bluuebunny/crossref_metadata_2025_split huggingface dataset:
Postnatal development of synaptopodin expression in the rodent hippocampus
Authors: Czarnecki Kathrin , Haas Carola A. , Bas Orth Carlos , Deller Thomas , Frotscher Michael 
Published: July 2005

AbstractSynaptopodin is an actin‐binding protein of renal podocytes and dendritic spines. We have recently shown that synaptopodin is localized to the spine apparatus, a characteristic organelle of dendritic spines on forebrain neurons. Synaptopodin‐deficient mice do not form spine apparatuses, indicating a role of synaptopodin in the formation of this organelle. Here we studied the development of synaptopodin expression in the postnatal rat hippocampus. At birth, synaptopodin mRNA is mainly expressed in CA3 pyramidal neurons. At postnatal day (P) 6, synaptopodin mRNA expression is still strongest in CA3 but is now also found in CA1 pyramidal neurons and granule cells of the suprapyramidal blade of the dentate gyrus. At P9, an almost adult pattern is seen with synaptopodin mRNA expressed by virtually all principal neurons. While synaptopodin mRNA was restricted to cell somata, immunostaining for synaptopodin protein labeled dendritic layers. At birth, no immunoreactivity was visible, while at P5 a weak staining mainly in stratum oriens was observed. At P9, immunolabeling was still strongest in stratum oriens followed by the molecular layer of the dentate gyrus. The adult pattern with strong labeling of all dendritic layers was reached by P12. Together these findings show that synaptopodin expression follows the well‐known sequence of hippocampal principal neuron development. Unexpectedly, we also observed synaptopodin mRNA expression in a small population of interneurons as revealed by double labeling with interneuron markers. However, no immunolabeling for synaptopodin was observed in identified interneurons, confirming that the protein is mainly present in spine‐bearing principal cells. J. Comp. Neurol. 490:133–144, 2005. © 2005 Wiley‐Liss, Inc.
Insular and gustatory inputs to the caudal ventral striatum in primates
Authors: Fudge Julie L. , Breitbart Michael A. , Danish Matthew , Pannoni Valerie 
Published: July 2005

AbstractThe ventral striatum mediates goal‐directed behaviors based, in part, on inputs from the amygdala. However, striatal areas caudal to the ventral striatum also receive inputs from the amygdala. In primates, the amygdala projects to the central ventral putamen, lateral amygdalostriatal area, and caudal ventral putamen, suggesting that these regions are also “limbic‐related.” The anterior insula, which integrates sensory and amygdaloid inputs, projects to the classic ventral striatum. We used retrograde and anterograde tract tracing techniques to determine the extent to which specific subdivisions of the insula influence the caudal ventral striatum in the primate. The anterior (agranular and rostral dysgranular) insula has significant inputs to caudal ventral striatal regions that receive projections from the amygdala. In contrast, the posterior (granular) insula has sparse projections. Within the agranular insula, the posteromedial agranular (Iapm), lateral agranular (Ial), and posterolateral agranular (Iapl) subdivisions have the strongest inputs. These subdivisions mediate olfactory, gustatory, and visceral information processing (Carmichael and Price JL [1996b] J. Comp. Neurol. 363:642–640). In contrast, the intermediate agranular subdivision (Iai) is relatively devoid of visceral/gustatory inputs and has few inputs. In summary, caudal ventral striatal areas that receive amygdaloid inputs also receive significant innervation by agranular and dysgranular insula subdivisions that are themselves connected with the amygdala. Within this projection, the Ial, Iapm, and Iapl make the strongest contribution, suggesting that highly processed visceral/autonomic information, taste, and olfaction influence behavioral responses mediated by the caudal ventral striatum. J. Comp. Neurol. 490:101–118, 2005. © 2005 Wiley‐Liss, Inc.
Engineering micropatterned surfaces for the coculture of hepatocytes and Kupffer cells
Authors: Zinchenko Yekaterina S. , Coger Robin N. 
Published: July 2005

AbstractBioartificial liver (BAL) devices are used for applications ranging from pharmaceutical testing to temporary liver replacement. The capabilities of these devices can be improved by optimizing the range of hepatocyte functions that the BAL is able to perform. One means of achieving this is to design the BAL such that it establishes communication between hepatocytes and nonparenchymal cells. To understand how these heterotypic interactions can be favorably utilized in BAL design, it is first necessary to establish a culture environment that permits the controlled interactions of multiple cell types. This is the goal of the current study, which focuses on micropatterned cocultures of hepatocytes with Kupffer cells. The micropatterning technique relies on a polydimethylsiloxane (PDMS) membrane to achieve various two‐dimensional configurations of the ECM prior to seeding the cell populations. The easy and inexpensive method of making the PDMS membranes differs from that reported in the literature and is detailed in the current study. To demonstrate the success of the method, surface characterization of the resultant micropatterns, as well as morphological and functional results are also presented. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005
Influence of channel width on alignment of smooth muscle cells by high‐aspect‐ratio microfabricated elastomeric cell culture scaffolds
Authors: Glawe John D. , Hill Jason B. , Mills David K. , McShane Michael J. 
Published: July 2005

AbstractEngineered smooth muscle tissue requires ordered configurations of cells to reproduce native function, and microtechnology offers possibilities for physically and chemically controlling cell organization with high spatial resolution. In this work, poly(dimethylsiloxane) microchannel scaffolds, modified by layer‐by‐layer self‐assembly of polyelectrolytes to promote cell adhesion, were evaluated for use as substrates for the culture of aligned smooth muscle cells. The hypothesis that narrower channels would result in better alignment was tested using channel width dimensions of 20, 30, 40, 50, and 60 μm, in addition to flat (control) surfaces. Alignment of cells was assessed by two different methods, each sensitive to a different aspect of cell alignment from fluorescence micrographs. Two‐dimensional fast Fourier transform analysis was performed to analyze the orientation distribution of actin filaments in cells. This was complemented by connectivity analysis of stained nuclei to obtain nuclear orientation distributions. Both methods produced consistent data that support the hypothesis that narrow microchannels promote a highly aligned culture of smooth muscle cells, and the degree of alignment is dependent on the microchannel width. Precise replication of in vivo cell alignment in engineered tissue, with the ability to tailor specific surface chemistries of the scaffold to the desired application, will potentially allow the production of artificial tissue that more closely duplicates the structure and function of native tissue. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005
Midazolam suppresses thrombin‐induced heat shock protein 27 phosphorylation through inhibition of p38 mitogen‐activated protein kinase in cardiac myocytes
Authors: Tanabe Kumiko , Akamatsu Shigeru , Suga Hidetaka , Takai Shinji , Kato Kanefusa , Dohi Shuji , Kozawa Osamu 
Published: July 2005

AbstractIt has been shown that anesthetics have effects of cardiac preconditioning. Heat shock proteins (HSPs) function as molecular chaperone. Among them, HSP27, a low‐molecular‐weight HSP, abundantly exist in heart. However, the relationship between anesthetics and HSP27 in heart is not yet clarified. We investigated whether thrombin induces or phosphorylates HSP27 in primary cultured mouse myocytes and the effect of midazolam on the thrombin‐stimulated HSP27 phosphorylation and the mechanism behind it. Thrombin time dependently phosphorylated HSP27 at Ser‐15 and Ser‐85 while having no effect on the levels of HSP27. Midazolam markedly suppressed the thrombin‐induced phosphorylation of HSP27 at both Ser‐15 and Ser‐85. Thrombin induced the phosphorylation of p44/p42 MAP kinase and p38 MAP kinase without affecting stress‐activated protein kinase/c‐Jun N‐terminal kinase. In addition, midazolam attenuated the phosphorylation of thrombin‐induced p38 MAP kinase but not that of p44/p42 MAP kinase. SB203580 and PD169316, inhibitors of p38 MAP kinase, suppressed the thrombin‐induced phosphorylation of HSP27 at both Ser‐15 and Ser‐85. These results strongly suggest that thrombin induces the HSP27 phosphorylation at least through the p38 MAP kinase activation in cardiac myocytes and that midazolam inhibits the thrombin‐induced HSP27 phosphorylation via suppression of p38 MAP kinase activation. © 2005 Wiley‐Liss, Inc.
Mechanical loading stimulates ecto‐ATPase activity in human tendon cells
Authors: Tsuzaki M. , Bynum D. , Almekinders L. , Faber J. , Banes A.J. 
Published: July 2005

AbstractResponse to external stimuli such as mechanical signals is critical for normal function of cells, especially when subjected to repetitive motion. Tenocytes receive mechanical stimuli from the load‐bearing matrix as tension, compression, and shear stress during tendon gliding. Overloading a tendon by high strain, shear, or repetitive motion can cause matrix damage. Injury may induce cytokine expression, matrix metalloproteinase (MMP) expression and activation resulting in loss of biomechanical properties. These changes may result in tendinosis or tendinopathy. Alternatively, an immediate effector molecule may exist that acts in a signal‐dampening pathway. Adenosine 5′‐triphosphate (ATP) is a candidate signal blocker of mechanical stimuli. ATP suppresses load‐inducible inflammatory genes in human tendon cells in vitro. ATP and other extracellular nucleotide signaling are regulated efficiently by two distinct mechanisms: purinoceptors via specific receptor–ligand binding and ecto‐nucleotidases via the hydrolysis of specific nucleotide substrates. ATP is released from tendon cells by mechanical loading or by uridine 5′‐triphosphate (UTP) stimulation. We hypothesized that mechanical loading might stimulate ecto‐ATPase activity. Human tendon cells of surface epitenon (TSC) and internal compartment (TIF) were cyclically stretched (1 Hz, 0.035 strain, 2 h) with or without ATP. Aliquots of the supernatant fluids were collected at various time points, and ATP concentration (ATP) was determined by a luciferin‐luciferase bioluminescence assay. Total RNA was isolated from TSC and TIF (three patients) and mRNA expression for ecto‐nucleotidase was analyzed by RT‐PCR. Human tendon cells secreted ATP in vitro (0.5–1 nM). Exogenous ATP was hydrolyzed within minutes. Mechanical load stimulated ATPase activity. ATP was hydrolyzed in mechanically loaded cultures at a significantly greater rate compared to no load controls. Tenocytes (TSC and TIF) expressed ecto‐nucleotidase mRNA (ENTPD3 and ENPP1, ENPP2). These data suggest that motion may release ATP from tendon cells in vivo, where ecto‐ATPase may also be activated to hydrolyze ATP quickly. Ecto‐ATPase may act as a co‐modulator in ATP load‐signal modulation by regulating the half‐life of extracellular purine nucleotides. The extracellular ATP/ATPase system may be important for tendon homeostasis by protecting tendon cells from responding to excessive load signals and activating injurious pathways. © 2005 Wiley‐Liss, Inc.
Roles of Ca2+ and the Ca2+‐sensing receptor (CASR) in the expression of inducible NOS (nitric oxide synthase)‐2 and its BH4 (tetrahydrobiopterin)‐dependent activation in cytokine‐stimulated adult human astrocytes
Authors: Dal Pra Ilaria , Chiarini Anna , Nemeth Edward F. , Armato Ubaldo , Whitfield James F. 
Published: July 2005

AbstractSince NO production by NOS‐2 made by astrocytes activated by proinflammatory cytokines contributes to the killing of neurons in variously damaged human brains, knowing the mechanisms responsible for NOS‐2 expression should contribute to developing effective therapeutics. The expression and activation of NOS‐2 in normal adult human cerebral cortical astrocytes treated with three proinflammatory cytokines, IL‐1β, TNF‐α, and IFN‐γ, are driven by two separable mechanisms. NOS‐2 expression requires a burst of p38 MAPK activity, while the activation of the resulting enzyme protein requires MEK/ERK‐dependent BH4 (tetrahydrobiopterin) synthesis between 24 and 24.5 h after adding the cytokines to the culture medium. Here we show that NOS‐2 expression in the activated astrocytes requires that the culture medium contain 1.8 mM Ca2+, but it is unaffected by inhibiting calcium‐sensing receptors (CASRs) with NPS 89636. However, NOS‐2 activation is inhibited by NPS 89626 during the MEK/ERK‐dependent stage between 24 and 24.5 h after adding the cytokines, and this inhibition can be overridden by exogenous BH4. Therefore, NOS‐2 expression and the subsequent BH4‐dependent NOS‐2‐activation in human astrocytes need 1.8 mM Ca2+ to be in the culture medium, while NOS‐2 activation also needs functional CASRs between 24 and 24.5 h after cytokine addition. These findings raise the possibility that calcilytic drugs prevent NO‐induced damage and death of human neurons. © 2005 Wiley‐Liss, Inc.
Dynamics of DNA repair suggested by the subcellular localization of Brca1 and Brca2 proteins
Authors: Huber L. Julie , Chodosh Lewis A. 
Published: July 2005

AbstractThe localization of proteins to specific subcellular compartments often reveals clues regarding their biological functions. Although significant progress has been made towards understanding how damaged DNA is repaired, experiments to date have primarily focused on signal transduction pathways that activate DNA repair protein complexes and on how these complexes are assembled. Current evidence suggests that certain DNA repair processes are spatially organized such that aberrant DNA structures can be brought into proximity with DNA repair proteins at fixed sites. Since biochemical evidence suggests that the tumor suppressor proteins, Brca1 and Brca2, may mediate the assembly of protein complexes involved in the repair of damaged DNA, we have performed subcellular fractionation experiments to determine the subnuclear localization of these proteins. The majority of Brca1 and Brca2 proteins were found to interact tightly with the nuclear matrix. Furthermore, within the limits of detection, localization of Brca1 and Brca2 to the nuclear matrix was not altered following treatment of cells with DNA damaging agents that activate homology‐mediated double‐stranded DNA break and transcription‐coupled repair pathways. Our findings suggest that Brca1 and Brca2 may perform their DNA repair‐related functions from positions that are anchored to the nuclear matrix. These data are consistent with proposed models that suggest that components of specific repair complexes residing on the nuclear matrix function to recruit damaged DNA. © 2005 Wiley‐Liss, Inc.
PAR1‐mediated NFκB activation promotes survival of prostate cancer cells through a Bcl‐xL‐dependent mechanism
Authors: Tantivejkul Kwanchanit , Loberg Robert D. , Mawocha Samkeliso C. , Day Lashon L. , John Lauren St. , Pienta Brian A. , Rubin Mark A. , Pienta Kenneth J. 
Published: July 2005

AbstractWe have previously reported that protease‐activated receptor 1 (PAR1 or thrombin receptor) is over‐expressed in metastatic prostate cancer cell lines compared to prostate epithelial cells. In this study, we examined 1,074 prostate biopsies by tissue microarray analysis and demonstrated that PAR1 expression is significantly increased in prostate cancer compared to normal prostate epithelial cells and benign prostatic hyperplasia. We hypothesized that PAR1 activation contributed to prostate cancer cell progression. We demonstrated that stimulation of PAR1 by thrombin or thrombin receptor activating peptide (TRAP6), in androgen‐independent DU145 and PC‐3 cells resulted in increased DNA binding activity of the NFκB p65 subunit. IL‐6 and IL‐8 levels were also elevated in conditioned media by at least two‐fold within 4–6 h of PAR1 activation. This induction of cytokine production was abrogated by pretreatment of cells with the NFκB inhibitor caffeic acid phorbol ester. The p38 and ERK1/2 MAPK signaling cascades were also activated by PAR1 stimulation, whereas the SAPK/JNK pathway was unaffected. Inhibition of p38 and ERK1/2 by SB‐203589 and PD‐098059, respectively, did not abrogate NFκB activity, suggesting an independent induction of NFκB by PAR1 stimulation. Furthermore, TUNEL assay showed that activation of PAR1 attenuated docetaxel induced apoptosis through the upregulation of the Bcl‐2 family protein Bcl‐xL. Akt activation was not observed, suggesting that drug resistance induced by PAR1 was independent of PI3K signaling pathway. Because thrombin and PAR1 are over‐expressed in prostate cancer patients, targeting the inhibition of their interaction may attenuate NFκB signaling transduction resulting in decreased drug resistance and subsequent survival of prostate cancer cells. © 2005 Wiley‐Liss, Inc.
Recombinant polyclonal antibodies for cancer therapy
Authors: Sharon Jacqueline , Liebman Meredith A. , Williams Brent R. 
Published: July 2005

AbstractAlthough monoclonal antibodies are increasingly used for cancer therapy, remissions are only temporary due to emergence of tumor cell escape variants that are no longer affected by the antibody. The emergence of escape variants could be minimized by multi‐targeting of tumor cells with polyclonal antibodies, which would also be more efficient than monoclonal antibodies at mediating effector functions for target destruction. A technology for generating recombinant polyclonal antibodies for cancer therapy has been developed based on the construction and selection of tumor‐reactive Fab phage display libraries. The selected Fabs are mass‐converted to full‐length polyclonal antibody libraries (PCALs) of any isotype and any species. Prototypic PCALs generated against human colorectal cancer cell lines showed that libraries of diverse recombinant antibodies, enriched for reactivity to the cancer cells compared to normal human cells, can be obtained. The success of recombinant polyclonal antibodies as cancer therapeutics will depend on the ability to generate, characterize, and mass‐produce PCALs with high ratios of cancer‐to‐normal reactivities that cross‐react with many cancers of the same type. © 2005 Wiley‐Liss, Inc.
Garcinol modulates tyrosine phosphorylation of FAK and subsequently induces apoptosis through down‐regulation of Src, ERK, and Akt survival signaling in human colon cancer cells
Authors: Liao Chiung‐Ho , Sang Shengmin , Ho Chi‐Tang , Lin Jen‐Kun 
Published: July 2005

AbstractGarcinol, from the fruit rind of Garcinia indica and other species, has been reported to suppress colonic aberrant crypt foci (ACF) formation in rats. In this study, we investigate the beneficial effects of tumor prevention by garcinol on the human colorectal cancer cell line, HT‐29. Focal adhesion kinase (FAK) is the major signaling mediator of integrin‐mediated cell–matrix contact‐regulated cellular proliferation, migration, and apoptosis in adherent cells. Results of Matrigel analysis show that exposure of HT‐29 cells to 10 µM garcinol inhibited cell invasion, and decreased the dose‐dependent tyrosine phosphorylation of FAK. We further demonstrate by Western blot analysis that garcinol inhibited activation of the Src, MAPK/ERK, and PI3K/Akt signaling pathways. To investigate whether the loss of integrin‐mediated cell–matrix contact can induce apoptosis, we demonstrate that garcinol induced it in HT‐29 cells. The apoptotic dose of garcinol (20 µM) changed the ratio of the anti‐apoptotic Bcl‐2 and proapoptotic BAX proteins within 12 h, which correlated with a release of cytochrome c from the mitochondria to the cytosol, and with PARP cleavage. Additionally, we demonstrate that a decreasing MMP‐7 protein level in HT‐29 cells results in sensitization to garcinol. Garcinol also significantly inhibited the expression of MMP‐7 in IL‐1β‐induced HT‐29 cells. These results suggest that garcinol reduces cell invasion and survival through the inhibition of FAK's downstream signaling. © 2005 Wiley‐Liss, Inc.
Iron–ascorbic acid‐induced oxidant stress and its quenching by paraoxonase 1 in HDL and the liver: Comparison between humans and rats
Authors: Trudel K. , Sinnett D. , James R.W. , Delvin E. , Amre D. , Seidman E. , Levy E. 
Published: July 2005

AbstractParaoxonase 1 (PON1) is a serum enzyme closely associated with high‐density lipoprotein (HDL), which may protect against atherosclerosis by hydrolyzing lipid peroxides and several organophosphorus compounds. The purpose of the present study was to test the hypothesis that lipid peroxidation modifies the activity and protein mass of PON1 in humans and rats. Our findings revealed that the bulk of the activity monitored by the hydrolysis of paraoxon and phenyl acetate was confined to liver intracellular endoplasmic reticulum‐derived microsomes and was mostly recovered in circulating HDL3. Confirmation was obtained by the determination of PON1 expression by Western blot. It is noteworthy that PON1 levels were consistently decreased in human sera, HDL, and liver microsomes compared with rat counterparts. Concomitant with iron‐ascorbate‐mediated lipid peroxidation, there was a decline in PON1 activity and protein in both HDL3 and microsomes, which was attenuated by butylated hydroxytoluene antioxidant treatment. The current data indicate that PON1 localization in microsomes and HDL3 could represent a selective cellular and lipoprotein response to oxidative stress. This was tested by the iron‐ascorbate oxygen‐radical generating system. It is also proposed that the increased PON1 level may have a function related to the well‐known atherosclerosis resistance of rats. © 2005 Wiley‐Liss, Inc.
Salvianolic acid B attenuates plasminogen activator inhibitor type 1 production in TNF‐α treated human umbilical vein endothelial cells
Authors: Zhou Zhe , Liu Yong , Miao Ai‐Dong , Wang Sheng‐Qi 
Published: July 2005

AbstractPlasminogen activator inhibitor type 1 (PAI‐1), which plays a role in the development of atherosclerosis, is produced by endothelial cells following stimulation with various inflammatory cytokines such as tumor necrosis factor (TNF‐α). In the present study, we investigated the effects of a potent water‐soluble antioxidant, salvianolic acid B (SalB; derived from the Chinese herb, Salvia miltiorrhiza), on the expression of PAI‐1 in TNF‐α‐treated human umbilical vein endothelial cells (HUVECs). We found that SalB inhibited TNF‐α‐induced PAI‐1 mRNA production and protein secretion in HUVECs. Treatment with SalB (0.05 and 0.15 µM) notably attenuated TNF‐α induced expression of PAI‐1 to 90.5% and 74.6%, respectively, after 12 h, and to 75.1% and 64.2%, respectively, after 18 h. We also observed a dose‐dependent decrease in PAI‐1 protein production in the presence of SalB. We then used pathway inhibitors to investigate which step of the TNF‐α induced signaling pathway was targeted by SalB. We found that the c‐Jun N‐terminal kinase (JNK) inhibitor, SP600125, increased the inhibitory effects of SalB on TNF‐α‐induced PAI‐1 secretion, whereas the nuclear factor‐κB (NF‐κB) inhibitor, emodin, and the extracellular signal‐regulated kinase (ERK) inhibitor, PD98059, did not. A gel shift assay further showed that SalB inhibited the TNF‐α‐activated NF‐κB and AP‐1 DNA binding activities in a dose‐dependent manner. Collectively, these results indicate that the NF‐κB and ERK‐AP‐1 pathways are possible targets of SalB in the regulation of TNF‐α‐stimulated PAI‐1 production in HUVECs. © 2005 Wiley‐Liss, Inc.