ECONOMIC HISTORY REVIEW, Issue 2 2009
STEPHEN BROADBERRY
In the early eighteenth century, wages in Britain were more than four times as high as in India, the world's major exporter of cotton textiles. This induced the adoption of more capital-intensive production methods in Britain and a faster rate of technological progress, so that competitive advantage had begun to shift in Britain's favour by the late eighteenth century. However, the completion of the process was delayed until after the Napoleonic Wars by increasing raw cotton costs, before supply adjusted to the major increase in demand for inputs. [source]

High-efficiency peptide analysis on monolithic multimode capillary columns: Pressure-assisted capillary electrochromatography/capillary electrophoresis coupled to UV and electrospray ionization-mass spectrometry

ELECTROPHORESIS, Issue 21 2003
Alexander R. Ivanov
Abstract High-efficiency peptide analysis using multimode pressure-assisted capillary electrochromatography/capillary electrophoresis (pCEC/pCE) monolithic polymeric columns and the separation of model peptide mixtures and protein digests by isocratic and gradient elution under an applied electric field with UV and electrospray ionization-mass spectrometry (ESI-MS) detection is demonstrated. Capillary multipurpose columns were prepared in silanized fused-silica capillaries of 50, 75, and 100 ,m inner diameters by thermally induced in situ copolymerization of methacrylic monomers in the presence of n -propanol and formamide as porogens and azobisisobutyronitrile as initiator. N -Ethylbutylamine was used to modify the chromatographic surface of the monolith from neutral to cationic. Monolithic columns were termed as multipurpose or multimode columns because they showed mixed modes of separation mechanisms under different conditions. Anion-exchange separation ability in the liquid chromatography (LC) mode can be determined by the cationic chromatographic surface of the monolith. At acidic pH and high voltage across the column, the monolithic stationary phase provided conditions for predominantly capillary electrophoretic migration of peptides. At basic pH and electric field across the column, enhanced chromatographic retention of peptides on monolithic capillary column made CEC mechanisms of migration responsible for separation. The role of pressure, ionic strength, pH, and organic content of the mobile phase on chromatographic performance was investigated. High efficiencies (exceeding 300,000 plates/m) of the monolithic columns for peptide separations are shown using volatile and nonvolatile, acidic and basic buffers. Good reproducibility and robustness of isocratic and gradient elution pressure-assisted CEC/CE separations were achieved for both UV and ESI-MS detection. Manipulation of the electric field and gradient conditions allowed high-throughput analysis of complex peptide mixtures. A simple design of sheathless electrospray emitter provided effective and robust low dead volume interfacing of monolithic multimode columns with ESI-MS. Gradient elution pressure-assisted mixed-mode separation CE/CEC-ESI-MS mass fingerprinting and data-dependent pCE/pCEC-ESI-MS/MS analysis of a bovine serum albumin (BSA) tryptic digest in less than 5 min yielding high sequence coverage (73%) demonstrated the potential of the method. [source]

Proteomic analysis of cellular responses to low concentration N -methyl- N,-nitro- N -nitrosoguanidine in human amnion FL cells

ENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 2 2004
Jinghua Jin
Abstract We have shown previously that exposure to a low concentration of N -methyl- N,-nitro- N -nitrosoguanidine (MNNG) induces comprehensive changes in the protein expression profile of human amnion FL cells, including the induction, suppression, upregulation, and downregulation of various proteins. In addition, by proteomic analysis combining two-dimensional gel electrophoresis (2-DE) and mass spectrometry, some of the induced and suppressed proteins were identified. In this study, we identified an additional 18 proteins among those that were either up- or downregulated by MNNG treatment. The proteins identified were a heterogeneous group that included several zinc finger proteins, proteins involved in signal transduction, cytoskeletal proteins, cell-cycle regulation proteins, and proteins with unknown functions. The involvement of these proteins in the cellular responses to alkylating agents has not been reported before and their physiological relevance is not clear. Therefore, our findings may help better understand the global cellular stress responses to chemical carcinogens, and may lead to new studies on the functions of these MNNG-responsive proteins. Furthermore, some of these proteins may serve as biomarkers for detecting exposure of human populations to environmental carcinogens. Environ. Mol. Mutagen. 43:93,99, 2004. © 2004 Wiley-Liss, Inc. [source]

Use of Chronic Epilepsy Models in Antiepileptic Drug Discovery: The Effect of Topiramate on Spontaneous Motor Seizures in Rats with Kainate-induced Epilepsy

EPILEPSIA, Issue 1 2005
Heidi L. Grabenstatter
Summary:,Purpose: Potential antiepileptic drugs (AEDs) are typically screened on acute seizures in normal animals, such as those induced in the maximal electroshock and pentylenetet-razole models. As a proof-of-principle test, the present experiments used spontaneous epileptic seizures in kainate-treated rats to examine the efficacy of topiramate (TPM) with a repeated-measures, crossover protocol. Methods: Kainic acid was administered in repeated low doses (5 mg/kg) every hour until each Sprague,Dawley rat experienced convulsive status epilepticus for >3 h. Six 1-month trials (n = 6,10 rats) assessed the effects of 0.3,100 mg/kg TPM on spontaneous seizures. Each trial involved six pairs of TPM and saline-control treatments administered as intraperitoneal injections on alternate days with a recovery day between each treatment day. Data analysis included a log transformation to compensate for the asymmetric distribution of values and the heterogeneous variances, which appeared to arise from clustering of seizures. Results: A significant effect of TPM was observed for 12 h (i.e., two 6-h periods) after a 30-mg/kg injection, and full recovery from the drug effect was complete within 43 h. TPM exerted a significant effect at doses of 10, 30, and 100 mg/kg, and the effects of TPM (0.3,100 mg/kg) were dose dependent. Conclusions: These data suggest that animal models with spontaneous seizures, such as kainate- and pilocarpine-treated rats, can be used efficiently for rapid testing of AEDs with a repeated-measures, crossover protocol. Furthermore, the results indicate that this design allows both dose,effect and time-course-of-recovery studies. [source]

RelB/p50 regulates CCL19 production, but fails to promote human DC maturation

EUROPEAN JOURNAL OF IMMUNOLOGY, Issue 8 2009
Chiara Gasparini
Abstract DC, when fully matured are the APC best able to activate naïve T cells. Recently, we demonstrated using adenoviruses overexpressing I,B, and proteosome inhibitors that NF-,B is involved in DC activation, but the role of the individual subunits is still not clear. We investigated the role of the NF-,B subunits RelB and p50 in human DC activation using adenoviral vectors expressing RelB or p50. Nuclear RelB, in the form of RelB/p50, was active only in DC infected with both viruses, this induced the production of the soluble homeostatic chemokine CCL19, but not other homeostatic chemokines, particularly in LPS-matured DC. However, RelB/p50 did not affect the expression of costimulatory and antigen-presenting molecules, and increased the allogeneic mixed lymphocyte reaction only in LPS-matured DC. This enhanced mixed lymphocyte reaction is most likely due to enhanced CCL19 production, which sustains the interaction between mature DC and naïve T cells. In conclusion, we demonstrated that RelB/p50 was active only in DC expressing both RelB and p50, and induced CCL19 production, but not DC maturation. [source]

Empathy-induced altruism in a prisoner's dilemma II: what if the target of empathy has defected?

EUROPEAN JOURNAL OF SOCIAL PSYCHOLOGY, Issue 1 2001
C. Daniel Batson
What if participants in a one-trial prisoner's dilemma know before making their decision that the other person has already defected? From the perspective of classic game theory, a dilemma no longer exists. It is clearly in their best interest to defect too. The empathy-altruism hypothesis predicts, however, that if they feel empathy for the other, then a dilemma remains: self-interest counsels defection; empathy-induced altruism counsels not. This motivational conflict should lead at least some empathically aroused individuals not to defect. To test this prediction, we placed 60 undergraduate women in a one-trial prisoner's dilemma in which they knew the other had already defected. Among those not induced to feel empathy, very few (0.05) did not defect in return. Among those induced to feel empathy for the other, almost half (0.45) did not defect. These results underscore the power of empathy-induced altruism to affect responses in a prisoner's dilemma. Copyright © 2001 John Wiley & Sons, Ltd. [source]

Molecular monitoring of microbial diversity in expanded granular sludge bed (EGSB) reactors treating oleic acid

FEMS MICROBIOLOGY ECOLOGY, Issue 2 2002
Maria Alcina Pereira
Abstract A molecular approach was used to evaluate the microbial diversity of bacteria and archaea in two expanded granular sludge bed (EGSB) reactors fed with increasing oleic acid loading rates up to 8 kg of chemical oxygen demand (COD) m,3 day,1 as the sole carbon source. One of the reactors was inoculated with granular sludge (RI) and the other with suspended sludge (RII). During operation, the sludge in both reactors was segregated in two layers: a bottom settled one and a top floating one. The composition of the bacterial community, based on 16S rDNA sequence diversity, was affected most during the oleate loading process in the two reactors. The archaeal consortium remained rather stable over operation in RI, whereas in RII the relative abundance of Methanosaeta -like organisms became gradually weaker, starting in the bottom layer. In the range of oleate loads evaluated, 6 kg of COD m,3 day,1 was found as the maximum value that could be applied to the system. A further increase to 8 kg of oleate-COD m,3 day,1 induced a maximal shift on the microbial structure of the sludges. At this time point, methanogenic acetoclastic activity was not detected and only very low methanogenic activity on H2/CO2 was exhibited by the sludges. [source]

Beta-amyloid peptide stimulates endozepine release in cultured rat astrocytes through activation of N -formyl peptide receptors

GLIA, Issue 13 2008
Tursonjan Tokay
Abstract Astroglial cells synthesize and release endozepines, a family of neuropeptides derived from diazepam-binding inhibitor (DBI). The authors have recently shown that ,-amyloid peptide (A,) stimulates DBI gene expression and endozepine release. The purpose of this study was to determine the mechanism of action of A, in cultured rat astrocytes. A,25,35 and the N -formyl peptide receptor (FPR) agonist N -formyl-Met-Leu-Phe (fMLF) increased the secretion of endozepines in a dose-dependent manner with EC50 value of ,2 ,M. The stimulatory effects of A,25,35 and the FPR agonists fMLF and N -formyl-Met-Met-Met (fMMM) on endozepine release were abrogated by the FPR antagonist N - t -Boc-Phe-Leu-Phe-Leu-Phe. In contrast, A,25,35 increased DBI mRNA expression through a FPR-independent mechanism. A,25,35 induced a transient stimulation of cAMP formation and a sustained activation of polyphosphoinositide turnover. The stimulatory effect of A,25,35 on endozepine release was blocked by the adenylyl cyclase inhibitor somatostatin, the protein kinase A (PKA) inhibitor H89, the phospholipase C inhibitor U73122, the protein kinase C (PKC) inhibitor chelerythrine and the ATP binding cassette transporter blocker glyburide. Taken together, these data demonstrate for the first time that A,25,35 stimulates endozepine release from rat astrocytes through a FPR receptor positively coupled to PKA and PKC. © 2008 Wiley-Liss, Inc. [source]

Transforming growth factor-activated kinase 1 induced in spinal astrocytes contributes to mechanical hypersensitivity after nerve injury

GLIA, Issue 7 2008
Hirokazu Katsura
Abstract Mitogen-activated protein kinase (MAPK) plays an important role in the induction and maintenance of neuropathic pain. Transforming growth factor-activated kinase 1 (TAK1), a member of the MAPK kinase kinase family, is indispensable for the activation of c-Jun N-terminal kinase (JNK) and p38 MAPK. We now show that TAK1 induced in spinal cord astrocytes is crucial for mechanical hypersensitivity after peripheral nerve injury. Nerve injury induced a striking increase in the expression of TAK1 in the ipsilateral dorsal horn, and TAK1 was increased in hyperactive astrocytes, but not in neurons or microglia. Intrathecal administration of TAK1 antisense oligodeoxynucleotide (AS-ODN) prevented and reversed nerve injury-induced mechanical, but not heat hypersensitivity. Furthermore, TAK1 AS-ODN suppressed the activation of JNK1, but not p38 MAPK, in spinal astrocytes. In contrast, there was no change in TAK1 expression in primary sensory neurons, and TAK1 AS-ODN did not attenuate the induction of transient receptor potential ion channel TRPV1 in sensory neurons. Taken together, these results demonstrate that TAK1 upregulation in spinal astrocytes has a substantial role in the development and maintenance of mechanical hypersensitivity through the JNK1 pathway. Thus, preventing the TAK1/JNK1 signaling cascade in astrocytes might provide a fruitful strategy for treating intractable neuropathic pain. © 2008 Wiley-Liss, Inc. [source]

Rationalization of the stereochemistry of an addition of dialkyl phosphites to certain chiral aldimines: The experimental and theoretical approach

HETEROATOM CHEMISTRY, Issue 2 2002
Ryszard B. Nazarski
The absolute configuration of an ,-P stereogenic center in two diastereomeric O,O-dialkyl ,-aminophosphonates (3), arising from an induced 1,3-asymmetric phosphite addition to the CN bond of furfural-derived Schiff bases (1), was established from single product 1H NMR data. Such spectra were interpreted with anisotropic shielding in relation to the AM1 and MNDO/d structures of 3; the former ones turned out to be closer to the obtained experimental results (1H NMR spectra of 3, crystallographic database study). Since favored 3-21G geometries of starting imines 1 were modeled as well, it was inferred that a stereochemical outcome of this reaction is governed by Cram selectivity. © 2002 Wiley Periodicals, Inc. Heteroatom Chem 13:120,125, 2002; Published online in Wiley Interscience (www.interscience.wiley.com). DOI 10.1002/hc.10005 [source]

Induced and repressed genes after irradiation sensitizing by pentoxyphylline,

INTERNATIONAL JOURNAL OF CANCER, Issue 6 2007
Waldemar Waldeck
Abstract Aim in cancer therapy is to increase the therapeutic ratio eliminating the disease while minimizing toxicity to normal tissues. Radiation therapy is a main component in targeting cancer. Radiosensitizing agents like pentoxyphylline (PTX) have been evaluated to improve radiotherapy. Commonly, cells respond to radiation by the activation of specific early and late response genes as well as by inhibition of genes, which are expressed under normal conditions. A display of the genetic distinctions at the level of transcription is given here to characterize the molecular events underlying the radiosensitizing mechanisms. The method of suppression subtractive hybridization allows the visualization of both induced and repressed genes in irradiated cells compared with cells sensitized immediately after irradiation. The genes were isolated by cDNA-cloning, differential analysis and sequence similarity search. Genes involved in protein synthesis, metabolism, proteolysis and transcriptional regulation were detected. It is important that genes like KIAA280, which were only known as unidentified EST sequences before without function, but inaccessible by array technology were recovered as functional genes. Database searches for PTX-induced genes detected a human mRNA completely unknown. In case of suppressed genes, we detected several mRNAs; one thereof shows homology to a hypothetical protein possibly involved in signal transduction. A further mRNA encodes the protein BM036 supposed to associate with the E2F transcription factor. A hypothetical protein H41 was detected, which may repress the Her-2/neu receptor influencing breast cancer, gliomas and prostate tumors. Radiation combined with PTX may lead to a better prognosis by down regulation of the Her-2/neu, which will be proven by clinical studies in the near future. © 2006 Wiley-Liss, Inc. [source]

Induced ordered weighted geometric operators and their use in the aggregation of multiplicative preference relations

INTERNATIONAL JOURNAL OF INTELLIGENT SYSTEMS, Issue 3 2004
F. Chiclana
In this article, we introduce the induced ordered weighted geometric (IOWG) operator and its properties. This is a more general type of OWG operator, which is based on the induced ordered weighted averaging (IOWA) operator. We provide some IOWG operators to aggregate multiplicative preference relations in group decision-making (GDM) problems. In particular, we present the importance IOWG (I-IOWG) operator, which induces the ordering of the argument values based on the importance of the information sources; the consistency IOWG (C-IOWG) operator, which induces the ordering of the argument values based on the consistency of the information sources; and the preference IOWG (P-IOWG) operator, which induces the ordering of the argument values based on the relative preference values associated with each one of them. We also provide a procedure to deal with "ties" regarding the ordering induced by the application of one of these IOWG operators. This procedure consists of a sequential application of the aforementioned IOWG operators. Finally, we analyze the reciprocity and consistency properties of the collective multiplicative preference relations obtained using IOWG operators. © 2004 Wiley Periodicals, Inc. [source]

RANKL Treatment Releases the Negative Regulation of the Poly(ADP-Ribose) Polymerase-1 on Tcirg1 Gene Expression During Osteoclastogenesis,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 11 2006
Guillaume E Beranger
Abstract The Tcirg1 gene encodes the osteoclast-specific a3 isoform of the V-ATPase a subunit. Using the mouse osteoclastic model RAW264.7 cells, we studied Tcirg1 gene expression, and we identified PARP-1 as a transcriptional repressor negatively regulated by RANKL during osteoclastogenesis. Introduction: The TCIRG1 gene encodes the a3 isoform of the V-ATPase a subunit, and mutations at this locus account for ,60% of infantile malignant osteopetrosis cases. Using RAW264.7 cells as an osteoclastic differentiation model, we undertook a transcriptional study of the mouse Tcirg1 gene focused on the 4-kb region upstream of the transcription starting point. Materials and Methods: The promoter activity of serial-deletion fragments of the Tcirg1 gene promoter was monitored throughout the RAW264.7 cell differentiation process. We next performed EMSA, UV cross-linking, affinity purification, mass spectrometry analysis, gel supershift, and siRNA transfection experiments to identify the factor(s) interacting with the promoter. Results: The ,3946/+113 region of the mouse Tcirg1 gene displayed a high basal promoter activity, which was enhanced by RANKL treatment of RAW264.7 cells. Constructs deleted up to ,1589 retained this response to RANKL. A deletion up to ,1402 induced a 3-fold enhancement of the basal activity, whereas RANKL response was not affected. EMSA experiments led us to identify within the ,1589/,1402 region, a 10-nucleotide sequence, which bound a nuclear protein present in nondifferentiated RAW264.7 cells. This interaction was lost using nuclear extracts derived from RANKL-treated cells. Affinity purification followed by mass spectrometry analysis and gel supershift assay allowed the identification of poly(ADP-ribose) polymerase-1 (PARP-1) as this transcriptional repressor, whereas Western blot experiments revealed the cleavage of the DNA-binding domain of PARP-1 on RANKL treatment. Finally, both PARP-1 depletion after siRNA transfection and RAW264.7 cell treatment by an inhibitor of PARP-1 activity induced an increase of a3 mRNA expression. Conclusions: We provide evidence that the basal transcription activity of the Tcirg1 gene is negatively regulated by the binding of PARP-1 protein to its promoter region in mouse pre-osteoclast. On RANKL treatment, PARP-1 protein is cleaved and loses its repression effect, allowing an increase of Tcirg1 gene expression that is critical for osteoclast function. [source]

Comparison of Induced and Spontaneous Atrial Tachyarrhythmias in Patients with a History of Spontaneous Atrial Tachyarrhythmias

JOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY, Issue 8 2005
CHRISTIAN G. WOLLMANN M.D.
Introduction: This retrospective study investigated whether induced episodes could be used to predict the morphology of future spontaneous atrial episodes. Methods: Eighty-two patients (64 ± 12 years; 77% male; CAD in 60%; left ventricular ejection fraction 45 ± 16%) with a history of atrial tachycardia or atrial fibrillation (AT/AF) were implanted with a dual-chamber implantable cardioverter defibrillator (ICD) and followed for 6 months. A total of 224 episodes of induced and spontaneous AT/AF were classified into type I, II, and III according to the method of Israel et al. and then compared based on average cycle length (CL) and atrial amplitude. Episodes were also grouped as "pace-terminable" or "nonpace-terminable" based on the CL definition of Gillis et al. Results: The analysis of 121 induced episodes (from 80 patients) and 103 spontaneous episodes (from 43 patients) showed that within each arrhythmia type, there were no significant differences in CL or mean amplitude between induced and spontaneous episodes. Additional analysis of patients that had both induced and spontaneous episodes (n = 41) showed 78% had at least one spontaneous episode that matched the induced episode. Fifty-seven percent of spontaneous episodes were considered to be pace-terminable based on CL. Conclusions: Our data suggest that there is no significant difference between induced and spontaneous episodes of AT/AF of the same type. The majority of patients had at least one spontaneous episode of the same type as the induced episode, showing that induced atrial arrhythmias may be useful in predicting the morphology of future spontaneous episodes and in identifying patients potentially benefiting from atrial antitachycardia pacing. [source]

Nicotine inhibits myofibroblast differentiation in human gingival fibroblasts

JOURNAL OF CELLULAR BIOCHEMISTRY, Issue 6 2005
Yiyu Fang
Abstract Cigarette smoking has been suggested as a risk factor for several periodontal diseases. It has also been found that smokers respond less favorably than non-smokers to periodontal therapy. Previous work in our lab has shown that nicotine inhibits human gingival cell migration. Since myofibroblasts play an important role in wound closure, we asked if nicotine affects gingival wound healing process by regulating myofibroblast differentiation. Human gingival fibroblasts (HGFs) from two patients were cultured in 10% fetal bovine serum cell culture medium. Cells were pretreated with different doses of nicotine (0, 0.01, 0.1, and 1 mM) for 2 h, and then incubated with transforming growth factor beta (TGF-,1) (0, 0.25, 0.5, and 1 ng/ml) with or without nicotine for 30 h. The expression level of ,-smooth muscle actin (,-SMA), a specific marker for myofibroblasts, was analyzed by Western blots, immunocytochemistry, and real-time polymerase chain reaction (real-time PCR). Phosphorylated p38 mitogen-activated protein kinase (Phospho-p38 MAPK) activity was analyzed by Western blots. TGF-,1 induced an increase of ,-SMA protein and mRNA expression, while nicotine (1 mM) inhibited the TGF-,1-induced expression of ,-SMA but not ,-actin. Nicotine treatment down-regulated TGF-,1-induced p38 MAPK phosphorylation. Our results demonstrated for the first time that nicotine inhibits myofibroblast differentiation in human gingival fibroblasts in vitro; supporting the hypothesis that delayed wound healing in smokers may be due to decreased wound contraction by myofibroblasts. © 2005 Wiley-Liss, Inc. [source]

Effect of vaccination with recombinant modified vaccinia virus Ankara expressing structural and regulatory genes of SIVmacJ5 on the kinetics of SIV replication in cynomolgus monkeys

JOURNAL OF MEDICAL PRIMATOLOGY, Issue 4 2001
Donatella R.M. Negri
The efficacy of a multicomponent vaccination with modified vaccinia Ankara constructs (rMVA) expressing structural and regulatory genes of simian immunodeficiency virus (SIVmac251/32H/J5) was investigated in cynomolgus monkeys, following challenge with a pathogenic SIV. Vaccination with rMVA-J5 performed at week 0, 12, and 24 induced a moderate proliferative response to whole SIV, a detectable humoral response to all but Nef SIV antigens, and failed to induce neutralizing antibodies. Two months after the last boost, the monkeys were challenged intravenously with 50 MID50 of SIVmac251. All control monkeys, previously inoculated with non-recombinant MVA, were infected by week two and seroconverted by weeks four to eight. In contrast a sharp increase of both humoral and proliferative responses at two weeks post-challenge was observed in vaccinated monkeys compared to control monkeys. Although all vaccinated monkeys were infected, vaccination with rMVA-J5 appeared to partially control viral replication during the acute and late phase of infection as judged by cell- and plasma-associated viral load. [source]

Germinal epithelium, folliculogenesis, and postovulatory follicles in ovaries of rainbow trout, Oncorhynchus mykiss (Walbaum, 1792) (Teleostei, protacanthopterygii, salmoniformes)

JOURNAL OF MORPHOLOGY, Issue 4 2007
Harry J. Grier
Abstract The rainbow trout, Oncorhynchus mykiss (Walbaum, 1792), is a salmoniform fish that spawns once per year. Ripe females that had ovulated naturally, and those induced to ovulate using salmon gonadotropin-releasing hormone, were studied to determine whether follicles were forming at the time of spawning and to describe the process of folliculogenesis. After ovulation, the ovaries of postspawned rainbow trout were examined histologically, using the periodic acid-Schiff procedure, to stain basement membranes that subtend the germinal epithelium and to interpret and define the activity of the germinal epithelium. After spawning, the ovary contained a few ripe oocytes that did not ovulate, numerous primary growth oocytes including oocytes with cortical alveoli, and postovulatory follicles. The germinal epithelium was active in postspawned rainbow trout, as determined by the presence of numerous cell nests, composed of oogonia, mitotic oogonia, early diplotene oocytes, and prefollicle cells. Cell nests were separated from the stroma by a basement membrane continuous with that subtending the germinal epithelium. Furthermore, follicles containing primary growth oocytes were connected to the germinal epithelium; the basement membrane surrounding the follicle joined that of the germinal epithelium. After ovulation, the basement membrane of the postovulatory follicle was continuous with that of the germinal epithelium. We observed consistent separation of the follicle, composed of an oocyte and surrounding follicle cells, from the ovarian stroma by a basement membrane. The follicle is derived from the germinal epithelium. As with the germinal epithelium, follicle cells derived from it never contact those of the connective tissue stroma. As with epithelia, they are always separated from connective tissue by a basement membrane. J. Morphol., 2007. © 2007 Wiley-Liss, Inc. [source]

Cerebral ischemia/stroke and small ubiquitin-like modifier (SUMO) conjugation , a new target for therapeutic intervention?

JOURNAL OF NEUROCHEMISTRY, Issue 3 2008
Wei Yang
Abstract Transient cerebral ischemia/stroke activates various post-translational protein modifications such as phosphorylation and ubiquitin conjugation that are believed to play a major role in the pathological process triggered by an interruption of blood supply and culminating in cell death. A new system of post-translational protein modification has been identified, termed as small ubiquitin-like modifier (SUMO) conjugation. Like ubiquitin, SUMO is conjugated to the lysine residue of target proteins in a complex process. This review summarizes observations from recent experiments focusing on the effect of cerebral ischemia on SUMO conjugation. Transient global and focal cerebral ischemia both induced a rapid, dramatic and long-lasting rise in levels of SUMO2/3 conjugation. After transient focal cerebral ischemia, SUMO conjugation was particularly prominent in neurons located at the border of the ischemic territory where SUMO-conjugated proteins translocated to the nucleus. Many SUMO conjugation target proteins are transcription factors and sumoylation has been shown to have a major impact on the activity, stability, and cellular localization of target proteins. The rise in levels of SUMO-conjugated proteins is therefore likely to have a major effect on the fate of post-ischemic neurons. The sumoylation process could provide an exciting new target for therapeutic intervention. [source]

Pyroglutamate-modified amyloid ,-peptides , A,N3(pE) , strongly affect cultured neuron and astrocyte survival

JOURNAL OF NEUROCHEMISTRY, Issue 6 2002
Claudio Russo
Abstract N-terminally truncated amyloid-, (A,) peptides are present in early and diffuse plaques of individuals with Alzheimer's disease (AD), are overproduced in early onset familial AD and their amount seems to be directly correlated to the severity and the progression of the disease in AD and Down's syndrome (DS). The pyroglutamate-containing isoforms at position 3 [A,N3(pE),40/42] represent the prominent form among the N-truncated species, and may account for more than 50% of A, accumulated in plaques. In this study, we compared the toxic properties, fibrillogenic capabilities, and in vitro degradation profile of A,1,40, A,1,42, A,N3(pE),40 and A,N3(pE),42. Our data show that fibre morphology of A, peptides is greatly influenced by the C-terminus while toxicity, interaction with cell membranes and degradation are influenced by the N-terminus. A,N3(pE),40 induced significantly more cell loss than the other species both in neuronal and glial cell cultures. Aggregated A,N3(pE) peptides were heavily distributed on plasma membrane and within the cytoplasm of treated cells. A,N3(pE),40/42 peptides showed a significant resistance to degradation by cultured astrocytes, while full-length peptides resulted partially degraded. These findings suggest that formation of N-terminally modified peptides may enhance ,-amyloid aggregation and toxicity, likely worsening the onset and progression of the disease. [source]

Chronic Morphine Treatment and Withdrawal Increase Extracellular Levels of Norepinephrine in the Rat Bed Nucleus of the Stria Terminalis

JOURNAL OF NEUROCHEMISTRY, Issue 2 2000
José Antonio Fuentealba
Extracellular levels of norepinephrine (NE) and glutamate (Glu) in the ventral bed nucleus of the stria terminalis (vBNST) of saline- and chronic morphine-treated rats, with or without withdrawal, were studied by means of the in vivo microdialysis technique in anesthetized rats. In addition, the tissue concentration of NE was studied at different rostrocaudal levels of the vBNST. Chronic morphine treatment significantly increased extracellular levels of NE, but not Glu, in vBNST. At 48 h after naloxone-induced morphine withdrawal there was a further significant increase in the extracellular levels of NE, but not Glu, in vBNST. The presence of UK 14304, an ,2 -adrenergic agonist, induced a significant decrease in NE extracellular levels in all experimental groups. In contrast, UK 14304 induced a significant decrease in Glu extracellular levels only in saline-treated rats. The results also show that the vBNST presents a rostrocaudal gradient of NE and contains 9.4% of total brain NE. The increase in NE extracellular levels in vBNST induced by chronic morphine treatment and the further increase in NE levels 48 h after naloxone-induced morphine withdrawal suggest that NE in vBNST may be involved in the pharmacological effects of chronic morphine and withdrawal. [source]

Growth factors and proliferation of cultured rat gingival cells in response to cyclosporin A

JOURNAL OF PERIODONTAL RESEARCH, Issue 1 2005
Takumasa Yoshida
Objective:, The prominent side-effect of cyclosporin A, an immunosuppressive drug, in oral tissues is gingival outgrowth, although the exact mechanism underlying this side-effect is unclear. The main purposes of the present study were to determine whether cyclosporin A induced the gingival outgrowth by promoting proliferation of gingival cells and whether growth factors such as transforming growth factor-,s (TGF-,s), fibroblast growth factor-2 (FGF-2), platelet-derived growth factors (PDGFs), and insulin-like growth factors (IGFs) are involved in the possible changes in the proliferation of gingival cells induced by cyclosporin A. Methods:, Cells isolated from rat gingival tissues were cultured with cyclosporin A or IGF-I for 3 days. The effects of cyclosporin A or IGF-I on the proliferation of cultured rat gingival cells were analyzed with a CellTiter 96 proliferation assay kit. The mRNA expression levels for TGF-,s, FGF-2, PDGFs, IGFs, insulin-like growth factor receptors (IGFRs), and insulin-like growth factor binding proteins (IGFBPs) in the rat gingival cells treated with cyclosporin A were measured using competitive reverse transcription,polymerase chain reaction (RT,PCR). Results:, Cyclosporin A induced 23,25% (p < 0.001) increases in the proliferation of rat gingival cells and approximately 130% (p < 0.05) and 60% (p < 0.05) elevations in the mRNA expression levels for TGF-,1 and FGF-2, respectively. On the other hand, exogenous IGF-I induced 8,11% (p < 0.05) increases in the proliferation, but cyclosporin A induced 30,80% (p < 0.05,0.01) reductions in the mRNA expression levels for endogenous IGF-I, IGFR1, IGFBP2, IGFBP3, IGFBP5, and IGFBP6. Conclusions:, Cyclosporin A stimulates the proliferation of rat gingival cells. TGF-,1 and FGF-2 could be involved, but IGFs, IGFRs and IGFBPs could not be directly involved in this cyclosporin A induced-stimulation of the gingival cell proliferation. [source]

Induction of apoptosis of RAW 264.7 cells by the cytostatic macrolide apicularen A

JOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 9 2003
JangJa Hong
ABSTRACT In RAW 264.7 cells, a mouse leukaemic monocyte cell line, apicularen A decreased cell growth and survival as assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in a concentration-dependent manner at 10,1000 nM. Apicularen B, an N -acetyl-glucosamine glycoside of apicularen A, was 10,100-fold less effective than apicularen A. Apicularen A induced a DNA ladder, an increase in the percentage of sub-G1 cells and annexin V-binding cells, and promoted the activation of caspase as revealed by the cleavage of poly(ADP-ribose) polymerase, indicating that apicularen A induced apoptosis in RAW 264.7 cells. In addition, apicularen A phosphorylated p44/42 mitogen-activated protein kinase (MAPK) and p38 MAPK. The p44/42 MAPK inhibitor PD98059 rescued the cells from apicularen-induced decrease in cell growth and survival as determined by the MTT assay, while the p38 MAPK inhibitor SB203580 augmented the effect of apicularen A. This suggested the activation of p44/42 MAPK to be pro-apoptotic and the activation of p38 MAPK anti-apoptotic in apicularen A-treated RAW 264.7 cells. [source]

Polymerization of substituted phenylacetylenes with a novel, water-soluble Rh,vinyl complex in water

JOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 9 2004
Keiji Kanki
Abstract A novel, water-soluble Rh complex, (nbd)Rh[PPh2(m -NaOSO2C6H4)] [C(Ph)CPh2] (1) was synthesized by the reaction of [(nbd)RhCl]2, Ph2P(m -NaOSO2C6H4) and Ph2CC(Ph)Li, whose structure was determined by NMR and IR spectroscopies. The Rh catalyst 1 induced the polymerization of phenylacetylene (PA) in water to give two kinds of polymers; one was soluble in organic solvents such as tetrahydrofuran (THF) and CHCl3, and the other was insoluble in common organic solvents. The polymerization of sodium p -ethynylbenzoate (p -NaOCO-PA) homogeneously proceeded with 1 in water at 60 °C to give the polymer in high yield. Poly(p -NaOCO-PA) was treated with 1 N HCl and then reacted with (CH3)3SiCHN2 to obtain poly(p -MeOCO-PA). The methyl-esterified polymer was insoluble in THF and CHCl3, which suggests that the formed poly(p -MeOCO-PA) has cis,cisoidal structure. The polymer obtained from the polymerization of [p -CH3(OCH2CH2)2O2CC6H4]CCH with 1 in water was soluble in methanol, ethanol, and THF, and partly soluble in water. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 2100,2105, 2004 [source]

Plasminogen activator inhibitor-1 contributes to the deleterious effect of obesity on the outcome of thrombotic ischemic stroke in mice

JOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 8 2007
N. NAGAI
Summary.,Background:,It is widely accepted that obesity is a risk factor for ischemic heart disease, but the association with stroke is less clear. Adipose tissue is an important source of plasminogen activator inhibitor-1 (PAI-1), the main inhibitor of plasminogen activation. Objective:,To test the hypothesis that elevated PAI-1 levels associated with obesity negatively affect the outcome of thrombotic ischemic stroke. Methods:,Middle cerebral artery (MCA) occlusion was induced photochemically in mice with nutritionally induced or genetically determined obesity and their lean counterparts. Results:,The MCA occlusion time (to obtain complete occlusion) was significantly shorter in obese (nutritionally induced) than in lean wild-type (WT) C57Bl/6 mice, whereas the infarct size was significantly larger and intracranial hemorrhage (ICH) was enhanced (all P < 0.05). Similar observations were made in genetically obese ob/ob mice, as compared to lean WT littermates. In both strains, obesity was associated with markedly elevated circulating PAI-1 levels, probably originating from the fat tissue. In contrast, PAI-1-deficient lean and obese mice did not display significant differences in MCA occlusion time, infarct volume or ICH. Conclusions:,Plasminogen activator inhibitor-1 may play a functional role in the deleterious effect of obesity on the outcome of thrombotic ischemic stroke in mice. [source]

Alterations of intestinal motor responses to various stimuli after Nippostrongylus brasiliensis infection in rats: role of mast cells

NEUROGASTROENTEROLOGY & MOTILITY, Issue 3 2000
J. Gay
Nippostrongylus brasiliensis infection induces jejunal mastocytosis associated with enteric nerve remodelling in rats. The aim of this study was to evaluate the intestinal motility responses to meals and to neurotransmitters involved in the control of gut motility (acetylcholine (carbachol), substance P and neurokinin A) in both control and N. brasiliensis -infected rats 30 days post-infection. All rats were equipped with NiCr electrodes in the jejunum to record myoelectrical activity. The duration of disruption of the jejunal migrating myoelectrical complexes (MMC) induced by the different stimuli was determined. Meal ingestion and substance P administration disrupted the MMC pattern for similar durations in the two groups. Carbachol and neurokinin A induced a significantly longer MMC disruption in post-infected rats than in controls (125 ± 8.3 vs. 70 ± 6 min for carbachol 100 ,g kg,1 and 51 ± 4 vs. 40 ± 2 for neurokinin A 50 ,g kg,1). The enhanced motor response in postinfected rats was reduced by previous mast cell stabilization with ketotifen or mast cell degranulation with compound BrX 537 A. In conclusion, the increased intestinal motor reactivity to carbachol and neurokinin A in post- N. brasiliensis -infected rats depends upon intestinal mast cell hyperplasia and degranulation. [source]

, -secretase inhibitors exerts antitumor activity via down-regulation of Notch and Nuclear factor kappa B in human tongue carcinoma cells

ORAL DISEASES, Issue 6 2007
J Yao
Objective:, To investigate the effect of the , -secretase inhibitors (GSIs) on the growth of human tongue carcinoma cells and to provide the molecular mechanism for potential application of GSIs in the treatment of tongue carcinoma. Materials and methods:, Human tongue carcinoma Tca8113 cells were cultured with the GSI L-685 458. Cell growth was determined by the methylthiazole tetrazolium method. Cell cycle and apoptosis were analyzed by flow cytometry and/or confocal microscopy. RT-PCR and Western blot were employed to determine the intracellular expression levels. Nuclear factor kappa B (NF- ,B) activation was examined by electrophoretic mobility shift assay. Results:, L-685,458 dose-dependently inhibited the growth of human tongue carcinoma Tca8113 cells by inducing G0,G1 cell cycle arrest and apoptosis. The mRNA and protein levels of Hairy/Enhancer of Split-1, a target of Notch activation, were decreased dose-dependently by L-685,458. Furthermore, L-685,458 down-regulated cyclin D1, B-cell lymphocytic-leukemia proto-oncogene 2 and c-Myc expressions, which are regulated by the transcription factor NF- ,B. Coincident with this observation, L-685,458 induced a dose-dependent reduction of constitutive NF- ,B activation in Tca8113 cells. Conclusions:, The GSI L-685,458 may have a therapeutic value for the treatment of human tongue carcinoma. Moreover, the effects of L-685,458 in tumor inhibition may act partially via the modulation of Notch and NF- ,B. [source]

Mapping of Atrial Activation Patterns After Inducing Contiguous Radiofrequency Lesions: An Experimental Study

PACING AND CLINICAL ELECTROPHYSIOLOGY, Issue 2 2001
FRANCISCO J. CHORRO
CHORRO, F.J., et al.: Mapping of Atrial Activation Patterns After Inducing Contiguous Radiofrequency Lesions: An Experimental Study. High resolution mapping techniques are used to analyze the changes in atrial activation patterns produced by contiguous RF induced lesions. In 12 Langendorff-perfused rabbit hearts, left atrial activation maps were obtained before and after RF induction of epicardial lesions following a triple-phase sequential protocol: (phase 1) three separate lesions positioned vertically in the central zone of the left atrial wall; (phase 2) the addition of two lesions located between the central lesion and the upper and lower lesions; and (phase 3) the placement of four additional lesions between those induced in the previous phases. In six additional experiments a pathological analysis of the individual RF lesions was performed. In phase 1 (lesion diameter = 2.8 ± 0.2 mm, gap between lesions = 3 ± 0.8 mm), the activation process bordered the lesions line in two (2.0-ms cycles) and four experiments (1.0-ms cycles). In phase 2, activation bordered the lesions line in eight (2.0-ms cycles, P < 0.01 vs control) and nine experiments (1.0-ms cycles, P < 0.001), and in phase 3 this occurred in all experiments except one (both cycles, P < 0.001 vs control). In the experiments with conduction block, the increment of the interval between activation times proximal and distal to the lesions showed a significant correlation to the length of the lesions (r = 0.68, P < 0.05, 100-ms cycle). In two (17%) experiments, sustained regular tachycardias were induced with reentrant activation patterns around the lesions line. In conclusion, in this acute model, atrial RF lesions with intact tissue gaps of 3 mm between them interrupt conduction occasionally, and conduction block may be frequency dependent. Lesion overlap is required to achieve complete conduction block lines. Tachycardias with reentrant activation patterns around a lesions line may be induced. [source]

Cytokine induction by respiratory syncytial virus and adenovirus in bronchial epithelial cells

PEDIATRIC PULMONOLOGY, Issue 3 2007
Jong-Seo Yoon MD
Abstract In order to broaden our knowledge of the primary immune responses to respiratory syncytial virus (RSV) and adenovirus infections, we compared the concentrations of interleukin (IL)-6, IL-8, and regulated on activation, normal T cell expressed and secreted (RANTES) produced in vitro during RSV and adenovirus infections of bronchial epithelial cells. We infected BEAS-2B cells,a human bronchial epithelial cell line,with RSV, adenovirus serotype 3, or serotype 7 and measured the concentrations of IL-6, IL-8, and RANTES in the cell culture supernatants. When the multiplicity of infection (MOI) was 1, RSV induced the production of markedly higher concentrations of IL-6, IL-8, and RANTES than the adenovirus. When the MOI of the adenovirus was increased to 100, the production of IL-6 and IL-8 increased. However, the amounts produced were still lower than those produced by RSV with the MOI of 1. There was no statistically significant increase in the production of RANTES in spite of the MOI of the adenovirus was increased to 100. Adenovirus serotype 7 induced the production of considerably more IL-6 and IL-8 than serotype 3 in the MOI of 100. However, neither adenovirus serotype triggered an increase in the production of RANTES in spite of the MOI of 100. This demonstrates that RSV could have a superior capacity to stimulate the production of IL-6, IL-8, and RANTES in the bronchial epithelial cells. This study may help to explain the differences in the clinical outcomes of RSV and adenovirus infections. Pediatr Pulmonol. 2007; 42:277,282. © 2007 Wiley-Liss, Inc. [source]

Induction of Persistent Double Strand Breaks Following Multiphoton Irradiation of Cycling and G1 -arrested Mammalian Cells,Replication-induced Double Strand Breaks

PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 6 2008
Jane V. Harper
DNA double strand breaks (DSBs) are amongst the most deleterious lesions induced within the cell following exposure to ionizing radiation. Mammalian cells repair these breaks predominantly via the nonhomologous end joining pathway which is active throughout the cell cycle and is error prone. The alternative pathway for repair of DSBs is homologous recombination (HR) which is error free and active during S- and G2/M-phases of the cell cycle. We have utilized near-infrared laser radiation to induce DNA damage in individual mammalian cells through multiphoton excitation processes to investigate the dynamics of single cell DNA damage processing. We have used immunofluorescent imaging of ,-H2AX (a marker for DSBs) in mammalian cells and investigated the colocalization of this protein with ATM, p53 binding protein 1 and RAD51, an integral protein of the HR DNA repair pathway. We have observed persistent DSBs at later times postlaser irradiation which are indicative of DSBs arising at replication, presumably from UV photoproducts or clustered damage containing single strand breaks. Cell cycle studies have shown that in G1 cells, a significant fraction of multiphoton laser-induced prompt DSBs persists for >4 h in addition to those induced at replication. [source]

New 5-Aminolevulinic Acid Esters,Efficient Protoporphyrin Precursors for Photodetection and Photodynamic Therapy,

PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 5 2003
H. Brunner
ABSTRACT Photodetection (PD) and photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA),induced protoporphyrin IX (PPIX) accumulation are approaches to detect and treat dysplasia and early cancer in the gastrointestinal tract and in the urinary bladder. Because ALA-induced PPIX production is limited, we synthesized ALA ester hydrochlorides 3,22 and tested them in two different in vitro models (gastrointestinal tract: HT29,CCD18; urinary bladder: J82,UROTSA). PPIX accumulation after incubation with 0.12 mmol/L for 3 h and PPIX accumulation as a function of different incubation times were measured using flow cytometry. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were performed to check cellular dark toxicity. Phototoxicity after irradiation was tested. ALA nonafluorohexylester hydrochloride 11, ALA thiohexylester hydrochloride 13 and ALA dibenzyldiester dihydrochloride 19 induced appreciably increased PPIX levels and showed improved phototoxicity compared with the references ALA hydrochloride 1, ALA hexylester hydrochloride 3 and ALA benzylester hydrochloride 4. Thus, the new compounds 11, 13 and 19 are promising compounds for PD and PDT. [source]