bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative anatomy of the foramen ovale in the hearts of cetaceans
Authors: Macdonald Alastair A. , Carr Peter A. , Currie Richard J. W.
Published: May 2007

The structure of the cardiac foramen ovale from 17 species representing six cetacean families, the Monodontidae, Phocoenidae, Delphinidae, Ziphiidae, Balaenidae and the Balaenopteridae, was studied using the scanning electron microscope. Eight white whale fetuses (Delphinapterus leucas) and a narwhal fetus (Monodon monoceros) represented the Monodontidae; one fetal and nine neonatal harbour porpoises (Phocoena phocoena) and a finless porpoise fetus (Neophocoena phocoenoides) represented the Phocoenidae; two white‐beaked dolphin fetuses (Lagenorhynchus albirostris), four fetal and one neonatal Atlantic white‐sided dolphins (Lagenorhynchus acutus), a Risso's dolphin fetus (Grampus griseus), two common bottle‐nosed dolphin neonates (Tursiops truncatus), a female short‐beaked common dolphin fetus (Delphinus delphis), four killer whale fetuses (Orcinus orca) and two long‐finned pilot whale fetuses (Globicephala melas) represented the Delphinidae; two northern bottlenose whale fetuses (Hyperoodon ampullatus) represented the Ziphiidae; one bowhead whale fetus (Balaena mysticetus) represented the Balaenidae and five Common minke whale fetuses (Balaenoptera acutorostrata), one blue whale fetus (Balaenoptera musculus), nine fin whale fetuses (Balaenoptera physalus) and four humpback whale fetuses (Megaptera novaeangliae) represented the Balaenopteridae. The hearts of an additional two incompletely identified toothed and four baleen whale fetuses were also studied. In each species the fold of tissue derived from the cardiac septum primum and subtended by the foramen ovale had the appearance of a short tunnel or sleeve which was fenestrated at its distal end. In the toothed whales the tissue fold was tunnel‐shaped with the interatrial septum as the floor whereas in baleen whales it was more sleeve‐like. In toothed whales thin threads extended from the fold to insert into the interatrial septum whereas a network of threads covered the distal end of the sleeve in the baleen whales. Similar structures were present in the corresponding cardiac tissues of neonatal Hippopotamidae.
Epidermal differentiation in embryos of the tuatara Sphenodon punctatus (Reptilia, Sphenodontidae) in comparison with the epidermis of other reptiles
Authors: Alibardi L. , Gill B. J.
Published: May 2007

Studying the epidermis in primitive reptiles can provide clues regarding evolution of the epidermis during land adaptation in vertebrates. With this aim, the development of the skin of the relatively primitive reptile Sphenodon punctatus in representative embryonic stages was studied by light and electron microscopy and compared with that of other reptiles previously studied. The dermis organizes into a superficial and deep portion when the epidermis starts to form the first layers. At embryonic stages comparable with those of lizards, only one layer of the inner periderm is formed beneath the outer periderm. This also occurs in lizards and snakes so far studied. The outer and inner periderm form the embryonic epidermis and accumulate thick, coarse filaments (25–30 nm thick) and sparse alpha‐keratin filaments as in other reptiles. Beneath the embryonic epidermis an oberhautchen and beta‐cells form small horny tips that represent overlapping borders along the margin of beta‐cells that overlap other beta‐cells (in a tile‐like arrangement). The tips resemble those of agamine lizards but at a small scale, forming a lamellate‐spinulated pattern as previously described in adult epidermis. The embryonic epidermis matures by the dispersion of coarse filaments among keratin at the end of embryonic development and is shed around hatching. The presence of these matrix organelles in the embryonic epidermis of this primitive reptile further indicates that amniote epidermis acquired interkeratin matrix proteins early for land adaptation. Unlike the condition in lizards and snakes, a shedding complex is not formed in the epidermis of embryonic S. punctatus that is like that of the adult. Therefore, as in chelonians and crocodilians, the epidermis of S. punctatus also represents an initial stage that preceded the evolution of the shedding complex for moulting.
µMRI–HREM pipeline for high‐throughput, high‐resolution phenotyping of murine embryos
Authors: Pieles Guido , Geyer Stefan H. , Szumska Dorota , Schneider Jürgen , Neubauer Stefan , Clarke Kieran , Dorfmeister Karl , Franklyn Angela , Brown Steve D. , Bhattacharya Shoumo , Weninger Wolfgang J.
Published: May 2007

Rapid and precise phenotyping analysis of large numbers of wild‐type and mutant mouse embryos is essential for characterizing the genetic and epigenetic factors regulating embryogenesis. We present a novel methodology that permits precise high‐throughput screening of the phenotype of embryos with both targeted and randomly generated mutations. To demonstrate the potential of this methodology we show embryo phenotyping results produced in a large‐scale ENU‐mutagenesis study. In essence this represents an analysis pipeline, which starts with simultaneous micro‐magentic resonance imaging (µMRI) screening (voxel size: 25.4 × 25.4 × 24.4 µm) of 32 embryos in one run. Embryos with an indistinct phenotype are then cut into parts and suspect organs and structures are analysed with HREM (high‐resolution episcopic microscopy). HREM is an imaging technique that employs ‘positive’ eosin staining and episcopic imaging for generating three‐dimensional (3D) high‐resolution (voxel size: 1.07 × 1.07 × 2 µm) digital data of near histological contrast and quality. The results show that our method guarantees the rapid availability of comprehensive phenotype information for high numbers of embryos in, if necessary, histological quality and detail. The combination of high‐throughput µMRI with HREM provides an alternative screening pipeline with advantages over existing 3D phenotype screening methods as well as traditional histology. Thus, the µMRI‐HREM phenotype analysis pipeline recommends itself as a routine tool for analysing the phenotype of transgenic and mutant embryos.
Rituximab for warm‐type idiopathic autoimmune hemolytic anemia: a retrospective study of 11 adult patients
Authors: D’Arena Giovanni , Califano Catello , Annunziata Mario , Tartarone Alfredo , Capalbo Silvana , Villani Oreste , Amendola Giovanni , Pietrantuono Giuseppe , Ferrara Felicetto , Pinto Antonio , Musto Pellegrino , D’Arco Alfonso Maria , Cascavilla Nicola
Published: May 2007

Warm‐type idiopathic autoimmune hemolytic anemia (AIHA) is a relatively common hematologic disorder resulting from autoantibody production against red blood cells. Steroids represent the first‐line therapeutic option, and immunosuppressive agents as well as splenectomy are used for refractory cases. Recently, the anti‐CD20 monoclonal antibody rituximab has been shown to control autoimmune hemolysis in patients with refractory chronic disease. We report results from a retrospective analysis of 11 adult patients receiving rituximab for steroid‐refractory AIHA of the warm type at a mean age of 55 yr (range 23–81 yr). All patients were given methyl‐prednisolone as first‐line treatment and some of them also received azathioprine and intravenous high‐dose immunoglobulins. One patient underwent splenectomy. All patients were considered refractory to steroids and/or immunosuppressive drugs and all were then given weekly rituximab (375 mg/m2) for four consecutive weeks. An increase in hemoglobin (Hgb) levels in response to rituximab, with a mean increment of 3.3 g/dL (95% CI 2.1–4.4), was observed in all cases. Four patients required packed red cell transfusions before starting rituximab and all became transfusion‐free. At a mean follow‐up of 604 d (range 30–2884 d) since the treatment of AIHA with rituximab, all patients are alive, eight (73%) of them in complete remission (CR) and three (27%) in partial remission (PR). A moderate hemolysis still persisted in six (54%) patients. In conclusion, our experience clearly demonstrates that anti‐CD20 monoclonal antibody rituximab is an effective and safe alternative treatment option for idiopathic AIHA, in particular, for steroid‐refractory disease.
The effect of ranitidine on antibody responses to polysaccharide vaccines in patients with B‐cell chronic lymphocytic leukaemia
Authors: Van der Velden Ankie M. T. , Van Velzen‐Blad Heleen , Claessen Anke M. E. , Van der Griend René , Oltmans Ruud , Rijkers Ger T. , Biesma Douwe H.
Published: May 2007

Objectives: To analyse the effects of ranitidine treatment on vaccination induced antibody responses in patients with chronic lymphocytic leukaemia (CLL).Methods: Fifty CLL patients were vaccinated with tetanus conjugated Hib vaccine and a 23‐valent pneumococcal polysaccharide vaccine with (n = 25) or without (n = 25) ranitidine treatment in a matched case‐‐control setting. Anti tetanus toxoid (TT), anti‐Hib and anti‐pneumococcal antibody levels were determined before and after vaccination. Additionally, cytokine levels were assessed in patients treated with ranitidine.Results: Vaccination‐induced increases in anti‐Hib and anti‐TT antibody levels were higher in the ranitidine group compared with the control group. Anti‐pneumococcal antibody responses were not improved by administration of ranitidine. Higher levels of IL‐18 were found in patients treated with ranitidine compared with healthy controls. Levels of IL‐6, IL‐8, IL‐18, RANTES, IP‐10, sVCAM‐1 and sICAM‐1 were within normal ranges and did not change during ranitidine treatment.Conclusion: Ranitidine treatment improves vaccination‐induced T‐cell dependent antibody responses in patients with CLL but has no beneficial effect on the response to vaccination with unconjugated polysaccharide antigens.
Immunophenotypic analysis of Epstein–Barr virus (EBV)‐infected CD8+ T cells in a patient with EBV‐associated hemophagocytic lymphohistiocytosis
Authors: Wada Taizo , Kurokawa Toshiro , Toma Tomoko , Shibata Fumie , Tone Yumi , Hashida Yoko , Kaya Hiroyasu , Yoshida Takashi , Yachie Akihiro
Published: May 2007

Hemophagocytic lymphohistiocytosis (HLH) is a severe and often fatal condition characterized by uncontrolled activation of T cells and macrophages. In Epstein–Barr virus (EBV)‐associated HLH (EBV‐HLH), the pathogenic roles of ectopic EBV infection in the T‐cell population and of clonal proliferation of EBV‐infected T cells has been described. However, the immunophenotype of EBV‐infected T cells has not been fully characterized. Here we describe a case of EBV‐HLH presenting with a massive clonal proliferation of CD8+ T cells with TCR VB14. Analysis of in situ hybridization for EBV‐encoded small RNA1 showed that only CD8+ T cells harbored EBV in this patient. The EBV‐infected TCR VB14+ CD8+ T cells exhibited unique immunophenotypic features including lacked CD5 expression and a markedly bright expression of HLA‐DR. After initiation of treatment with prednisolone, etoposide, and cyclosporin A, the percentage of infected cells declined progressively in parallel with other serum markers such as ferritin. These findings suggest that lacking expression of CD5 on CD8+ T cells with specific TCR VB may serve as a useful marker of dysregulated T‐cell activation and proliferation in EBV‐HLH.
QUASI analysis of the HIV-1 envelope sequences in the Los Alamos National Laboratory HIV sequence database: pattern and distribution of positive selection sites and their frequencies over yearsThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Liang Binhua , Luo Ma , Ball T. Blake , Plummer Francis A.
Published: May 2007

The envelope (env) protein of human immunodeficiency virus type 1 (HIV-1) plays a crucial role in virus entry and is a central target for HIV vaccine design. Using the QUASI program, we analyzed the conserved regions of all currently available env sequences in the Los Alamos National Laboratory HIV Sequence Database and identified positive selection (PS) sites that are likely to be restricted by host immune responses. We found that PS sites are dispersed across conserved regions of env sequence, and that the C3, C4, and C5 regions were the most targeted. Several regions were identified as being PS free and were mainly distributed in the C1 and C2 regions. When comparing individual QUASI PS site frequencies across clades and geographical regions with the overall frequency of the entire env database, the env sequences from North America showed significantly lower PS site frequency, while those from Asia were significantly higher using Student's t test. The QUASI PS site frequency of env proteins from viruses isolated from different years showed that the PS site frequencies of the env population increased over time. Our study provides an overview of PS sites across the conserved regions of HIV-1 env sequences.
Effects of estradiol and 4-hydroxytamoxifen on the conformation, thermal stability, and DNA recognition of estrogen receptor β
Authors: Vijayanathan Veena , Greenfield Norma J. , Thomas T. J. , Ivanova Margarita M. , Tyulmenkov Valentyn V. , Klinge Carolyn M. , Gallo Michael A. , Thomas Thresia
Published: May 2007

Estrogen receptors (ERα and ERβ) are ligand-activated transcription factors. We examined the effects of estradiol (E2), 4-hydroxytamoxifen (HT), and the estrogen response element (ERE) on the helical content and thermal unfolding of ERβ. A circular dichroism (CD) spectrum of ERβ showed changes at 210 and 222 nm that were due to the presence of E2, which is indicative of partial unfolding. In contrast, HT did not alter the CD spectrum of ERβ. The addition of E2 + ERE caused an increase in the α-helical content and an increase in the temperature midpoint of folding transition (TM) from 39 ± 0.7 °C to 57.2 ± 1 °C. The addition of E2 + mutant ERE, or E2 + control oligonucleotide, increased the TMof ERβ to 45 ± 2 °C only. In the presence of HT, ERβ yielded similar TMvalues (55–58 °C) with ERE, mutant ERE, or control oligodeoxynucleotide. The binding affinity of ERβ for ERE increased 125.7-fold as a result of the presence of E2, but only 4-fold as a result of HT. These results demonstrate coupled effects of E2and ERE on ERβ stability and binding affinity. The increased thermal stability of HT–ERβ–ERE was associated with reduced specificity of ERβ–ERE recognition, illustrating profound differences in conformational states of ERβ induced by E2and HT.
BMPs regulate differentiation of a putative visceral endoderm layer within human embryonic stem-cell-derived embryoid bodies
Authors: Conley Brock J , Ellis Sarah , Gulluyan Lerna , Mollard Richard
Published: May 2007

Human embryonic stem cells (HESCs), pluripotent cells derived from the inner cell mass (ICM) of human blastocysts, represent a novel tool for the study of early human developmental events. When cultured in suspension with serum, HESCs form spherical structures resembling embryoid bodies (EBs). We show that differentiation of HESCs within EBs occurs radially, with central cells then undergoing apoptosis in association with EB cavitation. Cells within the outer layer of cavitating EBs display stage-specific immunoreactivity to pan-keratin, cytokeratin-8, GATA6, α-fetoprotein, and transthyretin specific antibodies, and hybridization to disabled-2, GATA4, and GATA6 specific riboprobes. Transmission electron microscopy of these cells reveals clathrin-coated micropinocytotic vesicles, microvilli, and many vacuoles, a phenotype consistent with mouse visceral endoderm (VE) rather than mouse definitive or parietal endoderm. When cultured in media supplemented with the BMP inhibitor noggin, or in the absence of serum, HESC derivatives do not develop the mouse VE-like phenotype. The addition of BMP-4 to noggin-treated HESCs cultured in serum or in serum-free conditions reconstituted development of the VE-like phenotype. These data demonstrate that human EBs undergo developmental events similar to those of mouse EBs and that in vitro BMP signalling induces derivatives of the human ICM to express a phenotype similar to mouse VE.
The inhibitory effect of simvastatin on the ADMA-induced inflammatory reaction is mediated by MAPK pathways in endothelial cells
Authors: Jiang Jun-Lin , Wang Shan , Li Nian-Sheng , Zhang Xiao-Hong , Deng Han-Wu , Li Yuan-Jian
Published: May 2007

Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, is emerging as a key contributor for endothelial dysfunction associated with inflammation. Statins can inhibit vascular inflammatory reaction and improve endothelial function. The aim of this study was to investigate in human endothelial cells the signaling pathways of ADMA-induced inflammatory reaction and potential inhibitory effects of simvastatin. Endothelial cells were cultured and used for all of the studies. Tumor necrosis factor-α (TNF-α) and soluble intercellular adhesion molecule-1 (sICAM-1) were determined by enzyme-linked immunosorbent assay. Nuclear factor-κB (NF-κB) was assayed by electrophoretic mobility shift assay. The activation of mitogen-activated protein kinases (MAPKs), including p38 MAPK and extracellular signal-related kinase (ERK1/2), were characterized by Western blot analysis. Treatment with ADMA (3–30 µmol/L) increased the concentration of sICAM-1 in a dose-dependent manner. ADMA (30 µmol/L) significantly enhanced the concentrations of TNF-α and sICAM-1, the activity of NF-κB and the phosphorylation of p38 MAPK and ERK1/2. The increased secretion of TNF-α and sICAM-1 and the increased activity of NF-κB by ADMA were altered by SB203580 (5 µmol/L) or PD98059 (20 µmol/L), but not by LY294002 (20 µmol/L). Simvastatin (0.1, 0.5, or 2.5 µmol/L) markedly inhibited the elevated concentrations of TNF-α and sICAM-1, the activity of NF-κB, and the phosphorylation of p38 MAPK and ERK1/2 induced by ADMA. Simvastatin inhibited ADMA-induced inflammatory reaction by p38 MAPK and ERK1/2 pathways in cultured endothelial cells.
Mitochondria-independent morphological and biochemical apoptotic alterations promoted by the anti-tumor agent bleomycin inSaccharomyces cerevisiae
Authors: Aouida Mustapha , Mekid Halima , Belhadj Omrane , Mir Lluis M. , Tounekti Omar
Published: May 2007

Bleomycin is a highly potent cytotoxic and genotoxic agent used in the chemotherapy of various types of tumors. It is a radiomimetic anticancer drug that produces single- and double-stranded DNA breaks in a catalytic way. Using Saccharomyces cerevisiae as a model system, we show that when a high amount of bleomycin molecules is internalized (100 µmol/L), morphological changes identical to those usually associated with apoptosis, i.e., a sub-G1region peak, chromatin condensation, and very rapid DNA fragmentation into oligonucleosomal-sized fragments, are observed. The known bleomycin inhibitors cobalt and EDTA were able to prevent bleomycin nucleasic activity and thus apoptotic cell death. However, both oligomycin, a potent inhibitor of the mitochondial F0F1-ATPase, and antimycin, a drug affecting mitochondria respiration, were unable to prevent the bleomycin-induced apoptotic-like cell death. These results suggest that high bleomycin concentrations induce an apoptosis-like mitochondria-independent cell death in yeast.
Novel irreversible caspase-1 inhibitor attenuates the maturation of intracellular interleukin-1β
Authors: Ma Xue-Qin , Zhang Hua-Jie , Zhang Ya-Hui , Chen Yi-Hua , Wu Fang , Du Jun-Qing , Yu Hai-Ping , Zhou Zhong-Liang , Li Jing-Ya , Nan Fa-Jun , Li Jia
Published: May 2007

. Caspase-1, the most efficient enzyme in processing the proinflammatory cytokines interleukin 1β and interleukin 18 in humans, is associated with inflammatory diseases such as rheumatoid arthritis, osteoarthritis, and some neuronal diseases. We previously reported that isoquinoline-1,3,4-trione and its derivatives are novel caspase-3 inhibitors that could attenuate apoptosis in vitro and in vivo. Here we report a novel derivative of isoquinoline-1,3,4-trione that is highly potent in inhibiting caspase-1 activity in an irreversible and slow-binding manner, thus inhibiting cellular caspase-1 activity and the maturation of interleukin 1β in U-937 cells.
A novelAmoeba proteus120 kDa actin-binding protein with only 1 filamin repeat and a coiled-coil region
Authors: Sobczak Magdalena , Kocik Elżbieta , Rędowicz Maria Jolanta
Published: May 2007

A novel 120 kDa actin-binding protein (ApABP-F1) was found in Amoeba proteus. It was distributed throughout the cytoplasm, mainly in the subplasma membrane and perinuclear–nuclear areas, enriched in actin. The full-length cDNA of ApABP consisted of 2672 nucleotides with an open reading frame of 878 amino acids, giving a ∼95 kDa protein with a theoretical pI value of 5.11. It had a novel domain organization pattern: the N terminus (residues 1–104) contained 1 calponin-homology (CH) domain, followed by only 1 region that was homologous to the filamin repeat (FR, residues 209–324), and a central region (residues 344–577) exhibiting a very high probability of coiled-coil formation, probably engaged in the observed protein dimerization. A phylogenetic tree constructed for CH domains from 25 various proteins revealed that the CH domain of ApABP was most related to that of the hypothetical mouse KIAA0903-like protein, whereas not much relationship to either filamins or the gelation factor (ABP-120) of Dictyostelium discoideum and Entamoeba histolytica was found.
A 27.368 kDa retinal reductase in New Zealand white rabbit liver cytosol encoded by the peroxisomal retinol dehydrogenase–reductase cDNA: purification and characterization of the enzymeThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Du Kun , Liu Ge-Fei , Xie Jian-Ping , Song Xu-Hong , Li Rui , Liang Bin , Huang Dong-Yang
Published: May 2007

We obtained a full-length cDNA based on a sequence deposited in GenBank (accession No. AB045133), annotated as rabbit peroxisomal NADP(H)-dependent retinol dehydrogenase–reductase (NDRD). The rabbit NDRD gene, like its mouse and human homologs, harbors 2 initiation sites, one of which theoretically encodes a 29.6 kDa protein with 279 amino acids, and the other encodes a 27.4 kDa protein with 260 amino acids. The purification of a rabbit cytosolic retinol oxidoreductase with a subunit molecular mass of 34 kDa and an N terminus that is not completely identical to that of NDRD, has been reported. An enzyme responsible for the all-trans retinal reductase activity in the liver cytosol of New Zealand white rabbit was purified to homogeneity using differential centrifugation and successive chromatographic analyses. The subunit molecular mass of the purified enzyme, revealed by SDS–PAGE, was approximately 27 kDa. The intact molecular mass, measured by MALDI–TOF mass spectrometry, was 27.368 kDa. The 60 kDa relative mobility observed in size-exclusion chromatography indicates that the native protein probably exists as a dimer. The purified enzyme was positively confirmed to be the product of NDRD by peptide mass fingerprinting, tandem mass spectrometry, and N-terminal sequencing. Taken together, the results suggested that the native protein is truncated at the N terminus.
Differential effects of c-Ras upon transformation, adipocytic differentiation, and apoptosis mediated by the simian virus 40 large tumor antigen
Authors: Cao Jun , Arulanandam Rozanne , Vultur Adina , Anagnostopoulou Aikaterini , Raptis Leda
Published: May 2007

To investigate the functional relationship between the ability of the simian virus 40 large tumor antigen (TAg) to transform and its ability to block adipocytic differentiation and induce apoptosis, we expressed TAg in C3H10T1/2 (10T1/2)-derived preadipocytes. The results demonstrated that differentiation could be suppressed at lower TAg levels than at the levels required for full neoplastic conversion. Progressively higher TAg levels were accompanied by apoptosis induction in this system. To further examine the role of the cellular Ras protooncogene product (Ras) in TAg function, TAg was expressed in 10T1/2-derived preadipocytes rendered deficient in Ras activity by transfection with inducible or constitutive antisense ras gene constructs. The results indicated that Ras is required for TAg-mediated transformation and for suppression of adipocytic differentiation, while TAg-mediated apoptosis following serum starvation was independent from Ras action. Unexpectedly, our results further demonstrated a dramatic reduction in the levels of the TAg protein itself as differentiation progressed in Ras-knockdown cells, with a concomitant reduction in TAg’s ability to induce apoptosis as a result. These findings suggest that Ras, although cytoplasmic, is an integral component of the pathway whereby TAg, an oncoprotein believed to have primarily nuclear targets, suppresses differentiation or induces neoplastic conversion of murine preadipocytes.
Effect of cAMP-dependent protein kinase A (PKA) on HCV nucleocapsid assembly and degradation
Authors: Majeau Nathalie , Bolduc Marilène , Duvignaud Jean-Baptiste , Fromentin Rémi , Leclerc Denis
Published: May 2007

The primary function of the hepatitis C virus (HCV) core protein is genome encapsidation. Core protein is also subject to post-translational modifications that can impact on the assembly process. In this report, we have studied the effect of cAMP-dependent protein kinase A (PKA) phosphorylation on its assembly and stability in a yeast Pichia pastoris expression system. We have recently shown that co-expression of the human signal peptide peptidase and core protein (amino acids 1–191) in yeast leads to the formation of nucleocapsid-like particles (NLPs) that are morphologically similar to the wild-type HCV capsid. In this system, we expressed mutants S53A and S116A and mutants S53D and S116D to abolish or mimic PKA phosphorylation, respectively. None of these mutations affected HCV assembly, but S116D led to the degradation of core protein. We also showed that nonenveloped NLPs were labelled in vitro by PKA, suggesting that the phosphorylation sites are available at the surface of the NLPs. The co-expression of human PKA with core and human signal peptide peptidase in yeast did not produce phosphorylated NLPs and led to a decreased accumulation of nonenveloped particles. Mutation S116A restored the core protein content. These results suggest that PKA phosphorylation can modulate HCV core levels in infected cells.
Cloning and characterization of an alternative splicing transcript of the gene coding for human cytidine deaminase
Authors: Lisboa Bianca Cristina Garcia , Machado Tamara da Rocha , Pimenta Daniel Carvalho , Won Han Sang
Published: May 2007

Human cytidine deaminase (HCD) catalyzes the deamination of cytidine or deoxycytidine to uridine or deoxyuridine, respectively. The genomic sequence of HCD is formed by 31 kb with 4 exons and several alternative splicing signals, but an alternative form of HCD has yet to be reported. Here we describe the cloning and characterization of a small form of HCD, HSCD, and it is likely to be a product of alternative splicing of HCD. The alignment of DNA sequences shows that the HSCD matches HCD in 2 parts, except for a deletion of 170 bp. Based on the HCD genome organization, exons 1 and 4 should be joined and all sequences of introns and exons 2 and 3 should be deleted by splicing. This alternative splicing shifted the translation of the reading frame from the point of splicing. The estimated molecular mass is 9.8 kDa, and this value was confirmed by Western blot and mass spectroscopy after expressing the gene fused with glutathionine-S-transferase in the pGEX vector. The deletion and shift of the reading frame caused a loss of HCD activity, which was confirmed by enzyme assay and also with NIH3T3 cells modified to express HSCD and challenged against cytosine arabinoside. In this work we describe the identification and characterization of HSCD, which is the product of alternative splicing of the HCD gene.
The association of HLA-DRB1 alleles with rheumatoid arthritis in the Chinese Shantou population: a follow-up studyThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Lin Ling , Chen Yongsong , Xiao Zhengyu , Huang Shaobi , Yang Ze
Published: May 2007

We investigated the distribution of HLA-DRB1 alleles in a sample of the Chinese Shantou population, and explored the relationship between HLA-DRB1 alleles and the susceptibility and clinical features of rheumatoid arthritis (RA). We studied 117 consecutive patients with RA and control groups, including 38 cases of systemic lupus erythematosus and 100 healthy individuals. HLA-DRB1 genotyping was performed using PCR with sequence-specific primers. HLA-DRB1*04 subtypes were detected using spot hybridization of PCR products with sequence-specific oligonucleotide probes. We compared the frequency of HLA-DRB1 alleles in healthy control patients with that in patients with RA. Patients with RA were evaluated for sex, age at disease onset, disease duration, extra-articular involvement, presence of autoantibodies, global functional status, and radiographic damage. The frequency of HLA-DRB1*04 was found to be significantly higher in RA patients than in healthy individuals (49.6% vs 18.0%, odds ratio = 4.478, P < 0.001). HLA-DRB1*0405 was the most prominently associated subtype in RA patients (62.1% vs 27.8%, odds ratio = 4.255, P = 0.011). Compared with the HLA-DRB1*04-negative RA group, the mean duration of RA in the HLA-DRB1*04-positive RA group was longer, and the mean age at disease onset was lower. A 2–9 year follow-up study was performed, and the risk factors associated with the radiographic progression of RA were determined. Logistic regression analysis revealed that only HLA-DRB1*04 alleles were significantly associated with the radiographic progression of RA (B = 2.652, P = 0.018, Exp(B) = 14.182). Our observations indicated that the HLA-DRB1*04 alleles, especially the subtype HLA-DRB1*0405, were significantly associated with RA susceptibility in the Chinese Shantou population. The HLA-DRB1*04 alleles may be associated with the severity of RA.
The effect of locally deliveredc-mycantisense oligonucleotides combined with intravascular brachytherapy of188Re liquid-filled balloon therapy on vascular smooth muscle cell proliferation in rabbit iliac arteries after injury with a balloon catheterThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Wang Wei , Li Ji-lin , Wang Xin
Published: May 2007

Poststenting restenosis is a significant clinical problem that involves vascular smooth muscle cell (VSMC) proliferation. We primarily investigated the effect of c-myc antisense oligodeoxynucleotides (ASODNs) combined with188Re radiation therapy on VSMC proliferation in rabbit common iliac arteries injured by the porous balloon catheter to explore the therapeutic potential of the combined therapy for the prevention of restenosis. The iliac arteries in rabbits were injured with balloon catheters, and radiation therapy was carried out with a 2.5 mm balloon catheter filled with188Re (8 or 15 Gy), and ASODNs (300 µg) were applied to the adventitia introduced using a pluronic gel releasing system and a lipofectin delivery system. After 3 weeks, the animals were killed and defined segments of arteries were sectioned. The histological sections were stained using α-actin immunohistochemistry staining. The positive α-actin ratios were calculated and analyzed statistically among groups. In contrast to the rate of α-actin positive cell staining in the control group, the rate of α-actin positive cell staining did not decrease (P > 0.05) in the188Re-irradiated group (8 Gy). However, in the ASODN-treated group, the188Re-irradiated group (15 Gy), and the combined ASODN –188Reirradiated groups (8 or 15 Gy), the ratios had markedly decreased (P < 0.01). The effect of the combined group (ASODNs +188Re (15 Gy)) provided the lowest level of α-actin positive cell staining (P < 0.01). The ASODNs (300 µg) effectively decreased VSMC poliferation. The effect of the188Re radiation on the VSMCs depended on the dosage. The ASODNs (300 µg) and combined188Re irradiation effectively lowered VSMC proliferation, and the effect was better than that achieved with any other treatment.
The ASP receptor C5L2 is regulated by metabolic hormones associated with insulin resistance
Authors: MacLaren R. , Kalant D. , Cianflone K.
Published: May 2007

Acylation-stimulating protein (ASP) and interaction with its receptor C5L2 influences adipocyte metabolism. We examined insulin resistance and differentiation-mediated regulation of C5L2 and the mechanistic impact on both C5L2 cell-surface protein and ligand binding to the receptor. C5L2 mRNA increased 8.7-fold with differentiation in 3T3-L1 cells (p < 0.0001) by day 9. In preadipocytes, insulin and dexamethasone increased C5L2 mRNA (1 μmol/L insulin resulted in a 2.6-fold increase, p < 0.01; 10 nmol/L dexamethasone resulted in a 17.9-fold increase, p < 0.01) and C5L2 cell-surface protein (100 nmol insulin resulted in a 2.7-fold increase, p < 0.001; 10 nmol/L dexamethasone resulted in a 2.8-fold increase, p < 0.001). In adipocytes, 100 nmol/L insulin increased C5L2 mRNA and ASP binding (respectively, 1.3-fold, p < 0.01; and 2.4-fold, p < 0.05). Dexamethasone decreased ligand binding (–60%, p < 0.02) without changing mRNA. Tumor necrosis factor alpha decreased C5L2 mRNA (–88% in preadipocytes and –38% in adipocytes, p < 0.001), C5L2 cell-surface protein (–53% in preadipocytes, p < 0.0001), and ASP binding (–60% and –49% in, respectively, preadipocytes and adipoctyes, p < 0.05). Conversely, 1 μmol/L and 10 nmol/L rosiglitazone increased, respectively, C5L2 mRNA (9.3-fold, p < 0.0001) and ASP binding (2.4-fold, p < 0.05). Thus, C5L2 mRNA increases with differentiation, insulin, and thiazolidinedione treatment, and decreases with tumor necrosis factor alpha, all of which results in functional changes in ASP–C5L2 response and may have implications for human metabolism.
Inhibition of osteopontin would suppress angiogenesis in gastric cancer
Authors: Tang Hongwei , Wang Jun , Bai Feihu , Hong Liu , Liang Jie , Gao Juan , Zhai Huihong , Lan Mei , Zhang Faming , Wu Kaichun , Fan Daiming
Published: May 2007

Osteopontin (OPN) plays an important role in tumorigenesis, tumor invasion, and metastasis in many types of cancers, including gastric cancer. Recently, much interest has been focused on the role of OPN in tumor angiogenesis. Our previous studies have shown that OPN is overexpressed, and associated with mean microvessel density in, the tissue samples of patients with gastric cancer. In the present study, we aimed to further determine and provide evidence for the role of OPN in gastric-cancer-associated angiogenesis by diminishing OPN expression in gastric cancer cells using the small interference RNA method, and then evaluate the effects of OPN on gastric cancer-associated angiogenesis by in vivo and in vitro assays. Our results revealed that reduced OPN production by gastric cancer cells would reduce the proliferation, migration, and tube formation of human umbilical vein endothelial cells, and lead to a lower microvessel density, i.e., angiogenesis, in transplanted tumors of mice. These data confirm the positive role of OPN in gastric-cancer-associated angiogenesis.
Staurosporine inhibition of zipper-interacting protein kinase contractile effects in gastrointestinal smooth muscle
Authors: Borman Meredith A. , MacDonald Justin A. , Haystead Timothy A. J.
Published: May 2007

Zipper-interacting protein kinase (ZIPK) is a serine–threonine kinase that has been implicated in Ca2+-independent myosin II phosphorylation and contractile force generation in vascular smooth muscle. However, relatively little is known about the contribution of this kinase to gastrointestinal smooth muscle contraction. The addition of a recombinant version of ZIPK that lacked the leucine zipper domain to permeabilized ileal strips evoked a Ca2+-independent contraction and resulted in myosin regulatory light chain diphosphorylation at Ser19and Thr18. Neither Ca2+-independent force development nor myosin regulatory light chain phosphorylation was elicited by the addition of kinase-dead ZIPK to the ileal strips. The sensitivity of ZIPK-induced contraction to various kinase inhibitors was similar to the in vitro sensitivity of purified ZIPK to these inhibitors. Staurosporine was the most effective ZIPK inhibitor, with a Kivalue calculated to be 2.6 ± 0.3 µmol/L. Through the use of specific kinase inhibitors, we determined that Rho-associated protein kinase and Ca2+/phospholipid-dependent protein kinase (protein kinase C) do not mitigate ZIPK-induced contraction in ileum. Our findings support a role for ZIPK in Ca2+-independent contractile force generation in gastrointestinal smooth muscle.
Cardiovascular diseases in ChinaThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Liu Lisheng
Published: May 2007

Statistics from the National Population Census of China revealed a significant increase in the Chinese population, from 590 million in 1953 to 1.26 billion in 2000. The average life expectancy increased to 71.4 years in 2000 compared with the expectancy of 68.6 years a decade before. World Health Organization statistics on the death rate for total cardiovascular disease, coronary heart disease, and stroke in men and women aged 35–74 years revealed discrepancies between rural and urban parts of China. The China Multicenter Collaborative Study of Cardiovascular Epidemiology indicated that cardiovascular disease was the major cause of death for both men and women, with stroke accounting for over 40% of deaths. Ischemia was shown to be the most common subtype of stroke in both sexes. Smoking was an independent risk factor for cardiovascular disease. The World Health Organization reported that the death rate attributable to tobacco was 6.0% worldwide and 9.2% in China in 1990. The latter is projected to reach 16.6% by 2020. In China, the prevalence of hypertension and diabetes mellitus, the two key risk factors of cardiovascular disease, have also increased significantly in the past 20 years. In addition, elevated blood pressure and plasma cholesterol were two important determinants of increased cardiovascular disease in eastern Asia. These studies indicate that an integrated management of comprehensive risk is urgently required to address China’s increasing cardiovascular disease burden.
Adult asthmatics display exaggerated IFNγ responses to human metapneumovirus and respiratory syncytial virusThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Douville Renée N. , Bastien Nathalie , Li Yan , Simons F. Estelle R. , HayGlass Kent T.
Published: May 2007

Human metapneumovirus and respiratory syncytial virus are RNA viruses associated with lower respiratory tract infections. Regular symptomatic re-infection and sequelae are common, particularly in individuals with pre-existing respiratory diseases, such as asthma. Our understanding of virus-dependent cytokine responses and potential differences between allergic asthmatics and non-asthmatics is limited. To test our hypothesis that adults with mild allergic asthma, the most common form of this disease, exhibit distinct pro-inflammatory responses, we developed a model using acute in vitro infection of fresh peripheral blood mononuclear cells. For both viruses, the production of innate-immunity-associated IL-6 and IL-10 was indistinguishable in the 2 populations. Type 1 cytokine production dominated adaptive immune responses in both asthmatic and non-asthmatic individuals. Surprisingly, asthmatics exhibited stronger pro-inflammatory IFNγ production in response to human metapneumovirus than non-asthmatic adults (p = 0.01), with a similar, but statistically nonsignificant trend in the respiratory-syncytial-virus-stimulated response. Neutralizing IL-10 did not enhance the intensity of IFNγ responses, demonstrating that this pro-inflammatory bias is not counter-regulated by IL-10. Finally, anti-TLR4 blocked lipopolysaccharide, but not respiratory-syncytial-virus-driven cytokine production. Collectively, the data demonstrate that asthma is characterized by markedly stronger pro-inflammatory IFNγ responses to pneumoviruses than their non-asthmatic counterparts. This distinctive pattern of viral immunity may contribute to a worsening of asthma symptoms during respiratory virus infections.
Saikosaponin-d attenuates the development of liver fibrosis by preventing hepatocyte injuryThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Fan Jianghong , Li Xin , Li Ping , Li Ning , Wang Tailing , Shen Hong , Siow Yaw , Choy Patrick , Gong Yuewen
Published: May 2007

Treatment of liver fibrosis and cirrhosis remains a challenging field. Hepatocyte injury and the activation of hepatic stellate cells are the 2 major events in the development of liver fibrosis and cirrhosis. It is known that several Chinese herbs have significant beneficial effects on the liver; therefore, the purpose of the present study was to investigate the therapeutic effect of saikosaponin-d (SSd) on liver fibrosis and cirrhosis. A rat model of liver fibrosis was established using the dimethylnitrosamine method. Liver tissue and serum were used to examine the effect of SSd on liver fibrosis. A hepatocyte culture was also used to investigate how SSd can protect hepatocytes from oxidative injury induced by carbon tetrachloride. The results showed that SSd significantly reduced collagen I deposition in the liver and alanine aminotransferase level in the serum. Moreover, SSd decreased the content of TGF-β1 in the liver, which was significantly elevated after dimethylnitrosamine induced liver fibrosis. Furthermore, SSd was able to alleviate hepatocyte injury from oxidative stress. In conclusion, SSd could postpone the development of liver fibrosis by attenuating hepatocyte injury.
Physiological testosterone stimulates tissue plasminogen activator and tissue factor pathway inhibitor and inhibits plasminogen activator inhibitor type 1 release in endothelial cellsThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Jin Hong , Lin Jijin , Fu Lu , Mei Yi-Fang , Peng Geng , Tan Xuerui , Wang Dong-Ming , Wang Wei , Li Yu-Guang
Published: May 2007

There is a striking gender difference in atherosclerotic vascular disease. For decades, testosterone was considered detrimental to the cardiovascular system. Recent studies, however, have presented some alternative results. The aim of this study was to evaluate the effect of testosterone, using physiological and supraphysiological concentrations, on antigen and mRNA levels of tissue plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI-1), and tissue factor pathway inhibitor (TFPI) released by human umbilical vein endothelial cells and to investigate the cellular mechanism. Cells within 2–3 passages were cultured in 25 cm2 flasks or plated onto 96-well plates with a density of about 1 × 105 cells/mL as recommended. The cells were incubated in the presence or absence of testosterone (3, 30, 3 × 103, 3 × 104 nmol/L) for 48 h. Levels of tPA, PAI-1, and TFPI antigen were assayed with ELISA kits. Reverse transcriptase PCR was carried out to detect tPA, PAI-1, and TFPI mRNA levels. Cells were incubated in androgen-receptor antagonist (flutamide 10 µmol/L) or aromatase inhibitor (aminoglutethimide 50 µmol/L) for 3 h, and then the experiments were repeated. Testosterone at a physiologic concentration (30 nmol/L) increased the antigen levels of tPA and TFPI significantly (P < 0.05). However, tPA and TFPI levels were markedly reduced (P < 0.05) at a larger dose (3 × 104 nmol/L). On the other hand, PAI-1 antigen levels decreased significantly at the testosterone concentrations ranging from 3 to 3 × 104 nmol/L (P < 0.05). The change in the levels of tPA and TFPI were reflected in the corresponding change in mRNA levels. Flutamide attenuated the effect of testosterone at physiological concentration (30 nmol/L). The results demonstrated that testosterone at physiological concentrations may have a beneficial influence on the haemostatic system through enhancement of anticoagulant activity, resulting from stimulation of TFPI and tPA expression and inhibition of PAI-1 secretion by the endothelium.
Effects ofN-n-butyl haloperidol iodide on L-type calcium channels and intracellular free calcium in rat ventricular myocytesThis paper is one of a selection of papers in this Special Issue, entitled International Symposium on Recent Advances in Molecular, Clinical, and Social Medicine, and has undergone the Journal's usual peer-review process.
Authors: Huang Zhanqin , Shi Ganggang , Gao Fenfei , Zhang Yanmei , Liu Xingping , Christopher Theodore A. , Lopez Bernard , Ma XinLiang
Published: May 2007

The ability of N-n-butyl haloperidol iodide (F2) to cause vasodilation, and thereby produce a cardioprotective effect, has been well documented. The aim of this study was to investigate whether F2might act as a Ca2+antagonist. Myocytes were obtained from rat heart, and the whole-cell patch-clamp technique was used to record Ca2+current. Laser scanning confocal microscopy was used to measure intracellular free calcium ([Ca2+]i). The results obtained from this study demonstrate that F2reduced calcium current (ICa) in a concentration-dependent manner with an IC50of 1.19 µmol/L, upshifted the current-voltage curve of ICa, shifted the inactivation kinetics of ICaleftward, and slowed down the recovery of ICafrom inactivation. F2decreased the fluorescent intensity of [Ca2+]ielevation induced by KCl with an IC50of 1.61 µmol/L, and had no effects on the intracellular calcium release induced by caffeine and inositol-1,4,5-trisphosphate. These findings indicate that F2may act as a calcium antagonist, which could account for its cardiovascular benefits.