and the promotion of apoptosis. Environ. Mol. Mutagen., 2005. Published 2005 Wiley-Liss, Inc. [source] Influence of hydroxypropyl-,-cyclodextrin on the extraction and biodegradation of phenanthrene in soilENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2004Brian J. Reid Abstract A study was conducted to investigate the effect of hydroxypropyl-,-cyclodextrin (HPCD) on the aging and biodegradation of phenanthrene (PHE) in soil. Soil was spiked with PHE at 25 mgPHE/kgSOIL and HPCD at a range of concentrations from 0 to 3.5 gHPCD/kgSOIL and aged for 1, 84, and 322 d. At each time point, a variety of analyses were performed to assess the loss and aging of the PHE in the soil. Methods included determination of total PHE remaining, dichloromethane (DCM) and butan-1-ol (BuOH) extractions, and determination of PHE extractable by an aqueous HPCD shake extraction. Mineralization assays were also carried out to assess the availability of the PHE to a PHE-degrading bacterial inoculum. It was found that the presence of HPCD in the soils increased PHE loss from the aged soil systems, particularly at the higher application rates. Dichloromethane and BuOH extractabilities were reduced with aging and increasing HPCD concentration, as was the amount of PHE that was extractable using an aqueous HPCD shake extraction or that was available for mineralization. The DCM and BuOH extraction yielded similar results, and both greatly overestimated the availability of the PHE to the degraders, whereas the HPCD extraction results were very similar to that of PHE biodegradation. This study indicates that cyclodextrins have potential for use as alternatives to surfactants in enhancing the desorption/solubilization and degradation of recalcitrant organic contaminants in soil. [source] Cardiac regeneration by progenitor cells , bedside before bench?EUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 7 2005J. Bauersachs Abstract Recent experimental and clinical trials give rise to the hope that progenitor cells could replace scar tissue after myocardial infarction with healthy functional myocardium. However, while a significant increase in left ventricular ejection fraction has been described after progenitor cell transplantation in several clinical trials, long-term results are lacking, and the mechanisms underlying the improvement of ejection fraction are unclear. Therefore, the efficacy of progenitor cell transplantation after myocardial infarction has not been established, and potential problems may have been underestimated. In-depth laboratory and animal studies are needed to determine the best cell type, optimal amount of cells, and time point for transplantation. Treatment of patients with progenitor cells outside well controlled prospective trials should be avoided. [source] The effect of a community dental service outreach programme on the confidence of undergraduate students to treat children: a pilot studyEUROPEAN JOURNAL OF DENTAL EDUCATION, Issue 1 2007M. Lindsay Hunter Objective:, To examine the effect of a community dental service (CDS) outreach teaching programme on undergraduates' confidence to undertake a range of paediatric dental procedures. Method:, Eighteen final year dental students completed a questionnaire prior to, and following participation in an outreach teaching programme. At each time point, the students were asked to identify how confident they felt to carry out a range of procedures commonly encountered in the treatment of children, employing a Likert scale modified to comprise six points where a rating of 1 represented ,not at all confident' and a rating of 6 ,very confident'. Results:, The distribution of scores at each time point indicated that students were more confident to carry out each of the listed procedures following participation in the outreach teaching programme than they had been on completion of their paediatric dentistry sessions within the School of Dentistry. At the individual student level, 16 of the 18 students indicated that they were, overall, more confident following their placement than previously. Conclusions:, It can be concluded that the long-established CDS outreach teaching programme run by the School of Dentistry, Wales College of Medicine in conjunction with the staff of Cardiff and Vale NHS Trust is a valuable adjunct to undergraduate teaching in paediatric dentistry. [source] Heart rate variability in beta-thalassemia patientsEUROPEAN JOURNAL OF HAEMATOLOGY, Issue 5 2009Wasarut Rutjanaprom Abstract Background:, Cardiac failure remains the major cause of death in beta-thalassemia major (TM). Reduced heart rate variability (HRV) is associated with a higher risk of arrhythmias after myocardial infarction and heart failure. We evaluated HRV in TM patients and its relationship with hemodynamics and echocardiographic parameters during a 6-month follow-up. Methods:, Thirty-four TM patients (19 ± 10 yr) and 20 healthy subjects (17 ± 6 yr) were evaluated. Hematologic, biochemical, echocardiographic and HRV parameters were determined at entry and at 6-month follow-up. Time and frequency domain HRV parameters were analyzed from 24-h recorded electrocardiograms. All TM patients received blood transfusion and chelation therapy. Results:, Both time and frequency domain HRV parameters were markedly reduced in TM patients, compared to the control. The significantly improved HRV was seen in correlation with higher hemoglobin (Hb) level when compared within TM group at different time point. No correlation was seen between HRV and serum ferritin, reactive oxygen species (ROS) and non-transferrin bound iron (NTBI). Conclusion:, HRV is depressed in TM patients. HRV was significantly correlated with Hb level, suggesting that anemia greatly influences the cardiac autonomic balance. [source] CLINICAL STUDY/BIOMARKER: Phosphatidylethanol: normalization during detoxification, gender aspects and correlation with other biomarkers and self-reportsADDICTION BIOLOGY, Issue 1 2010Friedrich Martin Wurst ABSTRACT Phosphatidylethanol (PEth) is a direct ethanol metabolite, and has recently attracted attention as biomarker of ethanol intake. The aims of the current study are: (1) to characterize the normalization time of PEth in larger samples than previously conducted; (2) to elucidate potential gender differences; and (3) to report the correlation of PEth with other biomarkers and self-reported alcohol consumption. Fifty-seven alcohol-dependent patients (ICD 10 F 10.25; 9 females, 48 males) entering medical detoxification at three study sites were enrolled. The study sample was comprised of 48 males and 9 females, with mean age 43.5. Mean gamma glutamyl transpeptidase (GGT) was 209.61 U/l, average mean corpuscular volume (MCV) was 97.35 fl, mean carbohydrate deficient transferrin (%CDT) was 8.68, and mean total ethanol intake in the last 7 days was 1653 g. PEth was measured in heparinized whole blood with a high-pressure liquid chromatography method, while GGT, MCV and %CDT were measured using routine methods. PEth levels at day 1 of detoxification ranged between 0.63 and 26.95 µmol/l (6.22 mean, 4.70 median, SD 4.97). There were no false negatives at day 1. Sensitivities for the other biomarkers were 40.4% for MCV, 73.1% for GGT and 69.2% for %CDT, respectively. No gender differences were found for PEth levels at any time point. Our data suggest that PEth is (1) a suitable intermediate term marker of ethanol intake in both sexes; and (2) sensitivity is extraordinary high in alcohol dependent patients. The results add further evidence to the data that suggest that PEth has potential as a candidate for a sensitive and specific biomarker, which reflects longer-lasting intake of higher amounts of alcohol and seemingly has the above mentioned certain advantages over traditional biomarkers. [source] Cocaine increases medial prefrontal cortical glutamate overflow in cocaine-sensitized rats: a time course studyEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 6 2004Jason M. Williams Abstract Excitatory amino acid transmission within mesocorticolimbic brain pathways is thought to play an important role in behavioural sensitization to psychomotor stimulants. The current studies evaluated a time course of the effects of cocaine on extracellular glutamate levels within the medial prefrontal cortex (mPFC) following increasing periods of withdrawal from repeated cocaine exposure. Male Sprague,Dawley rats underwent stereotaxic surgeries and were pretreated daily with saline (1 mL/kg/day × 4 days, i.p.) or cocaine (15 mg/kg/day × 4 days, i.p.) and withdrawn for 1, 7 or 30 days. After withdrawal rats were challenged with the same dose of saline or cocaine and in vivo microdialysis of the mPFC was conducted with concurrent analysis of locomotor activity. Animals that were withdrawn from repeated daily cocaine for 1 day and 7 days displayed an augmentation in cocaine-induced mPFC glutamate levels compared to saline and acute control subjects, which were similarly unaffected by cocaine challenge. At the 7 day time point, a subset of animals that received repeated cocaine did not express behavioural sensitization, nor did these animals exhibit the enhancement in mPFC glutamate in response to cocaine challenge. In contrast to these early effects, 30 days of withdrawal resulted in no significant changes in cocaine-induced mPFC glutamate levels regardless of the pretreatment or behavioural response. These data suggest that repeated cocaine administration transiently increases cocaine-induced glutamate levels in the mPFC during the first week of withdrawal, which may play an important role in the development of behavioural sensitization to cocaine. [source] Opposing actions of neuropeptide Y and light on the expression of circadian clock genes in the mouse suprachiasmatic nucleiEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 1 2002Elizabeth S. Maywood Abstract The circadian clockwork of the hypothalamic suprachiasmatic nuclei (SCN) is synchronized by light and by nonphotic cues. The core timing mechanism is cell-autonomous, based on an autoregulatory transcriptional/translational feedback loop of circadian genes and their products. This study investigated the effects of neuropeptide Y (NPY), a potent nonphotic resetting cue, and its interaction with light in regulating clock gene expression in the SCN in vivo. Injection of NPY adjacent to the SCN and transfer to darkness 7 h before scheduled lights out, shifted the circadian activity,rest cycle. Exposure to light for 1 h immediately after NPY infusion blocked this behavioural response. NPY-induced shifts were accompanied by suppression of both mPer1 and mPer2 mRNA in the SCN, assessed 3 h after infusion. mPer mRNAs were not altered 1 h after infusion. Levels of mClock mRNA or mCLOCK immunoreactivity in the SCN were not affected by NPY at either time point. In parallel to the behavioural response, the NPY-induced suppression of mPer genes in the SCN was attenuated when a light pulse was delivered immediately after the infusion. These results identify mPer1 and mPer2 as molecular targets for both photic and nonphotic (NPY-induced) resetting of the clockwork, and support a synthetic model of circadian entrainment based upon convergent up- and downregulation of mPer expression. [source] Taurine selectively modulates the secretory activity of vasopressin neurons in conscious ratsEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 7 2001Mario Engelmann Abstract Previous experiments have shown that a 10-min forced swimming session triggers the release of vasopressin from somata and dendrites, but not axon terminals, of neurons of the hypothalamic,neurohypophysial system. To further investigate regulatory mechanisms underlying this dissociated release, we forced male Wistar rats to swim in warm (20 °C) water and monitored release of the potentially inhibitory amino acids gamma amino butyric acid (GABA) and taurine into the hypothalamic supraoptic nucleus using microdialysis. Forced swimming caused a significant increase in the release of taurine (up to 350%; P < 0.05 vs. prestress release), but not GABA. To reveal the physiological significance of centrally released taurine, the specific taurine antagonist 6-aminomethyl-3-methyl-4H-1,2,4-benzothiadiazine-1,1-dioxide was administered into the supraoptic nucleus via retrodialysis. Administration of this antagonist caused a significant increase in the release of vasopressin within the supraoptic nucleus and into the blood both under basal conditions and during stress (up to 800%; P < 0.05 vs. basal values), without affecting hypothalamic or plasma oxytocin. Local administration of the GABAA receptor antagonist bicuculline, in contrast, failed to influence vasopressin secretion at either time point. In a separate series of in vivo electrophysiological experiments, administration of the same dosage of the taurine antagonist into the supraoptic nucleus via microdialysis resulted in an increased electrical activity of identified vasopressinergic, but not oxytocinergic, neurons. Taken together our data demonstrate that taurine is released within the supraoptic nucleus during physical/emotional stress. Furthermore, at the level of the supraoptic nucleus, taurine inhibits not only the electrical activity of vasopressin neurons but also acts as an inhibitor of both central and peripheral vasopressin secretion during different physiological states. [source] Regulation of hippocampal cell adhesion molecules NCAM and L1 by contextual fear conditioning is dependent upon time and stressor intensityEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 9 2000J. Joaquķn Merino Abstract Cell adhesion molecules (CAMs) of the immunoglobulin superfamily, NCAM and L1, as well as the post-translational addition of ,-2,8-linked polysialic acid (PSA) homopolymers to NCAM (PSA,NCAM), have been implicated in the neural mechanisms underlying memory formation. Given that the degree of stress elicited by the training situation is one of the key factors that influence consolidation processes, this study questioned whether training rats under different stressor intensities (0.2, 0.4, or 1 mA shock intensity) in a contextual fear conditioning task might regulate subsequent expression of NCAM, PSA,NCAM and L1 in the hippocampus, as evaluated immediately after testing rats for conditioning at 12 and 24 h after training. Behavioural inhibition (evaluated as a ,freezing' index) at testing and post-testing plasma corticosterone levels were also assessed. The results showed that 12 h post-training, conditioned animals displayed reduced NCAM, but increased L1, expression. At this time point, the group trained at the highest shock intensity (1 mA) also presented decreased PSA,NCAM expression. Analyses performed 24 h post-training indicated that the 1 mA group exhibited increased NCAM and L1 expression, but decreased expression of PSA,NCAM levels. In addition, L1 values that presented a shock intensity-dependent U-shaped pattern were also increased in the group trained at the lowest shock condition (0.2 mA) and remained unchanged in the intermediate shock condition (0.4 mA). Freezing and corticosterone values at both testing times were positively related with shock intensity experienced at training. Therefore, our results show a complex regulation of CAMs of the immunoglobulin superfamily in the hippocampus that depends upon stressor intensity and time factors. In addition, the pattern of CAMs expression found in the 1 mA group (which is the one that shows higher post-training corticosterone levels and develops the stronger and longer-lasting levels of fear conditioning) supports the view that, after a first phase of synaptic de-adherence during consolidation, NCAM and L1 might participate in the stabilization of selected synapses underlying the establishment of long-term memory for contextual fear conditioning, and suggests that glucocorticoids might play a role in the observed regulation of CAMs. [source] Abundant genetic variation in transcript level during early Drosophila developmentEVOLUTION AND DEVELOPMENT, Issue 6 2008Sergey V. Nuzhdin SUMMARY Variation in gene expression may underlie many important evolutionary traits. However, it is not known at what stage in organismal development changes in gene expression are most likely to result in changes in phenotype. One widely held belief is that changes in early development are more likely to result in changes in downstream phenotypes. In order to discover how much genetic variation for transcript level is present in natural populations, we studied zygotic gene expression in nine inbred lines of Drosophila melanogaster at two time points in their development. We find abundant variation for transcript level both between lines and over time; close to half of all expressed genes show a significant line effect at either time point. We examine the contribution of maternally loaded genes to this variation, as well as the contribution of variation in upstream genes to variation in their downstream targets in two well-studied gene regulatory networks. Finally, we estimate the dimensionality of gene expression in these two networks and find that,despite large numbers of varying genes,there appear to be only two factors controlling this variation. [source] Delayed onset of midline netrin expression in Artemia franciscana coincides with commissural axon growth and provides evidence for homology of midline cells in distantly related arthropodsEVOLUTION AND DEVELOPMENT, Issue 2 2007Molly Duman-Scheel SUMMARY Although many similarities in arthropod central nervous systems (CNS) development exist, differences in midline cell formation and ventral nerve cord axonogenesis have been noted in arthropods. It is possible that changes in the expression of axon guidance molecules such as Netrin, which functions during commissural axon guidance in Drosophila and many other organisms, may parallel these differences. In this investigation, we analyze this hypothesis by examining Netrin accumulation during development of the brine shrimp Artemia franciscana, a branchiopod crustacean. An Artemia franciscana netrin (afrnet) orthologue was cloned. An antibody to the afrNet protein was generated and used to examine the pattern of afrNet accumulation during Artemia development. Despite differences between Drosophila and Artemia nerve cord development, examination of afrNet accumulation suggests that this protein functions to regulate commissure formation during Artemia CNS development. However, detection of afrNet at the midline and on commissural axons occurs at a relatively later time point in Artemia as compared with Drosophila. Detection of afrNet in a subset of midline cells that closely resemble Netrin-expressing cells at the Drosophila midline provides evidence for homology of midline cells in arthropods. Expression of Netrins in many other tissues is comparable, suggesting that Netrin proteins may play many conserved roles during arthropod development. [source] Defining the caspase-containing apoptotic machinery contributing to cornification in human epidermal equivalentsEXPERIMENTAL DERMATOLOGY, Issue 1 2006Vijaya Chaturvedi Abstract:, Whether terminal differentiation/stratum corneum formation of keratinocytes (KCs) represents a form of programmed cell death, utilizing mediators of classical apoptosis, is unclear. Apoptosis, an evolutionarily conserved death process, is comprised of extrinsic and intrinsic pathways, which converge using caspase 3. To define upstream and downstream caspases involved in terminal differentiation, we utilized human epidermal equivalents (EEs). Using submerged cultures comprised of human KCs, EEs were sequentially analyzed before and after being raised to an air/liquid (A/L) interface at 3,24 h intervals. At each time point, EEs were analyzed morphologically and for specific enzyme activity to distinguish different initiator (caspases 1, 2, 8, 9) and effector caspases (3, 6, 7). Terminal differentiation began at 6,8 h, as defined by stratum corneum with loricirin expression and completed at 18,24 h producing an epidermis resembling normal skin. Enzyme activity for caspases 1, 2, 3, 6, 7, 8, and 9 (but not 4, 5) was enhanced (>two-fold nmol/mg/h) at 3,6 h compared with submerged cultures. Processing of caspase 14 occurred at 18 h, and cleaved caspase 14 was increased at 24 h. Activated caspase 3-positive and terminal deoxynucleotidyl transferase-mediated nick end labeling-positive KCs were identified in EEs at 3,6 h corresponding to initiation sites of terminal differentiation. Addition of caspase inhibitors reduced levels of involucrin and loricrin in EEs raised to an A/L interface. We conclude caspases function as important death effectors strategically positioned at intersection of intrinsic and extrinsic pathways in KCs undergoing stratum corneum formation. [source] Expression of melanoma-associated antigens in melanoma cell culturesEXPERIMENTAL DERMATOLOGY, Issue 7 2005Mirjana Urosevic Abstract:, The efficiency of melanoma immunotherapy appears to depend on both melanoma- and immune system-specific factors. Melanoma-specific factors include melanoma-associated antigen (MAA) expression as well as HLA class I molecule expression. We investigated the expression of five MAA , Melan-A/MART-1, tyrosinase, gp100, MAGE-1 and MAGE-3 , by means of FACS analysis in 50 melanoma cell cultures and compared them to the cultures of human foreskin-derived melanocytes and melanoma cell line UKRV-Mel2. Melan-A, tyrosinase and gp100 expression was frequently reduced in melanoma cell cultures, compared to that in foreskin melanocytes, whereas MAGE-1 and MAGE-3 expression showed variable degree of upregulation, compared to that in foreskin melanocytes. The expression of all tested MAA demonstrated high interindividual variability. We further show that cell cultures derived from the same tissue sample are oligoclonal in nature, by demonstrating the presence of up to three cell populations bearing distinct MAA profile. Analysing samples derived from the same patient but each at a different time point, we show that MAA expression profile changes over time either in positive (increase) or in negative (decrease) direction. Finally, we demonstrate that brain metastasis-derived cell cultures significantly overexpress Melan-A and MAGE-3, compared to primary tumours and other metastatic sites (P -value range: 0.05,0.001). Elucidation of the MAA expression patterns and the kinetics within the same patient as well as during the course of the disease may help improve current and develop new immunotherapeutic strategies. [source] | ||||||||||||||||||||||||||||||||||||||||