bluuebunny/crossref_metadata_2025 huggingface dataset:
Within the Limits of Deliberative Reason Alone
Authors: Thomassen Lasse
Published: March 2007

In this article, I take Habermas's treatment of civil disobedience as a litmus test of the way in which Habermas relates to the imperfectness of democracy. The case of civil disobedience, which Habermas deems to be a normal part of a mature constitutional democracy, shows that Habermas is ultimately unable to submit all decisions and distinctions to the public use of reason as envisaged in his deliberative account of democracy. As a consequence, I argue that we must take the imperfectness of democracy as constitutive and our conceptualization of civil disobedience and democracy must start from there. The reading of Habermas and the argument for an alternative approach are inspired by the work of Jacques Derrida, central to which is an idea of democracy and justice ‘to-come’ focusing our attention on the performative role of civil disobedience in creating more legitimate laws.
Internationalising Media Studies
Authors: Tomaselli Keyan , Teer-Tomaselli Ruth E.
Published: March 2007

A discussion of the history of journalism and mass communication studies, in relation to its alter ego, media studies, provides the backdrop to an analysis of the role played by the South African Communication Association (SACOMM) during the 1980s and after apartheid, up to 2006. The central argument is that SACOMM was criss-crossed by a particular kind of internal family argument in which specific contradictions were being negotiated: (1) pro-apartheid vs anti-apartheid (during the 1980s); (2) realists vs idealists; and (3) communication science vs media studies. The way these antagonisms had been negotiated by 2006 provides some insight into the way that media studies assisted in internationalizing the association and in exposing communication science to other paradigms.
A novel proteoliposomal vaccine induces antitumor immunity against follicular lymphoma
Authors: Neelapu Sattva S. , Gause Barry L. , Harvey Linda , Lee Seung-Tae , Frye Andrea Robin , Horton Jessie , Robb Richard J. , Popescu Mircea C. , Kwak Larry W.
Published: March 2007

Clinical studies suggest that treatment with vaccines comprised of idiotype protein may be associated with improved clinical outcome in follicular lymphoma patients. The time-consuming process required to generate patient-specific vaccines is a major limitation, however. Here we report results of a pilot clinical trial with a novel autologous, tumor-derived proteoliposome vaccine formulation that could be rapidly produced within a single day. Vaccination was safe, induced autologous tumor-specific type 1 cytokine responses in 5 out of 10 follicular lymphoma patients, and was associated with induction of a sustained complete response in one patient. Other patients had large tumor burdens and progressed after a median duration of 8 months. These results suggest that further testing of this vaccine formulation, particularly in the setting of minimal disease, is warranted. Furthermore, the proteoliposome formulation may provide a model for vaccine development for other human cancers, for which tumor-associated antigens need not be defined.
Proteomic patterns predict acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation
Authors: Weissinger Eva M. , Schiffer Eric , Hertenstein Bernd , Ferrara James L. , Holler Ernst , Stadler Michael , Kolb Hans-Jochem , Zander Axel , Zürbig Petra , Kellmann Markus , Ganser Arnold
Published: March 2007

Acute graft-versus-host disease (aGvHD) contributes significantly to morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Diagnosis of GvHD is mainly based on clinical features and tissue biopsies. A noninvasive, unbiased laboratory test for GvHD diagnosis does not exist. Here we describe the application of capillary electrophoresis coupled online with mass spectrometry (CE-MS) to 13 samples from 10 patients with aGvHD of grade II or more and 50 control samples from 23 patients without GvHD. About 170 GvHD-specific polypeptides were detected and a tentatively aGvHD-specific model consisting of 31 polypeptides was chosen, allowing correct classification of 13 of 13 (sensitivity 100.0% [95% confidence interval {CI} 75.1 to 100.0]) aGvHD samples and 49 of 50 (specificity 98.0% [95% CI 89.3 to 99.7]) control samples of the training set. The subsequent blinded evaluation of 599 samples enabled diagnosis of aGvHD greater than grade II, even prior to clinical diagnosis, with a sensitivity of 83.1% (95% CI 73.1 to 87.9) and a specificity of 75.6% (95% CI 71.6 to 79.4). Thus, high-resolution proteome analysis represents an unbiased laboratory-based screening method, enabling diagnosis, and possibly enabling preemptive therapy.
Friend of GATA-1–independent transcriptional repression: a novel mode of GATA-1 function
Authors: Johnson Kirby D. , Boyer Meghan E. , Kang Jeong-Ah , Wickrema Amittha , Cantor Alan B. , Bresnick Emery H.
Published: March 2007

The GATA-1–interacting protein Friend Of GATA-1 (FOG-1) is essential for the proper transcriptional activation and repression of numerous GATA-1 target genes. Although FOG-1–independent activation by GATA-1 has been described, all known examples of GATA-1–mediated repression are FOG-1 dependent. In the GATA-1–null G1E cell line, estrogen receptor ligand binding domain (ER) chimeras of either wild-type GATA-1 or a FOG-1–binding defective mutant of GATA-1 repressed several genes similarly upon activation with β-estradiol. Repression also occurred in a FOG-1–null cell line expressing ER–GATA-1 and during ex vivo erythropoiesis. At the Lyl1 and Rgs18 loci, we found highly restricted occupancy by GATA-1 and GATA-2, indicating that these genes are direct targets of GATA factor regulation. The identification of genes repressed by GATA-1 independent of FOG-1 defines a novel mode of GATA-1–mediated transcriptional regulation.
Dominant inheritance of retinal ganglion cell resistance to optic nerve crush in mice
Authors: Li Yan , Semaan Sheila J , Schlamp Cassandra L , Nickells Robert W
Published: March 2007

Background

Several neurodegenerative diseases are influenced by complex genetics that affect an individual's susceptibility, disease severity, and rate of progression. One such disease is glaucoma, a chronic neurodegenerative condition of the eye that targets and stimulates apoptosis of CNS neurons called retinal ganglion cells. Since ganglion cell death is intrinsic, it is reasonable that the genes that control this process may contribute to the complex genetics that affect ganglion cell susceptibility to disease. To determine if genetic background influences susceptibility to optic nerve damage, leading to ganglion cell death, we performed optic nerve crush on 15 different inbred lines of mice and measured ganglion cell loss. Resistant and susceptible strains were used in a reciprocal breeding strategy to examine the inheritance pattern of the resistance phenotype. Because earlier studies had implicated Bax as a susceptibility allele for ganglion cell death in the chronic neurodegenerative disease glaucoma, we conducted allelic segregation analysis and mRNA quantification to assess this gene as a candidate for the cell death phenotype.

Results

Inbred lines showed varying levels of susceptibility to optic nerve crush. DBA/2J mice were most resistant and BALB/cByJ mice were most susceptible. F1 mice from these lines inherited the DBA/2J phenotype, while N2 backcross mice exhibited the BALB/cByJ phenotype. F2 mice exhibited an intermediate phenotype. A Wright Formula calculation suggested as few as 2 dominant loci were linked to the resistance phenotype, which was corroborated by a Punnett Square analysis of the distribution of the mean phenotype in each cross. The levels of latent Bax mRNA were the same in both lines, and Bax alleles did not segregate with phenotype in N2 and F2 mice.

Conclusion

Inbred mice show different levels of resistance to optic nerve crush. The resistance phenotype is heritable in a dominant fashion involving relatively few loci. Bax was excluded as a candidate gene for this phenotype.
Estradiol valerate and alcohol intake: dose-response assessments
Authors: Quirarte Gina L , Reid Larry D , de la Teja I Sofía Ledesma , Reid Meta L , Sánchez Marco A , Díaz-Trujillo Arnulfo , Aguilar-Vazquez Azucena , Prado-Alcalá Roberto A
Published: March 2007

Background

An injection of estradiol valerate (EV) provides estradiol for a prolonged period. Recent research indicates that a single 2.0 mg injection of EV modifies a female rat's appetite for alcoholic beverages. This research extends the initial research by assessing 8 doses of EV (from .001 to 2.0 mg/female rat), as well assessing the effects of 2.0 mg EV in females with ovariectomies.

Results

With the administration of EV, there was a dose-related loss of bodyweight reaching the maximum loss, when it occurred, at about 4 days after injections. Subsequently, rats returned to gaining weight regularly. Of the doses tested, only the 2.0 mg dose produced a consistent increase in intake of ethanol during the time previous research indicated that the rats would show enhanced intakes. There was, however, a dose-related trend for smaller doses to enhance intakes. Rats with ovariectomies showed a similar pattern of effects, to intact rats, with the 2 mg dose. After extensive histories of intake of alcohol, both placebo and EV-treated females had estradiol levels below the average measured in females without a history of alcohol-intake.

Conclusion

The data support the conclusion that pharmacological doses of estradiol can produce enduring changes that are manifest as an enhanced appetite for alcoholic beverages. The effect can occur among females without ovaries.
Response of lymphocyte subsets and cytokines to Shenyang prescription in Sprague-Dawley rats with tongue squamous cell carcinomas induced by 4NQO
Authors: Jiang Canhua , Ye Dongxia , Qiu Weiliu , Zhang Xiuli , Zhang Zhiyuan , He Di , Zhang Ping , Chen Wantao
Published: March 2007

Background

The study was designed to investigate immunocompetence in relation to cancer progression in rat and to assess the effect of the traditional Chinese anti-cancer medicine, "Shenyang" prescription, on immunity.

Methods

4-Nitroquinoline-1-oxide (4NQO) was administered to 80 Sprague-Dawley (SD) rats via the drinking water for up to 36 weeks. Tongue squamous cell carcinoma (SCC) was confirmed by pathological examination in 61 rats. "Shenyang" prescription was administered to subgroups of these rats, and blood samples were taken before and after treatment. Lymphocyte subsets were determined by flow cytometry. Serum Th1 and Th2-type cytokines were assessed by an enzyme-linked immunosorbent assay.

Results

As the cancer progressed at the tongue root, the percentage of CD3+CD4+ T lymphocytes and NK cells and the levels of IFN-γ and IL-2 decreased gradually, while the percentage of CD3+CD8+ T lymphocytes and the levels of IL-4 and IL-10 increased. The CD4+/CD8+ ratios were lower in the cancer groups than in the control group. However, after administering "Shenyang" prescription, the levels of CD3+CD4+ T lymphocytes, NK cells, IFN-γ and IL-2 increased, while the CD3+CD8+ T lymphocyte counts and the levels of IL-4 and IL-10 decreased.

Conclusion

4NQO-induced lesions were good models for exploring oral cavity carcinogenesis. The rats with 4NQO-induced SCC demonstrated abnormalities in lymphocyte subsets and a shift from Th1-type to Th2-type, which were good models for assessing the effect of anticancer agent on immunity. Oral cancer progression was associated with an aggressive disturbance of immune function. "Shenyang" prescription has the ability to improve the disturbance of immune function.
Oxidative Upregulation of Bcl‐2 in Healthy Lymphocytes
Authors: CRISTOFANON SILVIA , NUCCITELLI SILVIA , D'ALESSIO MARIA , RADOGNA FLAVIA , DE NICOLA MILENA , BERGAMASCHI ANTONIO , CERELLA CLAUDIA , MAGRINI ANDREA , DIEDERICH MARC , GHIBELLI LINA
Published: March 2007

Abstract:  In many cell systems, pharmacological glutathione (GSH) depletion with the GSH neosynthesis inhibitor buthionine sulfoximine (BSO) leads to cell death and highly sensitizes tumor cells to apoptosis induced by standard chemotherapeutic agents. However, some tumor cells upregulate Bcl‐2 in response to BSO, thus surviving the treatment and failing to be chemosensitized. Cell lines of monocytic and lymphocytic origins respond to BSO treatment in an opposite way, lymphocytes being chemosensitized and unable to transactivate Bcl‐2. In this article we investigate the response to BSO of lymphocytes freshly isolated from peripheral blood of healthy donors. After ensuring that standard separation procedures do not alter per se lymphocytes redox equilibrium nor Bcl‐2 levels in the first 24 h of culture, we show that BSO treatment promotes the upregulation of Bcl‐2, with a mechanism involving the increased radical production consequent to GSH depletion. Thus, BSO treatment may increase the differential cytocidal effect of cytotoxic drugs in tumor versus normal lymphocytes.
Intracellular Pro‐oxidant Activity of Melatonin Deprives U937 Cells of Reduced Glutathione without Affecting Glutathione Peroxidase Activity
Authors: ALBERTINI MARIA CRISTINA , RADOGNA FLAVIA , ACCORSI AUGUSTO , UGUCCIONI FRANCESCO , PATERNOSTER LAURA , CERELLA CLAUDIA , DE NICOLA MILENA , D'ALESSIO MARIA , BERGAMASCHI ANTONIO , MAGRINI ANDREA , GHIBELLI LINA
Published: March 2007

Abstract:  It was long believed that melatonin might counteract intracellular oxidative stress because it was shown to potentiate antioxidant endogenous defences, and to increase the activity of many antioxidant enzymes. However, it is now becoming evident that when radicals are measured within cells, melatonin increases, rather than decreasing, radical production. Herein we demonstrate a pro‐oxidant effect of melatonin in U937 cells by showing an increase of intracellular oxidative species and a depletion of glutathione (GSH). The activity of glutathione peroxidase is not modified by melatonin treatment as it does occur in other experimental models.
Role of Reactive Oxygen Species in Kv Channel Inhibition and Vasoconstriction Induced by TP Receptor Activation in Rat Pulmonary Arteries
Authors: COGOLLUDO ANGEL , FRAZZIANO GIOVANNA , COBEÑO LAURA , MORENO LAURA , LODI FEDERICA , VILLAMOR EDUARDO , TAMARGO JUAN , PEREZ‐VIZCAINO FRANCISCO
Published: March 2007

Abstract:  Voltage‐gated potassium channels (Kv) and thromboxane A2 (TXA2) have been involved in several forms of human and experimental pulmonary hypertension. We have reported that the TXA2 analog U46619, via activation of TP receptors and PKCζ, inhibited Kv currents in rat pulmonary artery smooth muscle cells (PASMC), increased cytosolic calcium, and induced a contractile response in isolated rat and piglet pulmonary arteries (PA). Herein, we have analyzed the role of reactive oxygen species (ROS) in this signaling pathway. In rat PA, U46619 increased dichlorofluorescein fluorescence, an indicator of intracellular hydrogen peroxide, and this effect was prevented by the NADPH oxidase inhibitor apocynin and by polyethyleneglycol‐catalase (PEG‐catalase, a membrane‐permeable form of catalase). U46619 inhibited Kv currents in native PASMC and these effects were strongly inhibited by apocynin. The contractile responses to U46619 in isolated PA were inhibited by PEG‐catalase and the NADPH oxidase inhibitors diphenylene iodonium (DPI) and apocynin. A membrane permeable of hydrogen peroxide, t‐butyl hydroperoxide, also inhibited Kv currents and induced a contractile response. Activation of NADPH oxidase and the subsequent production of hydrogen peroxide are involved in the Kv channel inhibition and the contractile response induced by TP receptor activation in rat PA.
Ataxia‐Telangiectasia‐Mutated‐Dependent Activation of Ku in Human Fibroblasts Exposed to Hydrogen Peroxide
Authors: LEE JONG HWA , KIM KYUNG HWAN , MORIO TOMOHIRO , KIM HYEYOUNG
Published: March 2007

Abstract:  DNA is damaged in cells during cell replication, by infection, or by various environmental stresses. The damaged cells stop cell cycle, repair damaged DNA, and when repaired progress into the next cell cycle stage. But when the attempt to repair the damage fails, the cells undergo apoptosis. The most deleterious damage of all is double‐strand DNA breaks (DSBs), where ATM (ataxia‐telangiectasia‐mutated) serves as a sensor. The ATM pathway culminates in DNA repair through nonhomologous end‐joining or through homologous recombination. Upon DNA damage, the DNA repair protein Ku70/80 translocates into the nucleus, which may be mediated by ATM. Previously, we found that pancreatic acinar cells undergo apoptosis upon oxidative stress, and the cell death stems from nuclear loss of Ku70/80. This study aims to investigate whether ATM has a role in Ku activation and prevention of cell death induced by oxidative stress (hydrogen peroxide) using A‐T fibroblasts stably transfected with human full‐length ATM cDNA or empty vector. As a result, hydrogen peroxide‐induced cell death was augmented in A‐T cells transfected with empty vector while cell death was prevented in A‐T fibroblasts stably transfected with human full‐length ATM cDNA. Ku DNA‐binding activity induced by hydrogen peroxide treatment was increased in the A‐T fibroblasts stably transfected with human full‐length ATM cDNA compared to that in A‐T cells transfected with empty vector. The results suggest that ATM may be essential for Ku activation to repair DNA damage from oxidative stress and prevent cell death caused by oxidative stress.
Regulation of 2‐Deoxy‐d‐Glucose Transport, Lactate Metabolism, and MMP‐2 Secretion by the Hypoxia Mimetic Cobalt Chloride in Articular Chondrocytes
Authors: MOBASHERI ALI , PLATT NICOLA , THORPE COLIN , SHAKIBAEI MEHDI
Published: March 2007

Abstract:  Articular cartilage is an avascular tissue with significantly reduced levels of oxygen and nutrients compared to plasma and synovial fluid. Therefore, chondrocyte survival and cartilage homeostasis require effective mechanisms for oxygen and nutrient signaling. To gain a better understanding of the mechanisms responsible for oxygen and nutrient sensing in chondrocytes, we investigated the effects of hypoxic stimulation induced by cobalt chloride treatment (a hypoxia‐mimetic) on glucose uptake and lactate production in chondrocytes. We also studied the effects of cobalt chloride and glucose deprivation on the expression and secretion of active MMP‐2. Primary cultures of articular chondrocytes were either maintained in 20% O2 (normoxia) or exposed to the hypoxia‐mimetic cobalt chloride for up to 24 h at the following concentrations: 15 μM, 37.5 μM, and 75 μM. Glucose transport was determined by measuring the net uptake of nonmetabolizable 2‐deoxy‐d‐[2, 6‐3H] glucose into chondrocytes. Active MMP‐2 secretion was assayed by gelatin zymography. Lactic acid production was assayed using a lactate kit. Exposure to cobalt chloride significantly increased the uptake of 2‐deoxy‐d‐[2, 6‐3H] glucose and the production of lactate. Glucose deprivation and cobalt chloride treatment increased levels of active MMP‐2 in the culture medium. Our results suggest that these metabolic alterations are important events during adaptation to hypoxia. Upregulation of MMP‐2 and the build‐up of lactic acid will have detrimental effects on the extracellular matrix and may contribute to the pathogenesis and progression of osteoarthritis (OA).
Determination of Oxidative Stress Status and Concentration of TGF‐β1 in the Blood and Saliva of Osteoporotic Subjects
Authors: YOUSEFZADEH GHOLAMREZA , LARIJANI BAGHER , MOHAMMADIRAD AZADEH , HESHMAT RAMIN , DEHGHAN GHOLAMREZA , RAHIMI ROJA , ABDOLLAHI MOHAMMAD
Published: March 2007

Abstract:  Preliminary reports indicate the influence of oxidative stress and interleukins, particularly TGF‐β1, in maintenance of bone mass. This study was designed to determine any possible variations of cellular lipid peroxidation, the total antioxidant power, and concentration of TGF‐β1 in blood and saliva of osteoporotic subjects in comparison to healthy people. Blood and saliva samples of 22 osteoporotic women and 22 age‐matched healthy women were collected. Samples were analyzed for thiobarbituric acid‐reactive substances (TBARS) as a marker of lipid peroxidation, ferric reducing ability (total antioxidant power, TAP), and concentration of TGF‐β1. The blood and saliva TAP (mean ± SD) of osteoporotic subjects was significantly lower than that of healthy controls (606.65 ± 119.13 vs. 665.64 ± 63.73 mmol/L and 560.43 ± 84.70 vs. 612.05 ± 81.5, respectively). Blood and saliva TBARS (mean ± SD) of osteoporotic subjects were significantly higher than those of healthy controls (0.30 ± 0.04 vs. 0.26 ± 0.04 and 0.23 ± 0.03 vs. 0.16 ± 0.04 μmol/L, respectively). Concentrations of TGF‐β1 (mean ± SD) in plasma and saliva of osteoporotic subjects were not different in comparison to healthy subjects. Results indicate that persons with osteoporosis have an increased oxidative stress that is not accompanied by changes in TGF‐β1 levels. Use of supplementary antioxidants in osteoporotic patients may be helpful.
Gene Expression Modulation in A549 Human Lung Cells in Response to Combustion‐Generated Nano‐Sized Particles
Authors: ARENZ ANDREA , HELLWEG CHRISTINE E. , STOJICIC NEVENA , BAUMSTARK‐KHAN CHRISTA , GROTHEER HORST‐HENNING
Published: March 2007

Abstract:  High levels of ambient air pollution are associated in humans with aggravation of asthma and of respiratory and cardiopulmonary morbidity; long‐term exposures to particulate matter (PM) have been linked to possible increases in lung cancer risk, chronic respiratory disease, and increased death rates. The Biodiagnostics Group of the DLR Institute of Aerospace Medicine develops cellular test systems capable of monitoring the biological consequences of environmental conditions on humans already on cellular and molecular level. Such bioassays rely on the receptor–reporter principle, where cell lines are transfected with plasmids carrying a reporter gene under control of environment‐dependent promoters (receptor), which play a key role in regulating gene expressions in response to extracellular signals. We developed the recombinant human lung epithelial cell line A549‐NF‐κB‐EGFP/Neo carrying a genetically encoded fluorescent indicator for monitoring activation of the NF‐κB signaling pathway in living cells in response to genotoxic and cytotoxic environmental influences. With this cell line we screened several candidate human radiation‐responsive genes (GADD45β, CDKN1A) and NF‐κB‐dependent genes (IL‐6, NFκBIA, and pNF‐κB‐EGFP) for gene expression changes by quantitative reverse transcriptase polymerase chain reaction (qRT‐PCR) assay, using cDNA obtained from total RNA isolated at various time points after exposure to combustion generated nano‐sized particle samples.
DNA Hypomethylation of CAGE Promotors in Squamous Cell Carcinoma of Uterine Cervix
Authors: LEE TAEK SANG , KIM JAE WEON , KANG GYEONG HOON , PARK NOH HYUN , SONG YONG SANG , KANG SOON BEOM , LEE HYO PYO
Published: March 2007

Abstract:  This study was performed to determine whether promotor hypomethylation of CAGE is involved in cervical carcinogenesis. The surgical specimens of 40 cervical squamous cell carcinoma patients treated at Seoul National University Hospital and those of 48 healthy controls were used, with informed consent. We investigated the promotor hypomethylation status of CAGE by methylation‐specific polymerase chain reaction (MSP) using primers specific for unmethylated sequences, and found hypomethylation of CAGE promotor at a frequency approaching 90% in cervical squamous cell carcinomas (35/40, 87.5%), but at less than 4% in controls (P < 0.001). This finding provides experimental evidence of the frequent hypomethylation of normally methylated CAGE promotor CpG islands in cervical cancer, and indicates that this hypomethylation is likely to be a valuable surrogate marker for the expression of CAGE. It also provides a clue concerning the molecular mechanisms of carcinogenesis in cervical squamous cell carcinoma.
The cAMP‐Responsive Unit of the Human Insulin‐Like Growth Factor–Binding Protein‐1 Coinstitutes a Functional Insulin‐Response Element
Authors: SCHWEIZER‐GROYER GHISLAINE , FALLOT GUILLAUME , CADEPOND FRANÇOISE , GIRARD CHRISTELLE , GROYER ANDRÉ
Published: March 2007

Abstract:  Insulin‐like growth factor–binding protein‐1 (IGFBP‐1) is one of the genes involved in glucose homeostasis. In vivo, its level is increased by counter‐regulatory hormones (glucocorticoids and glucagon via its second messenger cAMP) and decreased by insulin, these variations being primarily correlated with IGFBP‐1 gene transcription. Previous reports described a functional insulin response element (IRE), immediately 5′‐ to the glucocorticoid response element (GRE). This IRE has been shown to mediate partial inhibition (1) of basal IGFBP‐1 promoter activity and (2) of glucocorticoid‐induced stimulation of gene transcription by insulin.
In this work, using human HepG2 hepatoma cells as a model system, we showed: (1) that insulin inhibited both basal and cAMP‐induced hIGFBP‐1 promoter (nt‐1 to ‐341) activity; (2) that in the absence of insulin, forkhead box class O (FOXO) transcription factors enhance constitutive hIGFBP‐1 promoter activity without interfering with the stimulatory effect of cAMP; (3) that PI‐3′ kinase signaling is involved in the inhibition of constitutive and cAMP‐induced promoter activities by insulin; (4) that wild‐type FOXO‐1 mediates the inhibitory effect of insulin on the promoter, although FOXO‐1Ala3, a nonphosphorylatable mutant of FOXO‐1, does not; (5) that the cAMP‐responsive unit (CRU), that includes a putative IRE (nt‐265 to ‐282) and a cAMP responsive element (CRE; nt‐258 to ‐263), is sufficient per se to mediate both cAMP stimulation of a heterologous promoter, and inhibition of both basal and cAMP‐induced promoter activities by insulin; and (6) that the inhibitory effects of insulin on the isolated CRU are mediated by the FOXOs. This study is the first evidence for the occurrence of a second IRE within hIGFBP‐1 promoter sequences, IRECRU, located 5′‐ to the CRE.
Altered Gene Expression Pattern in Peripheral Blood Leukocytes from Patients with Arterial Hypertension
Authors: TIMOFEEVA A.V. , GORYUNOVA L.E. , KHASPEKOV G.L. , KOVALEVSKII D.A. , SCAMROV A.V. , BULKINA O.S. , KARPOV YU.A. , TALITSKII K.A. , BUZA V.V. , BRITAREVA V.V. , BEABEALASHVILLI R.SH.
Published: March 2007

Abstract:  The role of various inflammatory mechanisms and oxidative stress in the development of atherosclerosis and arterial hypertension (AH) has been increasingly acknowledged during recent years. Hypertension per se or factors that cause hypertension along with other complications lead to infiltration of activated leukocytes in the vascular wall, where these cells contribute to the development of vascular injury by releasing cytokines, oxygen radicals, and other toxic mediators. However, molecular mechanisms underlying leukocyte activation at transcriptional level in AH are still far from being clear. To solve this problem we employed cDNA microarray technology to reveal the differences in gene expression in peripheral blood leukocytes from patients with AH compared with healthy individuals. The microarray data were verified by a semi‐quantitative RT‐PCR method. We found 25 genes with differential expression in leukocytes from AH patients among which 21 genes were upregulated and 4 genes were downregulated. These genes are implicated in apoptosis (CASP2, CASP4, and CASP8, p53, UBID4, NAT1, and Fte‐1), inflammatory response (CAGC, CXCR4, and CX3CR1), control of MAP kinase function (PYST1, PAC1, RAF1, and RAFB1), vesicular trafficking of molecules among cellular organelles (GDI‐1 and GDI‐2), cell redox homeostasis (GLRX), cellular stress (HSPA8 and HSP40), and other processes. Gene expression pattern of the majority of genes was similar in AH patients independent of the disease stage and used hypotensive therapy, but was clearly different from that of normotensive subjects.
Effects of Histone Deacetylase Inhibitors, Sodium Phenyl Butyrate and Vitamin B3, in Combination with Retinoic Acid on Granulocytic Differentiation of Human Promyelocytic Leukemia HL‐60 Cells
Authors: MERZVINSKYTE RASA , TREIGYTE GRAZINA , SAVICKIENE JURATE , MAGNUSSON KARL‐ERIC , NAVAKAUSKIENE RUTA
Published: March 2007

Abstract:  Water‐soluble vitamin B3, niacin, and its related compounds were suggested to be applicable for medical use. In this article, we examined the anti‐leukemic effects of two distinct histone deacetylase (HDAC1 and Sir2) inhibitors, sodium phenyl butyrate (PB) and vitamin B3, respectively, on human promyelocytic leukemia cells HL‐60, using HDACIs alone and in combination with all trans retinoic acid (RA). We demonstrated that the HDACI combinations exert different effects on cell cycle arrest and differentiation as determined by nitro blue reduction and the expression of the early myeloid differentiation marker CD11b. The most beneficial effects were found by use of 6‐h pretreatment with PB and vitamin B3 before the exposition to RA alone or in combination with vitamin B3, showing significant acceleration and a high level of granulocytic differentiation. The effects were associated with a rapid histone H4 acetylation and later histone H3 modifications. Our results suggest that the use of two HDACI altogether before the induction of differentiation and acting via chromatin remodeling may be promising for the treatment of acute promyelocytic leukemia.
The Histone Deacetylase Inhibitor FK228 Distinctly Sensitizes the Human Leukemia Cells to Retinoic Acid‐Induced Differentiation
Authors: SAVICKIENE JURATE , TREIGYTE GRAZINA , BORUTINSKAITE VERONIKA , NAVAKAUSKIENE RUTA , MAGNUSSON KARL‐ERIC
Published: March 2007

Abstract:  FK228 (depsipeptide) is a novel histone deacetylase inhibitor (HDACI) that has shown therapeutical efficacy in clinical trials for malignant lymphoma. In this article, we examined in vitro effects of FK228 on human leukemia cell lines, NB4 and HL‐60. FK228 alone (0.2–1 ng/mL) inhibited leukemia cell growth in a dose‐dependent manner and induced death by apoptosis. FK228 had selective differentiating effects on two cell lines when used for 6 h before induction of granulocytic differentiation by retinoic acid (RA) or in combination with RA. These effects were accompanied by a time‐ and dose‐dependent histone H4 hyper‐acetylation or histone H3 dephosphorylation and alterations in DNA binding of NF‐κB in association with cell death and differentiation. Pifithrin‐α (PFT), an inhibitor of p53 transcriptional activity, protected only NB4 cells with functional p53 from FK228‐induced apoptosis and did not interfere with antiproliferative activity in p53‐negative HL‐60 cells. In NB4 cells, PFT inhibited p53 binding to the p21 (Waf1/Cip1) promotor and induced DNA binding of NF‐κB leading to enhanced cell survival. Thus, beneficial effects of FK228 on human promyelocytic leukemia may be exerted through the induction of differentiation or apoptosis via histone modification and selective involvement of transcription factors, such as NF‐κB and p53.
Protein Folding Information in Nucleic Acids which Is Not Present in the Genetic Code
Authors: BIRO JAN C.
Published: March 2007

Abstract:  Nucleic acid subsequences comprising the 1st and/or 3rd codon residues in mRNAs express significantly higher free folding energy (FFE) than the subsequence containing only the 2nd residues (P < 0.0001, n= 81). This periodic FFE difference is not present in introns. The FFE in the 1st and 3rd residues is additive, which suggests that these residues contain a significant number of complementary bases and contribute to selection for local mRNA secondary structures. This periodic, codon‐related structure forming of mRNAs indicates a connection between the structure of exons and the corresponding (translated) proteins. The folding energy dot plots of RNAs and the residue contact maps of the coded proteins are indeed similar. Residue contact statistics using 81 different protein structures confirmed that amino acids that are coded by partially reverse and complementary codons (Watson–Crick base pairs at the 1st and 3rd codon positions and translated in reverse orientation) are preferentially colocated in protein structures.
Antigens and Cytokine Genes in Antitumor Vaccines
Authors: HERRERO MARÍA JOSÉ , BOTELLA RAFAEL , DASÍ FRANCISCO , ALGÁS ROSA , SÁNCHEZ MARÍA , ALIÑO SALVADOR F.
Published: March 2007

Abstract:  Studies against cancer, including clinical trials, have shown that a correct activation of the immune system can lead to tumor rejection whereas incorrect signaling results in no positive effects or even anergy. We have worked assuming that two signals, GM‐CSF (granulocyte and macrophage colony‐stimulating factor) and tumor antigens are necessary to mediate an antitumor effective response. To study which is the ideal temporal sequence for their administration, we have used a murine model of antimelanoma vaccine employing whole B16 tumor cells or their membrane protein antigens (TMPs) in combination with gm‐csf transfer before or after the antigen delivery. Our results show that: (i) When gm‐csf tisular transfection is performed before TMP delivery, a tumor growth inhibition is observed, but with a limit effect when administering high antigen doses; in contrast, when signals are inverted, the limited effect is lost and greater antitumor efficacy is obtained. (ii) A similar behavior, but with stronger positive results, is observed employing gm‐csf transfection and whole tumor cells as antigens. While negative results are obtained with gm‐csf before cells, the best results (total survival of treated mice) are obtained when GM‐CSF is administered in transfected cells. We conclude that optimal antitumoral response can be obtained when the antigen signal is given before (or simultaneous with) GM‐CSF production, while the inversion of the signals could result in the undesired inhibition or anergy of the immune response.
Process Simulation in a Mechatronic Bioreactor Device with Speed‐Regulated Motors for Growing of Three‐Dimensional Cell Cultures
Authors: MIHAILOVA MINA , TRENEV VASSIL , GENOVA PENKA , KONSTANTINOV SPIRO
Published: March 2007

Abstract:  Tissue engineering is a new scientific research field that allows the establishment of tissue equivalents rising from isolated cells in combination with biocompatible materials and cultivation in more or less sophisticated bioreactor systems. Such systems gave the unique opportunity to perform in vitro investigations of transcription and translation, cell growth, biochemistry and mechanics of healthy normal organs as well as those affected by malignant tumors, infections, and immune deficiency under controlled conditions. In rotating vessel bioreactors under microgravity and defined medium content, cells proliferate, stay abundant to each other, and form three‐dimensional structures, assigned as spheroids. Such spheroids might be grown on microcarriers. A wide spectrum of different cell culture experiments involving normal and transformed human cells indicates that: in the rotating bioreactor system miniPERM no complete lack of gravity could be reached; a great part of the seeded cell material does not proliferate at the beginning; and the appearance of bigger spheroids is rather random. We describe the acquisition of spheroids from HD‐MY‐Z and Neuro‐2A tumor cells. Spheroids of 100 and more cells were obtained from HD‐MY‐Z and Neuro‐2A cells. Interestingly, chronic myeloid leukemia LAMA‐84 cells did not form any cell clumps and they kept a completely undifferentiated phenotype despite their semiadherent manner of growth under conventional conditions. A detailed theoretical and virtual simulation study of the influence of every component of gravitation, inertia, and hydrodynamic force fields was performed. Therefore, a new concept for mechatronic bioreactor device with active electronic control was developed and virtually tested.
Dual Activity of Phosphorothioate CpG Oligodeoxynucleotides on HIV
Authors: SCHELLER CARSTEN , ULLRICH ANETT , LAMLA STEFAN , DITTMER ULF , RETHWILM AXEL , KOUTSILIERI ELENI
Published: March 2007

Abstract:  CpG oligodeoxynucleotides (CpG ODNs) bind to toll‐like receptor‐9 (TLR‐9) and activate immune cells with antigen‐presenting activity, including B cells and dendritic cells. Here we show that treatment of the latently human immunodeficiency virus (HIV)‐infected T cell line ACH‐2 with the CpG ODNs 2006 or 2040 triggers activation of viral gene expression, demonstrating that CpG‐signaling activity can also be found in T cells. The CpG ODNs g12AAC and g12GTC had no effect on virus reactivation. In contrast to the stimulating effects on viral gene expression in latently infected cells, CpG ODNs potently suppressed HIV replication in productively infected MT4 T cells or PBLs. Inhibition of virus replication was not related to the CpG motif but similarly occurred with non‐CpG phosphorothioate (PTO)‐ODNs. Thus, virus inhibition was likely caused by the PTO backbone of the CpG ODNs, probably by interfering with events prior to integration of the viral cDNA into the host genome. The ability of CpG PTO‐ODNs to trigger reactivation of latent HIV in combination with their antiviral activity on productive infection makes this substance class an interesting candidate for further test to asses their potential as supplements in HIV therapy.
Electron Transfer Reactions in Laser Desorption/Ionization and Matrix-Assisted Laser Desorption/Ionization: Factors Influencing Matrix and Analyte Ion Intensities
Authors: Hoteling Andrew J. , Nichols William F. , Giesen David J. , Lenhard Jerome R. , Knochenmuss Richard
Published: March 2007

Certain analytes appear in laser desorption/ionization or matrix-assisted laser desorption/ionization (MALDI) mass spectra as radical ions, rather than deprotonated, protonated or cationized molecules. Radical ion intensity ratios are found to be consistent with matrix–analyte and analyte–analyte electron transfer (ET) reactions in the desorption/ablation plume. As predicted by the two-step model of ultraviolet MALDI, both suppression of matrix by analytes, and one analyte by another are observed. The dependence of these effects on mixing ratios, laser fluence and matrix choice parallel suppression phenomena previously observed with proton or cation transfer reactions are discussed. The driving force for ET, as reflected in differences of gas-phase molecular ionization potentials (IPs) or solution oxidation potentials (Eox), is usually the dominant factor determining relative ion intensitites. In positive polarity, low IP (Eox) analytes suppress signals of high IP (Eox) analytes. However, for large IP (Eox) differences and at high matrix-to-analyte mole ratios, relative intensities were found to invert. This is tentatively ascribed to a rollover in the rate of matrix–analyte electron transfer at high exoergicity.
Predicting the intention to quit smoking and quitting behaviour: Extending the theory of planned behaviour
Authors: Rise J. , Kovac V. , Kraft P. , Moan I. S.
Published: March 2007

Objectives. The present study examined the ability of the TPB to predict the intention to quit smoking and quitting behaviour. In addition, the predictive power of future orientation, number of cigarettes smoked, planning, past behaviour and the interactions between intention and other predictors was examined.

Material and methods. The data were derived from a longitudinal survey among 103 daily smoking students at the University of Oslo (mean age 24.6 years, mean number of years of smoking = 8 years). These data were collected by means of self‐administered questionnaires at T1 (October 2003) and at T2 (February 2004) in terms of recording actual quitting.

Results. The TPB components accounted for 30% of the variance in quitting intentions, and affective attitude and descriptive norm emerged as the strongest predictors of quitting intention. Ordinal regression analysis showed that intention was a borderline significant predictor of subsequent quitting behaviour, while the impact of PBC was non‐significant (model 1). The inclusion of the additional variables improved the fit of the model, with number of cigarettes and planning appearing as significant predictors of behaviour (model 2). As predicted, there was a significant interaction between perceived control and intentions on quitting (model 3). Nagelkerke R2 increased from .07 in model 1 to .54 in model 2, and finally to .58 in model 3.

Conclusions. The results indicate that affective attitude and descriptive norm play a more crucial role than the other TPB predictors in motivating smokers to quit. The results also indicate that self‐regulatory strategies are important in relation to addictive behaviours.
Specific Defects in Different Transcription Complexes Compensate for the Requirement of the Negative Cofactor 2 Repressor in Saccharomyces cerevisiae
Authors: Peiró-Chova Lorena , Estruch Francisco
Published: March 2007

Negative cofactor 2 (NC2) has been described as an essential and evolutionarily conserved transcriptional repressor, although in vitro and in vivo experiments suggest that it can function as both a positive and a negative effector of transcription. NC2 operates by interacting with the core promoter and components of the basal transcription machinery, like the TATA-binding protein (TBP). In this work, we have isolated mutants that suppress the growth defect caused by the depletion of NC2. We have identified mutations affecting components of three different complexes involved in the control of basal transcription: the mediator, TFIIH, and RNA pol II itself. Mutations in RNA pol II include both overexpression of truncated forms of the two largest subunits (Rpb1 and Rpb2) and reduced levels of these proteins. Suppression of NC2 depletion was also observed by reducing the amounts of the mediator essential components Nut2 and Med7, as well as by deleting any of the nonessential mediator components, except Med2, Med3, and Gal11 subunits. Interestingly, the Med2/Med3/Gal11 triad forms a submodule within the mediator tail. Our results support the existence of different components within the basic transcription complexes that antagonistically interact with the NC2 repressor and suggest that the correct balance between the activities of specific positive and negative components is essential for cell growth.
Genetic Similarities Within and Between Human Populations
Authors: Witherspoon D J , Wooding S , Rogers A R , Marchani E E , Watkins W S , Batzer M A , Jorde L B
Published: March 2007

The proportion of human genetic variation due to differences between populations is modest, and individuals from different populations can be genetically more similar than individuals from the same population. Yet sufficient genetic data can permit accurate classification of individuals into populations. Both findings can be obtained from the same data set, using the same number of polymorphic loci. This article explains why. Our analysis focuses on the frequency, ω, with which a pair of random individuals from two different populations is genetically more similar than a pair of individuals randomly selected from any single population. We compare ω to the error rates of several classification methods, using data sets that vary in number of loci, average allele frequency, populations sampled, and polymorphism ascertainment strategy. We demonstrate that classification methods achieve higher discriminatory power than ω because of their use of aggregate properties of populations. The number of loci analyzed is the most critical variable: with 100 polymorphisms, accurate classification is possible, but ω remains sizable, even when using populations as distinct as sub-Saharan Africans and Europeans. Phenotypes controlled by a dozen or fewer loci can therefore be expected to show substantial overlap between human populations. This provides empirical justification for caution when using population labels in biomedical settings, with broad implications for personalized medicine, pharmacogenetics, and the meaning of race.
Precise Mapping of Quantitative Trait Loci for Resistance to Southern Leaf Blight, Caused by Cochliobolus heterostrophus Race O, and Flowering Time Using Advanced Intercross Maize Lines
Authors: Balint-Kurti P J , Zwonitzer J C , Wisser R J , Carson M L , Oropeza-Rosas M A , Holland J B , Szalma S J
Published: March 2007

The intermated B73 × Mo17 (IBM) population, an advanced intercross recombinant inbred line population derived from a cross between the maize lines B73 (susceptible) and Mo17 (resistant), was evaluated in four environments for resistance to southern leaf blight (SLB) disease caused by Cochliobolus heterostrophus race O. Two environments were artificially inoculated, while two were not inoculated and consequently had substantially lower disease pressure. Four common SLB resistance quantitative trait loci (QTL) were identified in all environments, two in bin 3.04 and one each in bins 1.10 and 8.02/3. There was no significant correlation between disease resistance and days to anthesis. A direct comparison was made between SLB QTL detected in two populations, independently derived from the same parental cross: the IBM advanced intercross population and a conventional recombinant inbred line population. Several QTL for SLB resistance were detected in both populations, with the IBM providing between 5 and, in one case, 50 times greater mapping resolution.
Comparative Physical Mapping Between Oryza sativa (AA Genome Type) and O. punctata (BB Genome Type)
Authors: Kim HyeRan , Miguel Phillip San , Nelson William , Collura Kristi , Wissotski Marina , Walling Jason G , Kim Jun Pyo , Jackson Scott A , Soderlund Carol , Wing Rod A
Published: March 2007

A comparative physical map of the AA genome (Oryza sativa) and the BB genome (O. punctata) was constructed by aligning a physical map of O. punctata, deduced from 63,942 BAC end sequences (BESs) and 34,224 fingerprints, onto the O. sativa genome sequence. The level of conservation of each chromosome between the two species was determined by calculating a ratio of BES alignments. The alignment result suggests more divergence of intergenic and repeat regions in comparison to gene-rich regions. Further, this characteristic enabled localization of heterochromatic and euchromatic regions for each chromosome of both species. The alignment identified 16 locations containing expansions, contractions, inversions, and transpositions. By aligning 40% of the punctata BES on the map, 87% of the punctata FPC map covered 98% of the O. sativa genome sequence. The genome size of O. punctata was estimated to be 8% larger than that of O. sativa with individual chromosome differences of 1.5–16.5%. The sum of expansions and contractions observed in regions >500 kb were similar, suggesting that most of the contractions/expansions contributing to the genome size difference between the two species are small, thus preserving the macro-collinearity between these species, which diverged ∼2 million years ago.
Neurospora Spore KillersSk-2andSk-3Suppress Meiotic Silencing by Unpaired DNA
Authors: Raju Namboori B , Metzenberg Robert L , Shiu Patrick K T
Published: March 2007

In Neurospora crassa, pairing of homologous DNA segments is monitored during meiotic prophase I. Any genes not paired with a homolog, as well as any paired homologs of that gene, are silenced during the sexual phase by a mechanism known as meiotic silencing by unpaired DNA (MSUD). Two genes required for MSUD have been described previously: sad-1 (suppressor of ascus dominance), encoding an RNA-directed RNA polymerase, and sad-2, encoding a protein that controls the perinuclear localization of SAD-1. Inactivation of either sad-1 or sad-2 suppresses MSUD. We have now shown that MSUD is also suppressed by either of two Spore killer strains, Sk-2 and Sk-3. These were both known to contain a haplotype segment that behaves as a meiotic drive element in heterozygous crosses of killer × sensitive. Progeny ascospores not carrying the killer element fail to mature and are inviable. Crosses homozygous for either of the killer haplotypes suppress MSUD even though ascospores are not killed. The killer activity maps to the same 30-unit-long region within which recombination is suppressed in killer × sensitive crosses. We suggest that the region contains a suppressor of MSUD.