bluuebunny/crossref_metadata_2025 huggingface dataset:
Features that determine telomere homolog oligonucleotide‐induced therapeutic DNA damage‐like responses in cancer cells
Authors: Ohashi Norio , Yaar Mina , Eller Mark S. , Truzzi Francesca , Gilchrest Barbara A.
Published: November 2006

Cancer is the second leading cause of death in the USA, with metastatic disease proving a particular management challenge. Treatment modalities for patients with metastatic disease are limited, and survival beyond 5 years is uncommon. We have reported that an 11‐base DNA oligonucleotide 100% homologous to the telomere 3′ overhang can induce apoptosis, senescence and/or differentiation of several types of malignant cells in vitro and in vivo, while having minimal effect on normal cells. We now report that 22 oligonucleotides, 9–20 bases in length, with or without a 5′ phosphate group and with varying homology (40–100%) to the 3′ overhang, inhibit growth and induce apoptosis of human cell lines derived from breast cancers, pancreatic and ovarian carcinomas, and malignant melanoma, lines that lack p53 and/or p16 and harbor a variety of other abnormalities in key regulatory signaling pathways. Cytosine (C) content adversely affected oligonucleotide efficacy, decreasing their effect on cellular apoptosis by ≥80%. These data confirm and expand our earlier work suggesting that such telomere homolog oligonucleotides (T‐oligos) target an innate anti‐cancer defense system in human cells and may provide an effective treatment for cancers of multiple different cellular origins and genetic profile. J. Cell. Physiol. 210: 582–595, 2007. © 2006 Wiley‐Liss, Inc.
IGF‐1 upregulates electroneutral K‐Cl cotransporter KCC3 and KCC4 which are differentially required for breast cancer cell proliferation and invasiveness
Authors: Hsu Yueh‐Mei , Chou Cheng‐Yang , Chen Helen H.W. , Lee Wen‐Ying , Chen Yih‐Fung , Lin Pin‐Wen , Alper Seth L. , Ellory J. Clive , Shen Meng‐Ru
Published: November 2006

The cellular function of electroneutral K‐Cl cotransport (KCC) is to regulate epithelial ion transport and osmotic homeostasis. Here we investigate the mechanisms by which insulin‐like growth factor 1 (IGF‐1) cooperates with KCC to modulate breast cancer biology. IGF‐1 stimulates KCC activity of MCF‐7 breast cancer cells in a dose‐ and time‐dependent manner. Increased KCC3 and KCC4 abundances contribute to IGF‐1‐enhanced KCC activity. Endogenous cellular invasiveness was modestly attenuated by KCC4‐specific siRNA and the residual invasiveness was much less sensitive to IGF‐1 stimulation. KCC3 knockdown significantly reduced basal growth rate and almost abolished IGF‐1‐stimulated cell proliferation. Consistently, MCF‐7 cells obtained advantage in cell proliferation and invasiveness by overexpression of KCC3 and KCC4, respectively. Blockade of gene transcription by actinomycin D abolished IGF‐1‐mediated increase in KCC3 and KCC4 mRNA, indicating that IGF‐1 increases KCC abundance through the regulation of KCC genes. IGF‐1 treatment triggered phosphatidylinositol 3‐kinase and mitogen‐activated protein kinase (MAPK) cascades which were differentially required for IGF‐1‐stimulated biosynthesis of KCC3 and KCC4. Loss‐of‐function mutations in KCC significantly inhibited the development and progression of xenograft tumor in SCID mice. The expression level of IGF‐1 and KCC polypeptides in the surgical specimens showed a good linear correlation, suggesting autocrine or paracrine IGF‐1 stimulation of KCC production in vivo. Among patients with early‐stage node‐negative breast cancer, disease‐free survival (DFS) and overall survival (OS) curves were significantly different based on IGF‐1 and KCC expression. Thus, we conclude that KCC activation by IGF‐1 plays an important role in IGF‐1 receptor signaling to promote growth and spread of breast cancer cells. J. Cell. Physiol. 210: 626–636, 2007. © 2006 Wiley‐Liss, Inc.
Potent stimulation of large‐conductance Ca2+‐activated K+ channels by rottlerin, an inhibitor of protein kinase C‐δ, in pituitary tumor (GH3) cells and in cortical neuronal (HCN‐1A) cells
Authors: Wu Sheng‐Nan , Wang Ya‐Jean , Lin Ming‐Wei
Published: November 2006

The effects of rottlerin, a known inhibitor of protein kinase C‐δ activation, on ion currents were investigated in pituitary tumor (GH3) cells. Rottlerin (0.3–100 µM) increased the amplitude of Ca2+‐activated K+ current (IK(Ca)) in a concentration‐dependent manner with an EC50 value of 1.7 µM. In intracellular perfusion with rottlerin (1 µM) or staurosporine (10 µM), phorbol 12‐myristate 13‐acetate‐induced inhibition of IK(Ca) in these cells was abolished. In cell‐attached mode, rottlerin applied on the extracellular side of the membrane caused activation of large‐conductance Ca2+‐activated K+ (BKCa) channels, and a further application of BAPTA‐AM (10 µM) to the bath had no effect on rottlerin‐stimulated channel activity. When cells were exposed to rottlerin, the activation curve of these channels was shifted to less positive potential with no change in the slope factor. Rottlerin increased BKCa‐channel activity in outside‐out patches. Its change in kinetic behavior of BKCa channels is primarily due to an increase in mean open time. With the aid of minimal kinetic scheme, a quantitative description of rottlerin stimulation on BKCa channels in GH3 cells was also provided. Under current‐clamp configuration, rottlerin (1 µM) decreased the firing of action potentials. IK(Ca) elicited by simulated action potential waveforms was enhanced by this compound. In human cortical HCN‐1A cells, rottlerin (1 µM) could also interact with the BKCa channel to stimulate IK(Ca). Therefore, rottlerin may directly activate BKCa channels in neurons or endocrine cells. J. Cell. Physiol. 210: 655–666, 2007. © 2006 Wiley‐Liss, Inc.
Multiple binding of methyl‐CpG and polycomb proteins in long‐term gene silencing events
Authors: Matarazzo M.R. , De Bonis M.L. , Strazzullo M. , Cerase A. , Ferraro M. , Vastarelli P. , Ballestar E. , Esteller M. , Kudo S. , D'Esposito M.
Published: November 2006

Epigenetic regulation is involved in the maintenance of long‐term silencing phenomena, such as X‐inactivation and genomic imprinting in mammals. Gene repression is mediated by several mechanisms, such as histone modifications, DNA methylation, and recruitment of Polycomb proteins. To understand the mechanistic relationships between these mechanisms for stable gene silencing, we analyzed the mechanisms of X‐ and Y‐inactivation of the PAR2 gene SYBL1, previously showed to be regulated by concerted epigenetic mechanisms. Maintenance of stable repression occurs via the recruitment of both MBDPs and PRC2 complexes to SYBL1 promoter. Their binding is equally sensitive to defective DNA methylation seen in cells derived from ICF syndrome patients. Multiple occupancy is a feature shared within long‐term repressed genes, such as the X‐inactivated PGK1 and the imprinted IGF2. MBD2, MBD3, and MeCP2 occupy SYBL1 promoter simultaneously, as revealed by sequential ChIP. We did not find this co‐occurring binding when looked for members of PRC2 complex together with any of the methyl‐binding proteins. Furthermore, in co‐transfection assays, MECP2 can silence methylated SYBL1 promoter, whereas the mutated protein fails. However, RNA interference of endogenous MECP2 does not induce the expression of the inactive SYBL1 alleles, suggesting that its silencing activity can be replaced by the other methyl‐binding proteins. Our data suggest that maintenance of long‐term silencing involves multiple layers of epigenetic control functionally redundant. PRC2 and MBD proteins could collaborate to different phases of this process, the former possibly recruiting DNMTs to the silenced promoters, the latter dictating the lock of the transcription. J. Cell. Physiol. 210: 711–719, 2007. © 2006 Wiley‐Liss, Inc.
MAP kinase‐dependent, NF‐κB‐independent regulation of inhibitor of apoptosis protein genes by TNF‐α
Authors: Furusu Akira , Nakayama Kenji , Xu Qihe , Konta Tsuneo , Kitamura Masanori
Published: November 2006

The inhibitor of apoptosis protein (IAP) family of molecules regulates apoptotic processes triggered by various stimuli. However, the mechanisms involved in the regulation of the IAP genes are not fully understood. In this report, we examined roles of nuclear factor‐κB (NF‐κB) and mitogen‐activated protein (MAP) kinases in tumor necrosis factor‐α (TNF‐α)‐induced expression of IAP genes. In human endothelial cells, TNF‐α induced c‐IAP1 and c‐IAP2, but not XIAP and TIAP/Survivin, at the transcriptional level. Inactivation of NF‐κB by overexpression of a super‐repressor mutant of IκBα did not affect the induction of IAPs by TNF‐α. In contrast, extracellular signal‐regulated kinase, p38 MAP kinase, and c‐Jun N‐terminal kinase were activated after stimulation with TNF‐α, and inhibition of each kinase by PD098059, SB203580, curcumin, or SP600125 substantially attenuated the TNF‐α‐induced c‐IAP1 and c‐IAP2 expression. To examine whether the MAP kinases‐mediated induction of IAPs contributes to survival of TNF‐α‐exposed cells, cells were pretreated with MAP kinase inhibitors and stimulated with TNF‐α. Treatment with kinase inhibitors alone did not induce apoptosis but enhanced markedly TNF‐α‐triggered apoptosis. Furthermore, overexpression of either c‐IAP1 or c‐IAP2 diminished the apoptosis‐promoting effects of MAP kinase inhibitors. These data indicated that TNF‐α induced expression of c‐IAP1 and c‐IAP2 via MAP kinases, but not via NF‐κB, and that MAP kinases participated in the inhibition of apoptosis by induction of c‐IAPs in TNF‐α‐stimulated endothelial cells. J. Cell. Physiol. 210: 703–710, 2007. © 2006 Wiley‐Liss, Inc.
Molecular targets for retinal vascular diseases
Authors: Campochiaro Peter A.
Published: November 2006

The elucidation of the molecular pathogenesis of a disease in animal models provides candidate targets for treatment. As specific antagonists for a target are developed and tested in clinical trials, if benefit is achieved, the candidate becomes a validated target. Validated targets stimulate additional research to identify optimal ways of attacking the target and studies in related disease processes to determine if the molecule is also a target in that context. Vascular endothelial growth factor (VEGF) has been identified as a validated target for several retinal vascular diseases. This has led to a flurry of activity resulting in beneficial treatments for patients and intensification of the search for other targets. This review summarizes preclinical and clinical trial results obtained with VEGF antagonists and describes evidence supporting the candidacy of other molecules currently being tested or soon to be tested for target status. J. Cell. Physiol. 210: 575–581, 2007. © 2006 Wiley‐Liss, Inc.
Oncogenic KRAS provides a uniquely powerful and variable oncogenic contribution among RAS family members in the colonic epithelium
Authors: Keller Jeffrey W. , Franklin Jeffrey L. , Graves‐Deal Ramona , Friedman David B. , Whitwell Corbin W. , Coffey Robert J.
Published: November 2006

Activating mutations of the RAS family of small GTPases are among the most common genetic events in human tumorigenesis. Constitutive activation of the three canonical family members, KRAS, NRAS, and HRAS segregate strongly by tissue type. Of these, KRAS mutations predominate in human tumors, including those arising from the colon and lung. We sought to compare the oncogenic contributions of different RAS isoforms in a comparable genetic setting and to explore downstream molecular changes that may explain the apparent differential oncogenic effects of the various RAS family members. We utilized colorectal cancer cell lines characterized by oncogenic KRAS in parallel with isogenically derived lines in which the mutant allele has been disrupted. We additionally attempted to reconstitute the isogenic derivatives with oncogenic forms of other RAS family members and analyze them in parallel. Pairwise analysis of HCT 116 and DLD‐1 cell lines as well as their isogenic derivatives reveals distinct K‐RASG13D signatures despite the genetic similarities of these cell lines. In DLD‐1, for example, oncogenic K‐RAS enhances the motility of these cells by downregulation of Rap1 activity, yet is not associated with increased ERK1/2 phosphorylation. In HCT 116, however, ERK1/2 phosphorylation is elevated relative to the isogenic derivative, but Rap1 activity is unchanged. K‐RAS is uniquely oncogenic in the colonic epithelium, though the molecular aspects of its oncogenic contribution are not necessarily conserved across cell lines. We therefore conclude that the oncogenic contribution of K‐RAS is a function of its multifaceted functionality and is highly context‐dependent. J. Cell. Physiol. 210: 740–749, 2007. © 2006 Wiley‐Liss, Inc.
Role of atypical protein kinase C isozymes and NF‐κB in IL‐1β‐induced expression of cyclooxygenase‐2 in human myometrial smooth muscle cells
Authors: Duggan Sara V. , Lindstrom Tamsin , Iglesias Teresa , Bennett Phillip R. , Mann Giovanni E. , Bartlett Simon R.
Published: November 2006

Increased myometrial expression of cyclooxygenase‐2 (Cox‐2) at term results from elevated local levels of inflammatory cytokines, and its inhibition provides a potential route for intervention in human pre‐term labor. We have identified a role for atypical protein kinase C (PKC) isozymes in IL‐1β‐induced Cox‐2 expression in human myometrial smooth muscle cells (HMSMC). The PKC inhibitor GF109203X (10 µM) inhibited IL‐1β‐induced Cox‐2 protein and RNA expression, which were also reduced by MAPK and nuclear factor κB (NF‐κB) inhibitors. GF109203X did not affect MAPK activities, and neither did it replicate the effect of p38 MAPK inhibition on Cox‐2 mRNA stability, suggesting that PKC operates through an independent mechanism. The effect of GF109203X remained intact after depletion of conventional and novel PKC isozymes by phorbol ester pre‐treatment. In contrast LY379196 (10 µM), which at micromolar concentrations inhibits all but atypical PKCs, did not affect Cox‐2 expression. A peptide corresponding to the pseudosubstrate sequence of atypical PKCs blocked Cox‐2 protein expression, whereas the sequence from conventional PKCs was ineffective. GF109203X did not affect NF‐κB binding to nuclear proteins, but strongly reduced NF‐κB‐dependent transcription in luciferase reporter assays. Our findings indicate that IL‐1β‐induced Cox‐2 expression in HMSMC in culture requires p38‐MAPK‐mediated mRNA stabilization and an independent activation of Cox‐2 transcription which is dependent on the action of atypical PKCs, probably through direct stimulation of the transactivating activity of NF‐κB. J. Cell. Physiol. 210: 637–643, 2007. © 2006 Wiley‐Liss, Inc.
CCL20 chemokine induces both osteoblast proliferation and osteoclast differentiation: Increased levels of CCL20 are expressed in subchondral bone tissue of rheumatoid arthritis patients
Authors: Lisignoli Gina , Piacentini Anna , Cristino Sandra , Grassi Francesco , Cavallo Carola , Cattini Luca , Tonnarelli Beatrice , Manferdini Cristina , Facchini Andrea
Published: November 2006

We evaluated the role of CCL20 (MIP‐3α) chemokine in cells directly involved in the remodeling of bone tissue (osteoblasts and osteoclasts) and we confirmed its expression in the subchondral bone tissue of rheumatoid arthritis (RA) patients. The expression of CCL20 and of its receptor CCR6 was evaluated in osteoblasts isolated from bone tissue of post‐traumatic (PT) patients. Functional tests were performed to evaluate osteoblast proliferation and matrix protein modulation. Immunohistochemical analysis for CCR6, CCL20, and RANKL was performed on bone samples from RA patients. The role of CCL20 was then analyzed in osteoclast differentiation. We found that in basal conditions CCR6, but not its ligand CCL20, was highly expressed by osteoblasts. Functional analysis on osteoblasts showed that CCL20 significantly increased cellular proliferation but did not affect matrix protein expression. Pro‐inflammatory cytokines significantly induced the release of CCL20 and RANKL by human osteoblasts but did not modulate CCR6 expression. Increased expression of CCR6, CCL20, and RANKL was confirmed in RA subchondral bone tissue biopsies. We demonstrated that CCL20 was also an earlier inducer of osteoclast differentiation by increasing the number of pre‐osteoclasts, thus favoring cell fusion and MMP‐9 release. Our results add new insight to the important role of the CCL20/CCR6, RANKL system in the bone tissue of RA. The contemporary action of CCL20 on osteoblasts and osteoclasts involved in the maintenance of bone tissue homeostasis demonstrates the important role of this compartment in the evolution of RA, by showing a clear uncoupling between new bone formation and bone resorption. J. Cell. Physiol. 210: 798–806, 2007. © 2006 Wiley‐Liss, Inc.
Molecular signaling and genetic pathways of senescence: Its role in tumorigenesis and aging
Authors: Zhang Hong
Published: November 2006

In response to progressive telomere shortening in successive cell divisions, normal somatic cells enter senescence, during which they cease to proliferate irreversibly and undergo dramatic changes in gene expression. Senescence can also be activated by various types of stressful stimuli, including aberrant oncogenic signaling, oxidative stress, and DNA damage. Because of the limited proliferative capacity imposed by senescence, as well as the ability of senescent cells to influence neighboring non‐senescent cells, senescence has been proposed to play an important role in tumorigenesis and to contribute to aging. Considerable effort has been put into elucidating the molecular mechanisms of senescence, including the signals that trigger senescence, the molecular pathways by which cells enter senescence, and evidence that supports its role in tumorigenesis and aging. J. Cell. Physiol. 210: 567–574, 2007. © 2006 Wiley‐Liss, Inc.
2,2′‐Thiodiethanol: A new water soluble mounting medium for high resolution optical microscopy
Authors: Staudt Thorsten , Lang Marion C. , Medda Rebecca , Engelhardt Johann , Hell Stefan W.
Published: November 2006

The use of high numerical aperture immersion lenses in optical microscopy is compromised by spherical aberrations induced by the refractive index mismatch between the immersion system and the embedding medium of the sample. Especially when imaging >10 μm deep into the specimen, the refractive index mismatch results in a noticeable loss of image brightness and resolution. A solution to this problem is to adapt the index of the embedding medium to that of the immersion system. Unfortunately, not many mounting media are known that are both index tunable as well as compatible with fluorescence imaging. Here we introduce a nontoxic embedding medium, 2,2′‐thiodiethanol (TDE), which, by being miscible with water at any ratio, allows fine adjustment of the average refractive index of the sample ranging from that of water (1.33) to that of immersion oil (1.52). TDE thus enables high resolution imaging deep inside fixed specimens with objective lenses of the highest available aperture angles and has the potential to render glycerol embedding redundant. The refractive index changes due to larger cellular structures, such as nuclei, are largely compensated. Additionally, as an antioxidant, TDE preserves the fluorescence quantum yield of most of the fluorophores. We present the optical and chemical properties of this new medium as well as its application to a variety of differently stained cells and cellular substructures. substructures. Microsc. Res. Tech., 2007. © 2006 Wiley‐Liss, Inc.
A branched histidine/lysine peptide, H2K4b, in complex with plasmids encoding antitumor proteins inhibits tumor xenografts
Authors: Leng Qixin , Scaria Puthupparampil , Ioffe Olga B. , Woodle Martin , Mixson A. James
Published: November 2006

Background

In this study we investigated whether a particular branched HK polymer, H2K4b, was an effective in vivo carrier of plasmids expressing the antiangiogenic kringle 1–5 or the tumor suppressor p53.

Methods

H2K4b was synthesized on a solid‐phase peptide synthesizer. Distribution, optimization and time course studies were done in tumor‐bearing nude mice by systemically administering H2K4b in complex with a luciferase‐expressing plasmid. We examined the amount of tumor angiogenesis in C6 with MDA‐MB‐435 xenografts utilizing the carmine dye. The ability of H2K4b to carry luciferase plasmids to different tissues was compared with several liposomal carriers. Medium from cells transfected with mKr1‐5 was tested for its capacity to inhibit angiogenesis with an in vivo Matrigel assay. We then determined if systemically delivered H2K4b in complex with plasmid encoding mKr1‐5 inhibited tumor growth; we also compared the antitumor activity of HK polyplexes containing hKr1‐5, mKr1‐5, and p53 plasmids.

Results

H2K4b carried the luciferase‐expressing plasmid in order of descending efficacy to these tissues: lung, spleen, tumor, and liver. Compared to DOTAP‐containing liposomes, H2K4b was a more effective carrier of a luciferase‐containing plasmid to extrapulmonary tissues. We then determined that mKr1‐5 in complex with H2K4b reduced MDA‐MB‐435 tumor growth by approximately 50% compared to the control group (P < 0.01). Similarly, H2K4b/mKr1‐5 polyplexes reduced the growth of C6 xenografts. In MDA‐MB‐435 xenografts, p53‐ and Kr1‐5‐expressing plasmids in complex with H2K4b had comparable antitumor activity.

Conclusion

H2K4b demonstrates potential as a carrier of plasmids encoding antiangiogenic and/or tumor suppressor proteins in a tumor‐bearing mouse model. Copyright © 2006 John Wiley & Sons, Ltd.
Social capital, egalitarianism and foreign aid allocations
Authors: Knowles Stephen
Published: November 2006

This paper explores the issue of whether countries that have higher levels of social capital, and/or are more egalitarian, are more generous in terms of donating foreign aid. The empirical results suggest that in countries with a more equal distribution of income, aid allocations by the government are higher, but donations to non‐government aid organisations by the private sector are lower. There is a positive correlation between the level of social capital and aid allocated by both the government and the private sector. Copyright © 2006 John Wiley & Sons, Ltd.
The nature of indigenous environmental knowledge production: evidence from Bedouin communities in southern Egypt
Authors: Briggs John , Sharp Joanne , Yacoub Hoda , Hamed Nabila , Roe Alan
Published: November 2006

The use of indigenous knowledge has been seen in some quarters to offer real possibilities of success in development practice. However, results have been uneven, perhaps because of the way in which indigenous knowledge has been conceptualised. Drawing on empirical research among two related Bedouin communities in Egypt, the paper suggests that indigenous knowledge is provisional and dynamic and therefore rather less static than implied in much of the literature; it should be seen as utilitarian and grounded, both economically and socio‐culturally; and indigenous knowledge as a term may be unhelpful and misleading and would be better expressed as local knowledges. Copyright © 2006 John Wiley & Sons, Ltd.
Rapid immunochromatographic test for hantavirus andes contrasted with capture‐IgM ELISA for detection of andes‐specific IgM antibodies
Authors: Navarrete Maritza , Barrera Claudia , Zaror Luis , Otth Carola
Published: November 2006

Hantavirus is associated with hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). The clinical diagnosis of hantavirus infections has been confirmed routinely by the immunofluorescence antibody assay (IFA) or enzyme‐linked immunosorbent assay (ELISA). A rapid and easy diagnostic test for hantavirus infection is required. A new immunochromatographic assay for hantavirus, POC‐PUU, useful for the diagnosis of epidemic nephrophaty associated with hantavirus Puumala in Europe, was evaluated in Chile. This test is based on recombinant N‐protein of hantavirus Puumala, and cross‐reacts with other hantaviruses. Eighty human sera were selected at random from patients from Southern Chile who were suspected with HPS. The hantavirus capture‐IgM ELISA was compared with a commercially available POC‐PUU test (POC PUUMALA, Reagena Ltd., Toivala, Finland). The test sensitivity and specificity of the POC‐PUU test were 97 and 90%, respectively. It is important to note that although the test is not specific for Andes virus the sensitivity and specificity were above 90%, which indicates good reactivity to the Puumala nucleoprotein antigen. As this test is cost‐effective, with a high negative value, rapid and easy to carry out, specialized personnel are not necessary, nor does it require specialized equipment. Its usefulness for diagnosis is important in hospitals far from reference centers and areas with a high incidence of HPS cases. J. Med. Virol. 79:41–44, 2007. © 2006 Wiley‐Liss, Inc.
Differences in replication capacity between enterovirus 71 isolates obtained from patients with encephalitis and those obtained from patients with herpangina in Taiwan
Authors: Kung Chien‐Min , King Chwan‐Chuen , Lee Chun‐Nan , Huang Li‐Min , Lee Ping‐Ing , Kao Chuan‐Liang
Published: November 2006

The cellular‐tropism and biological characteristics of enterovirus 71 (EV71) isolates in Taiwan (TW) were studied. Growth curve experiments were conducted using cell lines that were possibly exhibited pathogenesis, and RT‐PCR and sequencing tests were undertaken to amplify the 5′ non‐coding region (5′‐NCR). The encephalitis isolate EV71 TW98NTU2078 was PBMC‐tropic, temperature‐resistant (Tr) at 40°C, and easier to replicate in HTB‐14 (astrocytoma) than the herpangina isolate EV71 TW98NTU1186 (The viral yields were 100‐fold higher than those of the herpangina isolate EV71 TW98NTU1186 at 96 hr post infection.). The herpangina isolate EV71 TW98NTU1186 was non‐PBMC‐tropic, and temperature‐sensitive (Ts) at 40°C. The replication of EV71 TW98NTU1186 in HTB‐14 was lower. No EV71 isolate infected HTB‐37 (human colon adenocarcinoma cells). The encephalitis EV71 isolate exhibited better replication and transmission in PBMCs and astrocytes than did the EV71 isolate without CNS involvement. J. Med. Virol. 79:60–68, 2007. © 2006 Wiley‐Liss, Inc.
Norovirus capsid protein expressed in yeast forms virus‐like particles and stimulates systemic and mucosal immunity in mice following an oral administration of raw yeast extracts
Authors: Xia Ming , Farkas Tibor , Jiang Xi
Published: November 2006

Norovirus (NV) gastroenteritis is a widespread disease affecting people of all ages worldwide. A simple, safe, and easily deliverable vaccine may be the key for the control and prevention of NV gastroenteritis. In this study, we demonstrated that a NV recombinant capsid protein (strain VA387, genogroup II.4) expressed in yeast (Pichia pastoris) spontaneously formed virus‐like particles (VLPs) like those expressed in other in vitro systems. Oral administration of raw material from the yeast cell lysates containing 0.1 mg of VLPs without an adjuvant resulted in systemic and mucosal immune responses in mice. Significantly higher and earlier responses were observed in mice receiving a higher dose (1 mg per dose) of the antigen. Both the serum and fecal antibodies blocked VA387 VLP binding to its histo‐blood group antigen receptors. The animals did not reveal any side effect following the administration of the yeast lysates. Our results indicated that yeast is a simple, effective alternative for NV VLP production. The mice immunization study also indicated that the oral administration of raw yeast extracts without an adjuvant is a safe and simple way which is worth to be studied for vaccine delivery in humans. J. Med. Virol. 79:74–83, 2007. © 2006 Wiley‐Liss, Inc.
Identification of NV‐F virus DNA in hepatocellular carcinoma
Authors: Yeh Chau‐Ting , Chen Tse‐Ching , Chang Ming‐Ling , Hsu Chao‐Wei , Yeh Ta‐Sen , Lee Wei‐Chen , Huang Shiu‐Feng , Tsai Chih‐Chieh
Published: November 2006

A fragment of DNA sequence derived from a hepatotropic virus, named NV‐F was isolated recently. The aim of this study was to examine whether this virus was associated with hepatocellular carcinoma (HCC). Total cellular DNA was extracted from hepatocellular carcinoma tissues. NV‐F virus DNA was detected by PCR. The PCR products were subjected to sequence analysis. Of the 78 HCC samples included, 12 (15.4%) were positive for NV‐F virus DNA. Sequence analysis of the 12 amplified DNA fragments revealed a point mutation in one of them. The clinicopathological parameters between patients with and without NV‐F virus infection were compared. It was found that patients with NV‐F virus infection were older than those without NV‐F virus infection (mean ages, 61.5 versus 52.5 years; P = 0.032). Otherwise, no difference was observed between the two groups. Of the 12 HCC patients positive for NV‐F virus DNA, 11 patients were co‐infected by either hepatitis B or C virus. The remaining patient was a Taiwanese aboriginal inhabitant with cryptogenic cirrhosis. In conclusion, NV‐F virus DNA was identified in 15.4% of HCC tissues. HCC patients with NV‐F virus infection were significantly older than those without NV‐F virus infection. J. Med. Virol. 79:92–96, 2007. © 2006 Wiley‐Liss, Inc.
Seroprevalence of rubella infection after national immunization program in Taiwan: Vaccination status and immigration impact
Authors: Wang I‐Jen , Huang Li‐Ming , Chen Hsiu‐Hsi , Hwang Kung‐Chang , Chen Chien‐Jen
Published: November 2006

Rubella vaccination was implemented nationwide in 1992 in Taiwan. A cross‐sectional survey was conducted to determine the age‐specific seroprevalence of anti‐rubella in female students aged 7–22 years old and women of reproductive age in Taipei County. Seropositivity of anti‐rubella antibodies was defined as a serum IgG level ≥10 IU/ml tested by enzyme immunoassay. Information on possible predictors of anti‐rubella seronegativity was obtained from structured questionnaire interview. Multivariate‐adjusted odds ratios (ORm) with their 95% confidence interval (CI) were derived for each predictor using multiple logistic regression analysis. A total of 826 female students and 318 women were recruited. Anti‐rubella seropositive rates were 98.3% (282/287), 99.6% (234/235), 96.8% (179/185), 92.4% (110/119), 81.4% (197/242), and 89.5% (68/76) for the age groups of 7–9, 11–13, 15–17, 19–22, 25–33, and 34–44 years old, respectively. Among female students, negative serology for rubella antibodies was associated significantly with the age group and foreign nationality of mother, showing ORm (95% CI) of 1.2 (1.06–1.27) for each year increase in age, and 20.9 (6.31–68.97), respectively. Among women at the reproductive age, low maternal education level 91.6 (9.12–920.74), unmarried status 21.2 (6.16–72.89), and no rubella vaccination 98.9 (11.64–840.25) were associated significantly with an increased evidence of seronegativity to rubella. The National Rubella Vaccination Program has led to herd immunity of school girls. However, in order to eradicate the Rubella syndrome, greater effort is required to vaccinate foreign brides in Taiwan. J. Med. Virol. 79:97–103, 2007. © 2006 Wiley‐Liss, Inc.
Osmotic regulation of neuronal nitric oxide synthase expression in the rat amygdala: Functional role for nitric oxide in adaptive responses?
Authors: Yao Song T. , Antunes Vagner R. , Paton Julian F.R. , Murphy David
Published: November 2006

Water‐deprivation‐induced osmotic stress leads to activation of a number of adaptive responses. Nitric oxide (NO) has been implicated in the modulation of these responses, as the amygdala has been implicated in ingestive behavior and modulation of autonomic homeostatic functions. Here we investigated the effects of water deprivation on neuronal nitric oxide synthase (nNOS) expression within the rat amygdala; a brain area involved in modulating ingestive behavior and autonomic function. Water deprivation resulted in significant increases in nNOS immunoreactivity (‐ir) within different regions of the amygdala compared with euhydrated rats. Maximal increases were observed in the anteroventral (118 ± 9 vs. 47 ± 3 neurons), anteriodorsal (133 ± 9 vs. 77 ± 3), and posterioventral (175 ± 5 vs. 71 ± 5) parts of the medial amygdala. The basomedial nucleus (65 ± 4 vs. 39 ± 3) and posterior basolateral nucleus (19 ± 2 vs. 5 ± 1) of the amygdala and the capsular (21 ± 2 vs. 6 ± 1) and medial (44 ± 6 vs. 22 ± 3) parts of the central nucleus of the amygdala also showed increased nNOS‐ir in dehydrated rats. Water deprivation had no effect on nNOS‐ir in areas such as the cortical, anterior basolateral, and intercalated nuclei of the amygdala. Microinjection of an NO donor, DEA‐NONOate, into the central amygdala resulted in a pressor and tachycardic response that was attenuated by a soluble guanylate cyclase inhibitor. These observations suggest that activation of the nitrergic system is prevalent throughout the amygdala following water deprivation and suggest that the up‐reguation of nNOS could play a significant role in the integrative response to osmotic stress. © 2006 Wiley‐Liss, Inc.
In vitro characterization of a human neural progenitor cell coexpressing SSEA4 and CD133
Authors: Barraud Perrine , Stott Simon , Møllgård Kjeld , Parmar Malin , Björklund Anders
Published: November 2006

The stage‐specific embryonic antigen 4 (SSEA4) is commonly used as a cell surface marker to identify the pluripotent human embryonic stem (ES) cells. Immunohistochemistry on human embryonic central nervous system revealed that SSEA4 is detectable in the early neuroepithelium, and its expression decreases as development proceeds. Flow cytometry analysis of forebrain‐derived cells demonstrated that the SSEA4‐expressing cells are enriched in the neural stem/progenitor cell fraction (CD133+), but are rarely codetected with the neural stem cell (NSC) marker CD15. Using a sphere‐forming assay, we showed that both subfractions CD133+/SSEA4+ and CD133+/CD15+ isolated from the embryonic forebrain are enriched in neurosphere‐initiating cells. In addition CD133, SSEA4, and CD15 expression is sustained in the expanded neurosphere cells and also mark subfractions of neurosphere‐initiating cells. Therefore, we propose that SSEA4 associated with CD133 can be used for both the positive selection and the enrichment of neural stem/progenitor cells from human embryonic forebrain. © 2006 Wiley‐Liss, Inc.
Developmental abnormalities in the nerves of peripheral myelin protein 22‐deficient mice
Authors: Amici Stephanie A. , Dunn William A. , Notterpek Lucia
Published: November 2006

Peripheral myelin protein 22 (PMP22) is a tetraspan glycoprotein whose misexpression is associated with a family of hereditary peripheral neuropathies. In a recent report, we have characterized a novel PMP22‐deficient mouse model in which the first two coding exons were replaced by the lacZ reporter. To investigate further the myelin abnormalities in the absence of PMP22, sciatic nerves and dorsal root ganglion (DRG) neuron explant cultures from PMP22‐deficient mice were studied at various stages of myelination. Throughout the first 3 months of postnatal development, myelin protein and β4 integrin levels are dramatically reduced, whereas p75 and β1 integrin remain elevated. By immunostaining, the distributions of several glial proteins, including β4 integrin, the voltage‐gated potassium channel Kv1.1, and E‐cadherin, are altered. Schwann cells from PMP22‐deficient mice are able to produce limited amounts of myelin in DRG explant cultures, yet the internodal segments are dramatically fewer and shorter. The comparison of PMP22‐deficient mice with other PMP22 mutant models reveals that the decrease in β4 integrin is specific to an absence of PMP22. Furthermore, whereas lysosome‐associated membrane protein 1 and ubiquitin are notably up‐regulated in nerves of PMP22‐deficient mice, heat shock protein 70 levels remain constant or decrease compared with wild‐type or PMP22 mutant samples. Together these results support a role for PMP22 in the early events of peripheral nerve myelination. Additionally, although myelin abnormalities are a commonality among PMP22 neuropathic models, the underlying subcellular mechanisms are distinct and depend on the specific genetic abnormality. © 2006 Wiley‐Liss, Inc.
Growth and pathfinding of regenerating axons in the optic projection of adult fish
Authors: Becker Catherina G. , Becker Thomas
Published: November 2006

In contrast to mammals, teleost fish are able to regrow severed long‐range projection axons in the central nervous system (CNS), leading to recovery of function. The optic projection in teleost fish is used to study neuron‐intrinsic and environmental molecular factors that determine successful axon regrowth and navigation through a complex CNS pathway back to original targets. Here we review evidence for regeneration‐specific regulation and robust expression of growth‐ and pathfinding‐associated genes in regenerating retinal ganglion cell (RGC) axons of adult fish. The environment of the CNS in fish appears to contain few inhibitory molecules and at the same time a number of promoting molecules for axon regrowth. Finally, some environmental cues that are used as guidance cues for developing RGC axons are also present in continuously growing adult animals. These molecules may serve as guidance cues for the precise navigation of axons from newly generated RGCs in adult animals as well as of regenerating RGC axons after a lesion. The application of new molecular techniques especially to adult zebrafish, is likely to produce new insights into successful axonal regeneration in the CNS of fish and the absence thereof in mammals. © 2006 Wiley‐Liss, Inc.
Charged amino acid motifs flanking each extreme of the αM4 transmembrane domain are involved in assembly and cell‐surface targeting of the muscle nicotinic acetylcholine receptor
Authors: Roccamo A.M. , Barrantes F.J.
Published: November 2006

The αM4 transmembrane domain of the nicotinic acetylcholine receptor (AChR) is flanked by two basic amino acids (His408 and Arg429) located at its cytoplasmic‐ and extracellular‐facing extremes, respectively, at the level of the phospholipid polar head regions of the postsynaptic membrane. A series of single and double αM4 mutants (His408Ala, Arg429Ala, Arg429Glu, His408Ala/Arg429Ala, and His408Ala/Arg429Glu) of the adult muscle‐type AChR were produced and coexpressed with wild‐type β, δ, and ϵ subunits as stable clones in a mammalian heterologous expression system (CHO‐K1 cells). The mutants were studied by α‐bungarotoxin ([125I]α‐BTX) binding, fluorescence microscopy, and equilibrium sucrose gradient centrifugation. Cell‐surface [125I]α‐BTX binding diminished ∼40% in His408Ala and as much as 95% in the Arg429Ala mutant. Reversing the amino acid charge (e.g., Arg429Glu) abolished cell‐surface expression of AChR. Fluorescence microscopy disclosed that AChR was retained at the endoplasmic reticulum, with an enhanced occurrence of unassembled AChR species in the mutant clones. Centrifugation analysis confirmed the lack of fully assembled AChR pentamers in all mutants with the exception of His408Ala. We conclude that His408 and Arg429 in αM4 are involved in assembly and cell‐surface targeting of muscle AChR. Arg429 plays a more decisive role in these two processes, suggesting an asymmetric weight of the charged motifs at each extreme of the α subunit M4 transmembrane segment. © 2006 Wiley‐Liss, Inc.
Aβ(25–35) and its C‐ and/or N‐blocked derivatives: Copper driven structural features and neurotoxicity
Authors: Giuffrida Maria Laura , Grasso Giulia , Ruvo Menotti , Pedone Carlo , Saporito Angela , Marasco Daniela , Pignataro Bruno , Cascio Claudia , Copani Agata , Rizzarelli Enrico
Published: November 2006

The toxic properties of β‐amyloid protein, Aβ(1–42), the major component of senile plaques in Alzheimer's disease, depend on nucleation‐dependent oligomerization and aggregation. In addition, Aβ(1–42) toxicity is favored by the presence of trace metals, which affect the secondary structure of the peptide. A peptide comprising 11 residues within Aβ(1–42) [Aβ(25–35)] aggregates and retains the neurotoxic activity of Aβ(1–42). We have used both Aβ(25–35) and its C‐amidated or N‐acetylated/C‐amidated derivatives to investigate the role of copper(II) in modulating the conformation and aggregation state as well as the neurotoxic properties of amyloid peptides. Electrospray ionization mass spectrometry (ESI‐MS) and electron paramagnetic resonance (EPR) measurements were performed to verify the formation of copper(II)/Aβ(25–35) complexes and to determine the coordination mode, respectively. Aβ(25–35) and its derivatives were analyzed by circular dichroism spectroscopy to assess their secondary structure, subjected to thioflavine‐T (Th‐T) binding assay to reveal β‐sheet structured aggregates formation, and imaged by scanning force microscopy. Toxicity was assessed on mature cultures of rat cortical neurons. We found that β‐sheet‐structured species of Aβ(25–35) were neurotoxic, whereas the random‐coil‐structured derivatives were devoid of effect. Interestingly, copper promoted the random‐coil/β‐sheet transition of Aβ(25–35), with ensuing peptide toxicity, but it induced the toxicity of the N‐acetylated/C‐amidated derivative without affecting peptide folding. Moreover, copper did not influence either the folding or the activity of the C‐amidated Aβ(25–35), suggesting that blockade of the C‐terminus of Aβ peptides might be sufficient to prevent Aβ toxicity. © 2006 Wiley‐Liss, Inc.
Selective use of radioactive iodine in the postoperative management of patients with papillary and follicular thyroid carcinoma
Authors: Hay Ian D.
Published: November 2006

Radioiodine remnant ablation (RRA) was developed in the 1960s to “complete a thyroidectomy” in the initial management of papillary and follicular thyroid cancer. By the 1990s, it was claimed that RRA diminished recurrence rates in follicular cell‐derived cancer (FCDC) patients and decreased the cause‐specific mortality (CSM) in patients more than 40 years old at initial surgery. The international trend for the past decade has been towards routine RRA in most FCDC patients. Clinical guidelines have been produced by many societies, promoting such an aggressive stance. Since 1997, many papers have reported improved outcome in FCDC, when patients were subjected to RRA after bilateral lobar resection. However, during the same time‐period, it has been recognized that most FCDC patients are truly at “low‐risk” of developing life‐threatening recurrences. Accordingly, it has been suggested that rational therapy selection should lead to restricting aggressive therapy to those “high‐risk” FCDC patients, more predisposed to CSM. To date, no prospective controlled trials exist. Presently available outcome data is based on single institutional or multicenter retrospective studies. This article summarizes the available relevant reported data, and concludes that a selective use of RRA in the postoperative management of FCDC patients is rational, and should actually be encouraged. J. Surg. Oncol. 2006;94:692–700. © 2006 Wiley‐Liss, Inc.
Pathology and genetics of thyroid carcinoma
Authors: DeLellis Ronald A.
Published: November 2006

Carcinomas of the thyroid comprise a heterogeneous group of neoplasms with distinctive clinical and pathological characteristics. Over the past 15 years, the application of molecular technologies to the study of these neoplasms has elucidated critical genetic pathways associated with the development of specific thyroid tumor types. In papillary thyroid carcinoma (PTC), genetic events involve RET and TRK (rearrangements) and BRAF and RAS (mutations), although RAS mutations are uncommon except in the follicular variant of PTC. These genetic alterations, which rarely overlap in the same tumor, result in signaling abnormalities in the mitogen‐activated protein kinase pathway. In contrast, genetic alterations in follicular carcinomas include PAX8‐PPARγ translocations and RAS mutations while mutations of CTNNB1 and p53 have been implicated in the development and progression of poorly differentiated and undifferentiated (anaplastic) thyroid carcinomas. Germline mutations of RET are responsible for the development of heritable forms of medullary thyroid carcinoma (MTC) while somatic mutations of this oncogene are found in a significant proportion of sporadic MTCs. The results of these studies not only have provided additional approaches to thyroid tumor classification, but also have stimulated the development of novel approaches to tumor diagnosis and additional parameters for prognostic assessment and potential biologic therapeutic strategies. J. Surg. Oncol. 2006;94:662–669. © 2006 Wiley‐Liss, Inc.
Contemporary diagnostic approach to the thyroid nodule
Authors: Rosen Jennifer E. , Stone Michael D.
Published: November 2006

Thyroid nodules are common, with an estimated incidence of 5%–10% in the United States. The current gold standard for diagnosis is fine needle aspiration biopsy (FNAB). The incidence of indeterminate diagnoses varies from 10% to 25%. Surgical resection is usually indicated to exclude the diagnosis of cancer in these patients. However, only a minority (about 20%) of indeterminate thyroid nodules actually harbor a malignancy, resulting in surgery for diagnostic purposes alone in many patients. The increased detection of benign nodules and microcarcinomas reinforces the need for improved non‐operative methods to differentiate benign from malignant disease and discriminate low‐risk from high‐risk cancers. In this article we present a current, rational diagnostic approach to the patient with a thyroid nodule, evaluate new advances including thyroid genomic and predictor models, and propose the development of prospective trials to incorporate these new additions into clinical decision making. Given how many questions still exist for patients with thyroid nodules, partnership and collaboration, or the “bench to bedside” concept should find its way into most every thyroid surgeon and endocrinologist's lexicon. J. Surg. Oncol. 2006;94:649–661. © 2006 Wiley‐Liss, Inc.
Seminars: Controversies in the management of pediatric thyroid malignancy
Authors: Gingalewski Cynthia A. , Newman Kurt D.
Published: November 2006

Thyroid cancer in children is a rare malignancy with unusual biological behavior. It often presents at advanced stages, yet behaves in a benign manner, when compared to its adult counterpart. Surgeons have debated the risks and benefits of aggressive surgical procedures for over a decade. A consensus treatment plan for childhood thyroid cancer has not been achieved, however radical surgical procedures have, in general, fallen out of favor. The best operative and adjuvant strategy for these children can only be determined when a better understanding of the tumor cell biology and genetics of this cancer is known. This review discusses the current controversies in the management of differentiated childhood thyroid malignancy. J. Surg. Oncol. 2006;94:748–752. © 2006 Wiley‐Liss, Inc.