bluuebunny/crossref_metadata_2025 huggingface dataset:
Retrograde Injections of Formaldehyde Into the Biliary Tree Induce Alterations of Biliary Epithelial Function in Rats
Authors: Dumont M , D'Hont C , Moreau A , Mbape H , Feldmann G , Erlinger S
Published: November 2004

Formaldehyde may induce severe lesions of intrahepatic and extrahepatic bile ducts. The purpose of this study was to examine

in vivo

the functional consequences of an alteration of the biliary epithelium induced by a retrograde intrabiliary injection of formaldehyde in rats. After basal bile collection, a 10% formaldehyde solution was injected into the biliary tree of anesthetized rats, and the cannula was occluded for 30 minutes. Choleresis was then reestablished, and bile flow, bile acid, and bicarbonate secretion were measured both spontaneously and during ursodeoxycholate infusions. Formaldehyde injections induced a significant increase in bile flow and a marked inhibition of ursodeoxycholate–induced increase in biliary bicarbonate concentration and secretion. Biliary glucose secretion, which is normally very low, was increased about 20–fold in animals injected with formaldehyde. Histological and ultrastructural examination of the liver showed alterations of biliary epithelial cells, whereas hepatocytes, bile canaliculi, and canalicular tight junctions remained normal. Hepatocytic excretory function, as assessed by biliary secretion of bile acids, was not affected. It was concluded that short–term formaldehyde intrabiliary injections cause an inhibition of ursodeoxycholate–induced hypersecretion of bicarbonate, an increase in biliary glucose secretion, and selective structural alterations of biliary epithelial cells. These results suggest that formaldehyde retrograde biliary injection may be a useful model to study alterations of biliary epithelial function

in vivo

.
Heat Shock Response Inhibits Cytokine–Inducible Nitric Oxide Synthase Expression in Rat Hepatocytes
Authors: de Vera M E , Kim Y M , Wong H R , Wang Q , Billiar T R , Geller D A
Published: November 2004

During sepsis or inflammation, the liver expresses various protective phenotypes such as the acute phase response or the heat shock response (HSR). Inducible nitric oxide synthase (NOS2) is also expressed in the liver in these conditions and may protect the liver under some circumstances and promote injury in others. We have previously reported that the acute phase response and NOS2 expression are differentially regulated, though both can be expressed simultaneously. The HSR is known to prevent expression of other genes, but its effects on NOS2 expression in the liver is unknown. Therefore, we examined how the HSR influences NOS2 expression in primary rat hepatocytes. Sodium arsenite (Ars) or hyperthermia (43 degrees C) induced the synthesis of hsp72 messenger RNA (mRNA) and protein in hepatocytes, indicating activation of the HSR. In the absence of the HSR, combinations of interleukin– 1β (IL–1β), tumor necrosis factor α (TNF–α), and interferon γ (IFN–γ) stimulated high levels of NOS2 mRNA and nitric oxide (NO) synthesis. However, treatment with Ars or heat shock significantly attenuated cytokine–induced NOS2 mRNA and NO production. The addition of the nuclear factor κB (NF–κB) inhibitor pyrrolidine dithiocarbamate also inhibited NOS2 expression, suggesting a role for NF–κB in the cytokine induction of NOS2 in hepatocytes. Cytokines induced the appearance of an NF–κB complex as shown in gel retardation assays; however, induction of the HSR by Ars partially prevented cytokine–induced formation of this band while hyperthermia had a more complete inhibition. Furthermore, preinduction of the HSR prevented the activation of the NOS2 promoter construct in hepatocytes transfected with a 1.6 kilobase NOS2 promoter linked to luciferase. These findings show that NO production in stressed cells can be modulated by the HSR, possibly through repression of NOS2 gene transcription via the inhibition of NF–κB.
Long–Term Alcohol Intake Enhances Adp–Ribosylation of the Multifunctional Enzyme, Phosphoglucomutase, in Rat Liver
Authors: Nomura F , Noda M , Miyake M , Nakai T
Published: November 2004

Adenosine diphosphate (ADP)–ribosylation is a posttranslational protein modification that, in turn, alters several regulatory proteins in mammalian cells. We demonstrated that long–term alcohol intake enhanced the ADP–ribosylation of a 58–kd protein in rat liver plasma membranes. To assess the biological significance of this phenomenon, we partially purified the 58–kd acceptor protein from solubilized rat liver homogenates by two sequential preparative high–pressure liquid chromatographies. Microsequencing revealed that it was phosphoglucomutase (PGM) (EC 5,4,2,2). This enzyme underwent negligible auto ADP–ribosylation, but the ADP–ribosylation was remarkably increased by adding rat liver plasma membranes. The extent of the increase was greater in alcohol–fed rats than in pair–fed controls, suggesting enhanced enzyme activities toward ADP–ribosylation of PGM after chronic alcohol consumption. Several important enzymes are ADP–ribosylated, after which their activities are modified. The results of this study showed that PGM is a novel substrate for ADP–ribosylation in the liver and that the ADP–ribosylation is increased after chronic alcohol consumption. In view of the variety of roles of PGM in the liver (carbohydrate metabolism and Ca2+ homeostasis), specific roles of this modification in terms of the effects of alcohol on hepatocytes may deserve further investigation.
Tissue–Specific Growth Suppression and Chemosensitivity Promotion in Human Hepatocellular Carcinoma Cells by Retroviral–Mediated Transfer of the Wild–Type P53 Gene
Authors: Xu G W , Sun Z T , Forrester K , Wang X W , Coursen J , Harris C C
Published: November 2004

Selective expression of cytotoxic gene products in tumor cells is one of the goals of gene therapy for treating cancer. We are developing such a strategy for the treatment of human hepatocellular carcinoma (HCC) by linking the wild–type p53 (WT–p53) gene with HCC–associated transcriptional control elements (TCE) to achieve selective growth inhibition of retrovirally transduced HCC cells. Replication–defective, amphotrophic retroviruses were constructed containing a WT–p53 complementary DNA (cDNA) that is transcriptionally regulated by the HCC–associated α–fetoprotein (AFP) gene TCE. Expression of exogenous WT–p53 from this retroviral vector was limited to AFP–producing cells. Introduction of WT–p53 into AFP–positive HCC cells by retroviral infection markedly inhibited their clonal growth in monolayer and soft agar cultures, and increased the sensitivity of these cells to the chemotherapeutic drug, cisplatin. Therefore, restoration of WT–p53 expression in HCC cells, in combination with chemotherapeutic drugs, can be considered as a strategy for the therapy of human liver cancer.
Prophylaxis in Liver Transplant Recipients Using A Fixed Dosing Schedule of Hepatitis B Immunoglobulin
Authors: Terrault N A , Zhou S , Combs C , Hahn J A , Lake J R , Roberts J P , Ascher N L , Wright T L
Published: November 2004

Prophylactic hepatitis B immunoglobulin (HBIg) reduces the risk of reinfection in hepatitis B surface antigen (HBsAg)–positive liver transplant recipients. In the medical center of this study, high–dose HBIg immunoprophylaxis is administered at a fixed dose of 10,000 IU monthly, and in this study, the long–term efficacy of this treatment regimen was examined. Of 52 HBsAg–positive liver transplant recipients, 24 were administered HBIg immunoprophylaxis, and 28 were administered no specific therapy; the 2–year recurrence rates (defined by the reappearance of HBsAg) were 19% and 76%, respectively. Fifty–four percent of the HBIg–treated patients were positive for HBeAg or hepatitis B virus (HBV) DNA (by hybridization assay) pretransplantation. In patients administered monthly HBIg, intrapatient and interpatient variability in trough antibody to HBsAg (anti–HBs) titer was significant, highlighting the potential difficulties of using anti–HBs titer to guide therapy. Trough anti–HBs titers were less in patients who became HBsAg positive than in patients who remained HBsAg–negative (490 vs. 1290 mIU/mL) (

P

= .0001), reflecting either the cause or effect of HBV reinfection. Of 9 patients who remained HBsAg–negative and who were administered monthly HBIg for at least 1 year, HBV DNA by polymerase chain reaction amplification was detectable in the sera of 67%, the lymphocytes of 50%, and the liver of 57%. In conclusion, a fixed monthly dose of HBIg reduces the recurrence of HBs antigenemia, even in patients with indices of active viral replication pretransplantation. The presence of residual virus in the majority of patients administered HBIg suggests that long–term HBIg administration may be necessary.
Viral Dynamics of Hepatitis C Early After Orthotopic Liver Transplantation: Evidence for Rapid Turnover of Serum Virions
Authors: Fukumoto T , Berg T , Ku Y , Bechstein W O , Knoop M , Lemmens H , Lobeck H , Hopf U , Neuhaus P
Published: November 2004

The pathogenesis of hepatitis C virus (HCV) infection is likely to be associated with viral replication in vivo, but little is known concerning the dynamics of HCV turnover. We performed serial measurements of serum HCV–RNA levels following orthotopic liver transplantation (OLT) in nine patients with HCV–positive cirrhosis. Serum HCV–RNA levels were determined by quantitative polymerase chain reaction before, immediately after, and for up to 1 month after OLT. There was a rapid decline in HCV–RNA levels from 3.1 ± 1.3 × 105 copies/ mL (mean ± SEM) preoperatively to 0.15 ± 0.6 × 105 copies/mL on the first and 0.16 ± 0.6 × 105 copies/mL on the second postoperative day (mean viral half–life, 4.0 ± 0.5 h). Thereafter, HCV–RNA levels increased in all but one patient, and by postoperative day 8 reached 3.6 ± 1.3 × 105 copies/mL, exceeding the preoperative levels irrespective of the use or not of rescue immunosuppressive therapy including steroid bolus administration. In most patients, serum virions continued to increase averaging 11.6 ± 2.8 × 105 copies/mL on the 30th postoperative day. These findings indicate that the half– life of HCV is quite short, and that extrahepatic viral replication contributes little to the total virus pool in serum. Furthermore, the marked HCV replication beginning as early as the third postoperative day strongly suggests that the liver graft is rapidly reinfected by HCV after OLT.
The Creatinine Approach to Estimate Skeletal Muscle Mass in Patients With Cirrhosis
Authors: Pirlich M , Selberg O , Boker K , Schwarze M , Muller M J
Published: November 2004

The creatinine–method to estimate muscle mass is frequently used in clinical studies, although the validity of this approach is uncertain in patients with cirrhosis. In this study 102 patients with cirrhosis differing in cause, clinical state, liver, and renal function were investigated to determine whether reduced liver or renal function may explain in part the low levels of urinary creatinine excretion frequently observed in these patients. Muscle mass assessed by 24–hour urinary creatinine excretion was compared with anthropometrically obtained muscle mass calculated from arm muscle area (AMA), and with body cell mass (BCM) estimated by bioelectrical impedance analysis and total body potassium counting. In cirrhosis, the 24–hour urinary creatinine excretion was 10.4% and AMA was 19% lower than predicted values. The differences between the results obtained by different methods did not show any relation to parameters of liver function (ICG– t½, caffeine–t½, MEGX–test, cholinesterase) or the severity of liver disease (i.e., Child–Pugh score). In contrast, renal function was strongly correlated with the differences between creatinine– and anthropometric–muscle mass (r = .64,

P

< .001). At the same time, patients with normal renal function (62% of the whole population) had significantly higher creatinine (29.1 ± 8.5 vs. 15.8 ± 6 kg,

P

< .001) and anthropometric–muscle mass (22.4 ± 6 vs. 17.9 ± 5.3 kg;

P

< .01) than patients with reduced renal function (38% of the patients). In addition, significantly higher differences between measured and predicted values of urinary creatinine excretion (–0.389 ± 0.33 vs. 0.06 ± 0.31 g/24 h;

P

< .001) and of AMA (13.2 ± 12 vs. 7.2 ± 12 cm2;

P

< .03) were found in the subgroup with impaired renal function. In conclusion, renal dysfunction but not reduced liver function systematically affects the urinary creatinine method for the estimation of skeletal muscle mass in cirrhosis.
Research on Marangoni condensation heat transfer for water and ethanol mixture vapor
Authors: Yang‐peng He , Jun‐jie Yan , Yu‐sen Yan , Ji‐ping Liu
Published: November 2004

A brass block was constructed as a test block to study the Marangoni condensation in this paper. The maximal temperature difference of the block surface on which pure steam condensed was 11°C when the block was cooled by the normal temperature water. Regulations and modes of Marangoni condensation for mixture vapor with different mass fractions were studied when the speed of vapor was 0.3 m/s. As both temperature gradients and concentration gradients exist on the condensing surface, the experimental results indicate that the maximal heat transfer coefficient of mixture vapor can be 2.8 times that of pure steam when the Marangoni condensation of mixture vapor appears. The heat transfer coefficient of mixture vapor increases with the decrease of surface subcooling, and it appears a steep increase when the surface subcooling is small enough; the heat transfer flux has a maximum value as the surface subcooling rises; and the different modes of condensation are confirmed when the different ethanol concentration and different surface subcooling exist. © 2004 Wiley Periodicals, Inc. Heat Trans Asian Res, 33(8): 505–514, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/htj.20035
Centrosome abnormalities in ovarian cancer
Authors: Hsu Lih‐Ching , Kapali Malathy , DeLoia Julie A. , Gallion Holly H.
Published: November 2004

Centrosome abnormalities have been found in various cancer types and are thought to be involved in early development of cancer and/or progression. The contribution of centrosome abnormalities to ovarian tumorigenesis has not been previously evaluated. We sought to determine whether centrosome dysfunction occurs in ovarian tumorigenesis, and whether it could be used as an indicator of early neoplastic changes in ovarian surface epithelium (OSE). Primary cultures of normal OSE and ovarian tumors, as well as paraffin‐embedded normal ovaries and ovarian tumors of different stages, were used for immunostaining with a γ‐tubulin antibody. Centrosomes were considered abnormal if there were more than 2 per cell, if their sizes were greater than 2‐fold of a normal centrosome, and/or if they were abnormal in shape. Centrosomes in normal tissue were uniform in size, whereas centrosomes in ovarian tumors tended to be abnormal in size, number and shape. On average, 4.7% of cells in 5 primary normal OSE cultures had more than 2 centrosomes, whereas 14.1% of primary cells from 5 ovarian tumors displayed centrosome abnormalities (p = 0.008). Centrosome abnormalities were present in 60.9% of stage I (n = 23), 83.3% of stage II (n = 30) and all stage III (n = 10) paraffin‐embedded ovarian tumor samples examined, but not in normal tissues. In addition, centrosome abnormalities occurred more frequently in ovarian tumors with higher grade and aggressive serous subtype. This is the first demonstration that centrosome abnormalities occur in ovarian tumors. Centrosome dysfunction may be an early event in ovarian carcinogenesis and involved in ovarian tumor progression. © 2004 Wiley‐Liss, Inc.
Real‐time quantitative PCR determination of urokinase‐type plasminogen activator receptor (uPAR) expression of isolated micrometastatic cells from bone marrow of breast cancer patients
Authors: Pierga Jean‐Yves , Bonneton Charlyne , Magdelénat Henri , Vincent‐Salomon Anne , Nos Claude , Boudou Evelyne , Pouillart Pierre , Thiery Jean‐Paul , de Cremoux Patricia
Published: November 2004

Disseminated tumor cells (DTC) in bone marrow are independently related to poor outcome in patients with breast cancer. Phenotypic characterization of DTC may be useful to improve evaluation of the metastasizing potential of DTC and also to more accurately target aggressive tumor cells. DTC were screened in bone marrow aspirates from breast cancer patients by immunocytochemistry with an anticytokeratin (anti‐CK) antibody (A45B/B3). Because the cell permeabilization and fixation required for intracellular CK staining is deleterious for mRNA, we used microaspiration to isolate single tumor cells stained with a monoclonal antibody directed against a membrane epitope, epithelial cell adhesion molecule (EpCAM), in CK‐positive cases. Urokinase‐type plasminogen activator receptor (uPAR) was quantified by real‐time quantitative RT‐PCR. The SKBR3 human breast cancer cell line was used to calibrate RT‐PCR. A linear relationship was observed between the cycle threshold (Ct) of uPAR and 18S gene expression and SKBR3 cells spiked (1, 3, 7, 10 and 20) in control patient bone marrow. EpCAM‐positive cells were aspirated in 21 out of 25 bone marrow specimens from breast cancer patients with CK‐positive cells and uPAR mRNA expression was determined in 16 cases. A high level of uPAR mRNA in DTC was detected in 8 out of 16 patients (50%) and was associated with a more aggressive primary tumor phenotype (estrogen receptor [ER]‐negative, progesterone receptor [PR]‐negative or HER2‐positive) (p = 0.01). We demonstrated that real‐time quantitative RT‐PCR was reliably adapted to phenotype analysis of isolated micrometastatic cells. A larger study would be useful to confirm the importance of uPAR to define higher risk subgroups of breast cancer patients with micrometastatic disease. © 2004 Wiley‐Liss, Inc.
Development and validation of vertical scanning interferometry as a novel method for acquiring chondrocyte geometry
Authors: Scott C. Corey , Luttge Andreas , Athanasiou Kyriacos A.
Published: November 2004

Chondrocytes are sensitive to changes in shape which depend on the type of substrate, mechanical factors, or biochemical stimuli. Shape changes can cause metabolic and phenotypic alterations. Cell geometry is also important for mechanical models, determination of mechanical properties, and the study of cell attachment and spreading. In this study, a novel method called vertical scanning interferometry (VSI) was developed to allow rapid and straightforward determination of the height, diameter, surface area, volume, and curvature of single chondrocytes. The dimensions of single chondrocytes at 4 and 18 h were obtained and validated. Differences in the heights of zonal chondrocytes were found to be statistically significant. This method was also used to capture the geometry of a cell dehydrating as it was exposed to air. VSI has advantages over confocal microscopy and atomic force microscopy in terms of speed, ease of use, field of view, and precision. VSI quickly obtains and graphically represents the three‐dimensional geometry of chondrocytes in a simple format. These methods could be expanded to image many types of cells on various biomaterials to assess biocompatibility and attachment and cell spreading characteristics. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 72A: 83–90, 2005
Proliferation, activity, and osteogenic differentiation of bone marrow stromal cells cultured on calcium titanium phosphate microspheres
Authors: Barrias Cristina C. , Ribeiro Cristina C. , Lamghari Meriem , Miranda Clara Sá , Barbosa Mário A.
Published: November 2004

In this study, the behavior of bone marrow stromal cells cultured on calcium titanium phosphate (CTP) microspheres was analyzed. Cell adhesion and proliferation were estimated by the neutral red assay and by total DNA quantification. Morphology and deposition of extracellular matrix were assessed by confocal laser scanning microscopy and/or scanning electron microscopy. The expression of the osteoblastic phenotype was evaluated by monitoring alkaline phosphatase activity and osteocalcin secretion. Results revealed that cells were able to attach and spread on the surface of CTP microspheres, and gradually grow into nearly confluent monolayers. Moreover, cells were able to bridge adjacent microspheres forming microsphere–cell clusters. Cells produced an abundant amount of fibrillar extracellular matrix that covered the substrate surface. Alkaline phosphatase activity peaked around days 7–14 and then decreased until day 21. Cells secreted osteocalcin, with higher levels being detected at day 14 than at day 21. Taken together, these results suggest that CTP microspheres are appropriate scaffolds for the growth and differentiation of cells along the osteoblastic lineage. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 72A: 57–66, 2005
Extended release of a novel antidepressant, venlafaxine, based on anionic polyamidoamine dendrimers and poly(ethylene glycol)‐containing semi‐interpenetrating networks
Authors: Yang Hu , Lopina Stephanie T.
Published: November 2004

The multiple daily administration of venlafaxine, a novel third‐generation antidepressant, was reduced based on polyamidoamine and polyethylene glycol (PEG)‐containing semi‐interpenetrating network (IPN), respectively. Venlafaxine was covalently linked to water‐soluble G2.5 anionic polyamidoamine dendrimer via a hydrolyzable ester bond. Semi‐IPN hydrogels were prepared by crosslinking acrylamide in the presence of PEG, and venlafaxine with predetermined amounts was loaded in situ. Dendrimer–venlafaxine conjugate and semi‐IPNs were characterized by proton nuclear magnetic resonance and Fourier transform infrared, respectively. The effect of PEG concentration and molecular weight was studied and discussed for an optimal controlled release. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 72A: 107–114, 2005
Up‐regulation of the association between heat shock protein 90 and endothelial nitric oxide synthase prevents high glucose‐induced apoptosis in human endothelial cells
Authors: Lin Lian‐Yu , Lin Chien‐Yu , Ho Fong‐Ming , Liau Chiau S.
Published: November 2004

Hyperglycemia is the hallmark of diabetes mellitus. Poor glycemic control is correlated with increased cardiovascular morbidity and mortality. High glucose can trigger endothelial cell apoptosis by de‐activation of endothelial nitric oxide synthase (eNOS). eNOS was recently demonstrated to be extensively regulated by Akt and heat shock protein 90 (HSP90). Yet, little is known about the molecular mechanisms that regulate eNOS activity during high glucose exposure. The present study was designed to determine the involvement of protein interactions between eNOS and HSP90 in high glucose‐induced endothelial cell apoptosis. The protein interaction of eNOS/HSP90 and eNOS/Akt were studied in cultured human umbilical vein endothelial cells (HUVECs) exposed to either control‐level (5.5 mM) or high‐level (33 mM) glucose for different durations (2, 4, 6, and 24 h). The results showed that the protein interactions between eNOS and HSP90 and between eNOS and Akt and the phosphorylation of eNOS were up‐regulated by high glucose exposure for 2–4 h. With longer exposures, these effects decreased gradually. During early hours of exposure, the protein interactions of eNOS/HSP90 and eNOS/Akt and the phosphorylation of eNOS were all inhibited by geldanamycin, an HSP90 inhibitor. High glucose‐induced endothelial cell apoptosis was also enhanced by geldanamycin and was reversed by NO donors. LY294002, a phosphatidylinositol 3 (PI3) kinase inhibitor, inhibited the association of eNOS/Akt and the phosphorylation of eNOS but had no effect on the interaction between eNOS and HSP90 during early hours of exposure. From our results we propose that, in HUVECs, during early phase of high glucose exposure, apoptosis can be prevented by enhancement of eNOS activity through augmentation of the protein interaction between eNOS and HSP90 and recruitment of the activated Akt. With longer exposure, dysregulation of eNOS activity would result in apoptosis. The present study provides a molecular basis for the effects of eNOS in the prevention of endothelial cells apoptosis during early phase of high glucose exposure. These observations may contribute to the understanding of the pathogenesis of vascular complications in diabetes mellitus. © 2004 Wiley‐Liss, Inc.
Deregulation of DNA methyltransferases and loss of parental methylation at the insulin‐like growth factor II (Igf2)/H19 loci in p53 knockout mice prior to tumor development
Authors: Park In Young , Sohn Bo Hwa , Choo Jung Ha , Joe Cheol O. , Seong Je Kyung , Lee Young Ik , Chung Jae Hoon
Published: November 2004

To ascertain whether p53 deficiency in vivo leads to the deregulation of DNA methylation machinery prior to tumor development, we investigated the expression profile of DNA methyltransferases in the thymus and the liver of p53+/+, p53+/−, and p53−/− mice at 7 weeks of age before tumor development. The expression of DNA methyltransferases was examined in the thymus at 7 weeks of age, since the malignant T‐cell lymphoma develops most frequently in p53−/− mice around 20 weeks of age. Both mRNA and protein levels of Dnmt1 and Dnmt3b were increased in the thymus and the liver of p53‐deficient mice. The expression of Dnmt3a was also increased in the liver but not in the thymus of p53‐deficient mice. Dnmt3L expression was reduced in the thymus of p53+/− and p53−/− mice. The total 5‐methylcytosine (5‐MeC) in the genomic DNA of p53+/+, p53+/−, and p53−/− mice was quantitated by dot‐blot using antibody against 5‐MeC. Global methylation was increased in the thymus and the liver of p53‐deficient mice. To correlate the deregulated expression of DNA methyltransferases with the disturbance of the epigenetic integrity, we examined the DNA methylation of the imprinting control region (ICR) at the insulin‐like growth factor II (Igf2)/H19 loci in the thymus and the liver of p53+/+, p53+/−, and p53−/− mice. The region containing two CCCTC binding factor (CTCF) binding sites in the 5′‐ICR tended to be hypomethylated in the thymus of p53−/− mice, but not in the liver. The expression profile of Igf2 and H19 indicated that the thymus‐specific changes of Igf2 and H19 expression were coherent to the hypomethylation of the ICR in the thymus. Our results suggest that p53 is required for the maintenance of DNA methylation patterns in vivo. © 2004 Wiley‐Liss, Inc.
Co‐localization of poly(ADPR)polymerase 1 (PARP‐1) poly(ADPR)polymerase 2 (PARP‐2) and related proteins in rat testis nuclear matrix defined by chemical cross‐linking
Authors: Tramontano Filomena , Di Meglio Silvia , Quesada Piera
Published: November 2004

Poly(ADPR)polymerase 1 and 2 (PARP‐1, PARP‐2) are nuclear enzymes which function is based on specific interactions with DNA and nuclear proteins. PARPs targets include proteins involved in DNA replication, repair, and transcription and their function can be modulated either by protein–protein interaction with native PARP‐1 and by non‐covalent interaction with poly(ADP‐ribose) (pADPR) linked to the auto‐modified PARP‐1. Moreover, the association of pADPR and PARP‐1 with the nuclear matrix (NM) has been reported, based on the poly(ADP‐ribosyl)ation of nuclear matrix proteins (NMPs). In the present article, by the use of DNA and protein cross‐linking reactions, by cis‐diamminedichloroplatinum II (cDDP) and sodium tetrathionate (NaTT) respectively, we present more evidences about the association of PARP‐1, PARP‐2, and PARPs related proteins with the NM. Our findings confirmed that NM could be seen as a fraction greatly enriched in transcription factors (i.e., C/EBP‐β) and enzymes (DNA Topo II, DNA PK) that co‐localize with PARP‐1 and ‐2 at the matrix associated regions (MARs) of chromatin. Moreover, pADPR contributes to PARP‐1 localization at the NM, showing that PARP(s) activity co‐operates to the functions of this nuclear fraction. © 2004 Wiley‐Liss, Inc.
Osteoblasts lacking the vitamin D receptor display enhanced osteogenic potential in vitro
Authors: Sooy Karen , Sabbagh Yves , Demay Marie B.
Published: November 2004

1,25‐Dihydroxyvitamin D plays an important role in the regulation of osteoblast gene expression, regulating the expression of bone matrix proteins as well as that of Runx2, a key regulator of osteoblast differentiation. Studies in mice lacking the vitamin D receptor (VDR) have revealed that the actions of the VDR on the skeleton are not required in the setting of normal mineral ion homeostasis. Since paracrine and endocrine factors can compensate for gene defects in vivo, studies were performed to determine whether ablation of the VDR alters the program of osteoblast differentiation in vitro. Studies in primary calvarial cultures revealed that ablation of the VDR enhanced osteoblast differentiation. The cells from the VDR null mice exhibited an earlier onset and increased magnitude of alkaline phosphatase activity, as well as an earlier and sustained increase in mineralized matrix formation, demonstrating that this enhancement persists throughout the program of osteoblast differentiation. The expression of bone sialoprotein, which enhances mineralization, was also increased in the VDR null cultures. To determine whether the increase in osteoblast differentiation was associated with an increase in the number of osteogenic progenitors, the number of osteoblastic colony forming units (CFU‐OB) was evaluated. There was a twofold increase in the number of CFU‐OB in the cultures isolated from the VDR null mice. Furthermore, the VDR null CFU‐OB demonstrated an earlier onset and higher magnitude of expression of alkaline phosphatase activity when compared to the CFU‐OB from their wild‐type control littermates. These studies demonstrate that the VDR attenuates osteoblast differentiation in vitro and suggest that other endocrine and paracrine factors modulate the effect of the VDR on osteoblast differentiation in vivo. © 2004 Wiley‐Liss, Inc.
HHV‐6 and 7 DNA loads in lung tissues collected from patients with interstitial pneumonia
Authors: Yamamoto Keizo , Yoshikawa Tetsushi , Okamoto Souki , Yamaki Kenichi , Shimokata Kaoru , Nishiyama Yukihiro
Published: November 2004

The aim of this study is to determine whether human herpesvirus 6 (HHV‐6) and HHV‐7 might play an important role in causing interstitial pneumonia in patients who have not undergone transplantation. HHV‐6 and HHV‐7 DNAs were quantitated by real‐time polymerase chain reaction (PCR) in paraffin embedded lung tissues collected from 24 patients having the disease. Control tissues (without fibrosis) were also collected from 19 of the 24 patients. Statistical analysis was carried out by the Wilcoxon signed rank test or the Mann–Whitney U‐test. HHV‐6 DNA was detected in 3 (12.5%) of the 24 target tissues and 3 (15.8%) of the 19 control tissues, respectively. In contrast, HHV‐7 DNA was detected in 19 (79.2%) of the 24 target tissues and 11 (57.9%) of the 19 control tissues. Neither HHV‐6 DNA load (P = 0.6395) nor HHV‐7 DNA load (P = 0.5966) in target tissues differed between males and females. Neither HHV‐6 DNA load (P = 0.9589) nor HHV‐7 DNA load (P = 0.7419) in target tissues differed between cases with and without underlying collagen disease. While HHV‐6 DNA load did not differ between the target and control tissues (P > 0.9999), the HHV‐7 DNA load was significantly higher in the target tissue than in the control tissue (P = 0.0298). This study suggests that HHV‐7 may play an important role in causing interstitial pneumonia in patients who are not transplant recipients. J. Med. Virol. 75:70–75, 2005. © 2005 Wiley‐Liss, Inc.
Laboratory diagnosis and surveillance of human respiratory viruses by PCR in Victoria, Australia, 2002–2003
Authors: Druce Julian , Tran Thomas , Kelly Heath , Kaye Matthew , Chibo Doris , Kostecki Renata , Amiri Abdul , Catton Mike , Birch Chris
Published: November 2004

Respiratory viruses were identified by the polymerase chain reaction (PCR) in more than 4,200 specimens collected during 2002 and 2003 in Victoria, Australia from patients admitted to hospitals or participating in an influenza surveillance program. Influenza viruses and picornaviruses were important causes of morbidity in both years. Additional testing of picornavirus‐positive samples suggested that rhinoviruses but not enteroviruses were more likely to be associated with respiratory symptoms, irrespective of the season in which they circulated. Detection of influenza viruses was strongly associated with the clinical symptoms of cough, fever, and fatigue; but each of the other respiratory viruses occasionally caused these symptoms or was responsible for symptoms severe enough to require hospitalization. Human coronaviruses HCoV‐OC43 and HCoV‐229E circulated at low levels throughout the study period with peak activity in winter, but overall did not circulate as widely as has often been reported for these agents. Evidence for the human metapneumovirus (hMPV) was only sought in the second year of the study and revealed low‐level circulation of this virus, mainly in the cooler months among the very young and adult populations. The detection rate of all viruses declined with increasing age of the patient, particularly in hospital patients. Infection with more than one respiratory virus occurred in a small number of patients; picornaviruses were most commonly implicated in these dual infections. J. Med. Virol. 75:122–129, 2005. © 2005 Wiley‐Liss, Inc.
Measles virus genotype circulation in Spain after implementation of the national measles elimination plan 2001–2003
Authors: Mosquera María Mar , Ory Fernando de , Echevarría Juan Emilio ,   The Network of Laboratories in the Spanish National Measles Elimination Plan
Published: November 2004

Molecular characterization of measles virus is important for disease surveillance and for monitoring elimination of the virus throughout the world. Furthermore, knowledge of genotype distribution in as many countries as possible, is useful for tracing the origin of a strain, especially in countries without endemic measles disease, where most cases are imported. Data on genotypes circulating in Spain from 1970 to 1997 showed the prevalence of genotypes C1, C2, and D6, with subsequent replacement of each other. After the establishment of the Spanish Measles Elimination Plan, genotyping with a new retrotranscriptase polymerase chain reaction (RT‐PCR) was undertaken directly on 92 specimens, corresponding to 90 patients, which were positive for measles by a different diagnostic RT‐PCR. Genotypes B3, D4, D8, A, C2, H1, and D7 were found in different autonomous communities (Madrid, Balearic Islands, Valencia Community, Extremadura, Andalusia, Canary Islands and Murcia) between 2001 and 2003 with none of these genotypes being prevalent. After the introduction of the vaccine in 1978, the incidence of the disease decreased from 150,000 cases in 1977 to 64 in 2002. This could be the reason for the change observed in the pattern of measles genotype circulation, since this pattern was reported in countries at an advanced stage of eradication of measles. This report considers that Spain is on the way to eradicating measles. J. Med. Virol. 75:137–146, 2005. © 2005 Wiley‐Liss, Inc.
ADAM23 is a cell‐surface glycoprotein expressed by central nervous system neurons
Authors: Goldsmith Alexander P. , Gossage Samuel J. , ffrench‐Constant Charles
Published: November 2004

Several members of the ADAM (a disintegrin and metalloprotease) family of proteins have been implicated in biological processes ranging from fertilization to myoblast fusion and neural cell fate determination. These proteins have so far been studied mostly in terms of their protease activity, but a considerable amount of evidence suggests that many ADAMs are also important as receptors for cell‐surface integrins. We have shown that, for one such member of the family, ADAM23, mRNA transcripts are expressed in neuronal cells throughout the rat brain, at all stages of postnatal development, and that particularly high transcript concentrations are found in the hippocampus and cerebellum. Using an antibody that we raised against the rat ADAM23 disintegrin domain, we found that ADAM23 is present at detectable levels only in nervous system tissue. Our analysis of ADAM23 expression in cultured cerebellar granule cells (CGCs) furthermore suggested that this protein is synthesized as a glycosylated precursor of about 100 kD whose maturation depends on cleavage by furin or a related enzyme. We have also shown ADAM23 to be expressed primarily as a cell‐surface protein that appears to be localized to sites of intercellular contact. Taken together, these data are consistent with a model wherein ADAM23 serves to mediate cell–cell interactions within the mammalian CNS. © 2004 Wiley‐Liss, Inc.
Increased GABAergic function in mouse models of Huntington's disease: Reversal by BDNF
Authors: Cepeda Carlos , Starling Amaal J. , Wu Nanping , Nguyen Oanh K. , Uzgil Besim , Soda Takahiro , André Veronique M. , Ariano Marjorie A. , Levine Michael S.
Published: November 2004

Huntington's disease (HD) is characterized by loss of striatal γ‐aminobutyric acid (GABA)ergic medium‐sized spiny projection neurons (MSSNs), whereas some classes of striatal interneurons are relatively spared. Striatal interneurons provide most of the inhibitory synaptic input to MSSNs and use GABA as their neurotransmitter. We reported previously alterations in glutamatergic synaptic activity in the R6/2 and R6/1 mouse models of HD. In the present study, we used whole‐cell voltage clamp recordings to examine GABAergic synaptic currents in MSSNs from striatal slices in these two mouse models compared to those in age‐matched control littermates. The frequency of spontaneous GABAergic synaptic currents was increased significantly in MSSNs from R6/2 transgenics starting around 5–7 weeks (when the overt behavioral phenotype begins) and continuing in 9–14‐week‐old mice. A similar increase was observed in 12–15‐month‐old R6/1 transgenics. Bath application of brain‐derived neurotrophic factor, which is downregulated in HD, significantly reduced the frequency of spontaneous GABAergic synaptic currents in MSSNs from R6/2 but not control mice at 9–14 weeks. Increased GABA current densities also occurred in acutely isolated MSSNs from R6/2 animals. Immunofluorescence demonstrated increased expression of the ubiquitous α1 subunit of GABAA receptors in MSSNs from R6/2 animals. These results indicate that increases in spontaneous GABAergic synaptic currents and postsynaptic receptor function occur in parallel to progressive decreases in glutamatergic inputs to MSSNs. In conjunction, both changes will severely alter striatal outputs to target areas involved in the control of movement. © 2004 Wiley‐Liss, Inc.
Neurotoxic prostaglandin J2 enhances cyclooxygenase‐2 expression in neuronal cells through the p38MAPK pathway: A death wish?
Authors: Li Zongmin , Jansen Marlon , Ogburn Kenyon , Salvatierra Laura , Hunter Lavonne , Mathew Sneha , Figueiredo‐Pereira Maria E.
Published: November 2004

The role of the proinflammatory and inducible form of cyclooxygenases (COX‐2) in neurodegeneration is not well defined. Some of its metabolic products, such as prostaglandins (PG) of the J2 series, are known to be neurotoxic. Here we demonstrate that PGJ2 enhances COX‐2 gene expression without elevating COX‐1 levels in neuronal cells. PGJ2 also increased PGE2 production, establishing that the de novo synthesized COX‐2 is enzymatically active. PGJ2 derivatives, such as 15d‐PGJ2, are known activators of PPARγ, a nuclear receptor that activates gene expression. However, the selective PPARγ agonist ciglitazone failed to up‐regulate COX‐2, indicating that the PGJ2 effect on COX‐2 is PPARγ independent. Furthermore, PGJ2 stabilized IκBα levels, indicating that NFκB is not active under these conditions. The blocking of neuronal NFκB activity by PGJ2 may be an important contributor to its neurotoxicity, insofar as NFκB transactivation seems to be required for neuronal survival in the CNS. Interleukin‐1 (IL1) is a proinflammatory cytokine known to stimulate the expression of genes associated with inflammation, including COX‐2. Notably, IL1 mRNA levels in the neuronal cells were increased by PGJ2 treatment. The proinflammatory cytokine may mediate COX‐2 up‐regulation by PGJ2 through p38MAPK and not JNK activation, in that only an inhibitor of the former prevented the COX‐2 increase. Thiol‐reducing agents, such as N‐acetylcysteine, protected the neuronal cells from the deleterious effects of PGJ2, whereas ascorbic acid did not. Collectively, our findings suggest that proinflammatory conditions that lead to COX‐2 up‐regulation and the concomitant production of PGJ2 initiate a mechanism of self‐destruction through an autotoxic loop between PGJ2 and COX‐2 that may exacerbate neurodegeneration beyond a point of no return. Thiol‐reducing antioxidants may offer an optimal strategy for halting this neurodegenerative process. © 2004 Wiley‐Liss, Inc.
Middle East climate simulation and dominant precipitation processes
Authors: Evans Jason P. , Smith Ronald B. , Oglesby Robert J.
Published: November 2004

The ability of both the ECMWF‐TOGA analyses and regional model RegCM2 to simulate the climate of the Middle East is examined. The climate of the region displays high spatial, seasonal and interannual variability, providing a strong test of a climate model's abilities. The higher resolution of RegCM2, compared with the ECMWF analyses, allows it to capture the spatial variability of temperature and precipitation better despite model biases being present. Both RegCM2 and the analyses have a cold bias, exacerbated in RegCM2 by a bias present in the prescribed sea‐surface‐temperature forcing. RegCM2 does not capture the annual cycle of precipitation on the Black and Caspian Sea coasts, where very steep topography exists, nor on the eastern Mediterranean coast, where the coastal mountains are not resolved. RegCM2 does capture the seasonal cycle in the Fertile Crescent and Zagros Mountains, where it is strongly influenced by a plateau circulation above the Iranian plateau. It is shown that accurate simulation of precipitation in these regions requires the correct simulation of storm tracks, topographic interactions and atmospheric stability. RegCM2 is better able to simulate the interannual variability averaged over the entire domain compared with the ECMWF analyses; however, they both have difficulty reproducing the interannual variability in particular subregions. It is also shown that processes controlling the seasonality of precipitation differ in different subregions and are often different from the processes controlling interannual variability. This suggests that, in order to model precipitation successfully, a horizontal scale that allows differentiation of precipitation zones dominated by different precipitation processes, both seasonally and interannually, is required. Copyright © 2004 Royal Meteorological Society
Study of the adsorption of benzimidazole and 2‐mercaptobenzothiazole on an iron surface by confocal micro‐Raman spectroscopy
Authors: Wang G. , Harrison A. , Li X. , Whittaker G. , Shi J. , Wang X. , Yang H. , Cao P. , Zhang Z.
Published: November 2004

Benzimidazole (BIMH) and 2‐mercaptobenzothiazole (HMBT) dissolved in ethanol were chosen for the investigation of the interaction between organic molecules and surface atoms of iron or iron oxide by confocal micro‐Raman spectroscopy. Both BIMH and HMBT show enhanced and highly structured spectra in the 200–1500 cm−1 region when the iron is at a potential of ca −0.7 to −0.9 V in a neutral medium. BIMH had a weak interaction with the iron surface in a basic medium but it was chemically adsorbed in a neutral medium. HMBT was chemically adsorbed on the iron via the exocyclic S and N atoms in acidic and neutral solutions, whereas in basic media it was bound electrostatically. Copyright © 2004 John Wiley & Sons, Ltd.
Theory of laser radiation scattering in integrated optical
waveguide with 3D-irregularities in presence of noise: vector
consideration
Authors: Egorov A A
Published: November 2004

Vector theory of the waveguide laser irradiation
scattering in an integrated optical waveguide with
3D-irregularities in the presence of a noise is described. For
simplification of the analysis of the problem we neglected
cross-correlation connections, both between irregularities of the
boundaries of the waveguide, and between irregularities of the
boundaries and heterogeneities of the refraction index of
waveguide mediums. The approximate solution of inhomogeneous
three-dimensional wave equation is derived by the Green function
method. The analytical formulas of the radiating fields for the
propagating and evanescent modes are fined. Physico-mathematical
interpretation of derived expressions is given.
New Developments in Photoinitiators
Authors: Dietliker Kurt , Jung Tunja , Benkhoff Johannes , Kura Hisatoshi , Matsumoto Akira , Oka Hidetaka , Hristova Daniela , Gescheidt Georg , Rist Günther
Published: November 2004

New developments of photoinitiators are the base for the further extension of the radiation curing technology. Three different approaches are presented: The optimized combination of a bisacylphosphine oxide photoinitiator with a suitable stabilizer package allows the UV curing of clear coatings with an excellent performance in outdoor use. The design of new oxime ester derivatives allowed the introduction of a new tailor‐made photoinitiator for color filter resists applications that considerably improves the color quality of the color filter. A novel photoinitiator class capable of releasing strong amine bases opens new opportunities for expanding radiation curing into resins types that crosslink by base‐catalyzed processes.
Water Soluble, Photocurable Resins for Rapid Prototyping Applications
Authors: Stampfl J. , Wöß A. , Seidler S. , Fouad H. , Pisaipan A. , Schwager F. , Liska R.
Published: November 2004

Rapid Prototyping (RP) is a suitable manufacturing method for fabricating structures with high geometric complexity and heavily undercut features. A special class of such structures which cannot easily be fabricated with traditional manufacturing methods are cellular materials [1].

Rapid Prototyping allows the fabrication of cellular materials on a similar size scale like in natural material‐structures (e.g. trabecular bone). By using appropriate moulding techniques, these structures can be fabricated out of a wide variety of materials (polymers, ceramics, composites). In this work, several RP techniques are investigated regarding their suitability for the fabrication of cellular solids. The main focus is on using digital light projection (DLP, a variant of stereolithography) in combination with gelcasting [2] as a moulding technique. Besides using commercial light‐sensitive resins, a class of newly developed water soluble resins has been evaluated regarding its usability as sacrificial mould material. These water soluble resins are compatible with a wide range of moulding techniques and therefore offer new routes for the fabrication of complex shapes out of more advanced materials than it is possible with currently used manufacturing techniques.
Bifunctional Adhesion Promoter for Grafting Polypyrrole Films on Metal/Metal Oxide Surfaces
Authors: Oberoi Sonia , Jähne Evelin , Adler Hans‐Jürgen P.
Published: November 2004

A new class of pyrrole derivatives, ω‐[(3‐phenyl) pyrrol‐1‐yl] alkyl phosphonic acids with long chains of 10 and 12 carbon atoms were synthesised to graft polypyrrole layers on metal/metal oxide surfaces. These compounds are bifunctional containing two reactive moieties, pyrrole as the polymerisable group and phosphonic acid as the anchoring group. Contact angle measurements and X‐ray photoelectron spectroscopy (XPS) confirmed adsorption with phosphonic acid group attached to the surface. Surface plasmon resonance (SPR) spectroscopy indicated that adsorption starts in seconds and is completed in few hours. Adsorption is followed by surface induced polymerisation with further monomer. We obtained dense and homogeneous polypyrrole films, which were characterised for their morphology and thickness by atomic force microscopy (AFM). The derivatives form a strongly bonded composite of metal with polymer.