bluuebunny/crossref_metadata_2025 huggingface dataset:
Phosphatidic acid induces the differentiation of human acute promyelocytic leukemic cells into dendritic cell‐like
Authors: Jin Jun‐O , Park Hae‐Young , Kim Ja‐Woong , Park Joo‐In , Hong Young‐Seoub , Min Do Sik , Kwak Jong‐Young
Published: August 2006

We investigated whether phosphatidic acid (PA) can differentiate the promyelocytic leukemia (PML)‐retinoic acid receptor α (RARα)‐expressing acute promyelocytic leukemic cell line, NB4, to dendritic cell (DC)‐like cells. Dioctanoyl‐PA alone upregulated the expression of DC markers. The expression of DC markers on NB4 cells was potentiated by the overexpression of phospholipase D and upregulation was blocked by the addition of n‐butanol, an inhibitor of PA production. The expression of CD11c, CD83, and CCR7 in PA‐treated NB4 cells was further increased by tumor necrosis factor (TNF)‐α treatment. Increased functional capacities were also found in PA‐differentiated and TNF‐α‐activated NB4 cells with respect to changes in T‐cell proliferation, cytokine production, endocytic activity, and cytolytic capacity against undifferentiated NB4 cells. PA alone increased the phosphorylation of extracellular signal‐regulated kinase (ERK)‐1/2. The expression of DC markers was downregulated by PD98059, a specific inhibitor of ERK kinase or transient transfection of mutant‐ERK. The level of PML‐RARα fusion protein was decreased by PA treatment and PD98059 blocked the decrease of PML‐RARα. These results suggest that PA induces differentiation of NB4 cells into DC‐like cells and that the upregulation of antigen presenting cell markers is mediated by the activation of ERK and the downregulation of PML‐RARα levels. J. Cell. Biochem. 100: 191–203, 2007. © 2006 Wiley‐Liss, Inc.
Modeled microgravity stimulates osteoclastogenesis and bone resorption by increasing osteoblast RANKL/OPG ratio
Authors: Rucci Nadia , Rufo Anna , Alamanou Marina , Teti Anna
Published: August 2006

Mechanical unloading causes detrimental effects on the skeleton, but the underlying mechanisms are still unclear. We investigated the effect of microgravity on osteoblast ability to regulate osteoclastogenesis. Mouse osteoblast primary cultures were grown for 24 h at unit gravity or under simulated microgravity, using the NASA‐developed Rotating Wall Vessel bioreactor. Conditioned media (CM) from osteoblasts subjected to microgravity increased osteoclastogenesis and bone resorption in mouse bone marrow cultures. In these osteoblasts, the RANKL/OPG ratio was higher relative to 1g. Consistently, treatment with high concentrations of OPG‐inhibited osteoclastogenesis and bone resorption in the presence of CM arising from osteoblasts cultured under microgravity. Microgravity failed to affect osteoblast differentiation and function in the time frame of the experiment, as we found no effect on alkaline phosphatase mRNA and activity, nor on Runx2, osteocalcin, osteopontin, and collagen1A2 mRNA expression. In contrast, microgravity induced a time dependent increase of ERK‐1/2 phosphorylation, while phospho‐p38 and phospho‐JNK remained unchanged. Apoptosis, revealed by bis‐benzimide staining, was similar among the various gravity conditions, while it was increased under microgravity after treatment with the MEK‐1/2 inhibitor, PD98059, suggesting a protection role by ERK‐1/2 against cell death. In conclusion, microgravity is capable to indirectly stimulate osteoclast formation and activity by regulating osteoblast secretion of crucial regulatory factors such as RANKL and OPG. We hypothesize that this mechanism could contribute to bone loss in individuals subjected to weightlessness and other unloading conditions. J. Cell. Biochem. 100: 464–473, 2007. © 2006 Wiley‐Liss, Inc.
Hormonal induction of adipogenesis induces Skp2 expression through PI3K and MAPK pathways
Authors: Auld Corinth A. , Caccia Carla D. , Morrison Ron F.
Published: August 2006

We have previously shown that the F‐box protein, S‐phase kinase‐associated protein (Skp2) plays a mechanistic role in targeting the cell‐cycle inhibitor, p27 for degradation by the 26S proteasome during early stages of 3T3‐L1 adipocyte differentiation. Here, we demonstrate that protein levels of Skp2 and its accessory protein, Cks1 increased as density‐arrested preadipocytes re‐entered the cell cycle during clonal expansion, decreased with differentiation‐induced growth arrest, and became refractory to hormonal stimulation following the onset of terminal adipocyte differentiation. Component analysis revealed that while maximal Skp2/Cks1 protein accumulation required the complete differentiation cocktail, that insulin was principally involved. Skp2 mRNA accumulation was found to precede the increase in Skp2 protein and succeed the activation of Akt and Erk1/2, mediators of phosphatidylinositol‐3 kinase (PI3K) and mitogen‐activated protein kinase (MAPK) signal transduction pathways, respectively. Using specific inhibitors, we found that while activation of both pathways was required for maximal expression, PI3K signaling was primarily responsible for the increase in Skp2/Cks1 accumulation. The increase in Skp2 mRNA was notable 4 h following hormonal stimulation, plateaued by 12 h during mid‐G1 phase progression, and occurred without change to mRNA stability. We further demonstrate that luciferase activity, originating from a pGL3 vector containing 2.4 kb of the Skp2 promoter, increased 2.5‐fold with hormonal stimulation. This increase in promoter activity was markedly suppressed following PI3K and MAPK blockade. Deletion studies indicate that responsive elements were located within the proximal Skp2 promoter. These data demonstrate that Skp2 is transcriptionally regulated by PI3K and MAPK pathways as 3T3‐L1 preadipocytes transition from quiescence to proliferation during adipocyte hyperplasia. J. Cell. Biochem. 100: 204–216, 2007. © 2006 Wiley‐Liss, Inc.
Regulated expression of syndecan‐4 in rat calvaria osteoblasts induced by fibroblast growth factor‐2
Authors: Song Shu Jun , Cool Simon M. , Nurcombe Victor
Published: August 2006

Fibroblast growth factor‐2 (FGF2) is a member of a prominent growth factor family that drives proliferation in a wide variety of cell types, including osteoblasts. The binding and signal transduction triggered by these mitogens is dependent on glycosaminoglycan (GAG) sugars, particularly of the heparan sulfate (HS) class. These are secreted in proteoglycan (PG) complexes, some of which become FGF co‐receptors. The syndecans, the transmembrane forms of HSPG of which there are four members, act as multifunctional receptors for a variety of ligands involved in cell‐extracellular matrix (ECM) adhesion as well as growth factor binding. To understand the role of syndecans in developing osteoblasts, the effects of exogenous FGF2 on syndecan expression were examined using primary rat calvarial osteoblasts. All four syndecan mRNAs were expressed in the osteoblasts, although only syndecan‐4 was upregulated by FGF2 treatment in a dose‐dependent manner. This upregulation could be abrogated by pretreatment with the protein synthesis inhibitor cycloheximide, suggesting that the upregulation of syndecan‐4 by FGF2 is not a primary response. Osteoblast proliferation and mineralization were enhanced by exogenous FGF2 treatment, but could be specifically diminished by anti‐syndecan‐4 antibody pretreatment. This treatment also blocked FGF2‐induced extracellular signal‐regulated kinase activation, but not the expression of the bone‐specific transcription factor Runx2. These results demonstrate that mitogen‐triggered syndecan‐4 expression is an intrinsic part of the pathways subtending osteoblast proliferation and mineralization. J. Cell. Biochem. 100: 402–411, 2007. © 2006 Wiley‐Liss, Inc.
Increase in β1‐6 GlcNAc branching caused by N‐acetylglucosaminyltransferase V directs integrin β1 stability in human hepatocellular carcinoma cell line SMMC‐7721
Authors: Wang Liying , Liang Yulong , Li Zengxia , Cai Xiumei , Zhang Wen , Wu Guoqiang , Jin Jiawei , Fang Zhengyu , Yang Yong , Zha Xiliang
Published: August 2006

In this study, an enzymatic inactive mutant of GnT‐V (ΔcGnT‐V) was constructed and transfected in SMMC 7721 cell line. Integrin β1 in ΔcGnT‐V transfectants (Δc‐7721) showed attenuation of the number of β1‐6 GlcNAc branching, whereas those in wtGnT‐V transfectants (wt‐7721) presented a β1‐6 GlcNAc‐rich pattern. High integrin β1 expression was observed in wt‐7721 compared with mock cells (7721 cell transfected with the vector pcDNA3), while transfection of ΔcGnT‐V decreased the integrin β1 expression, despite of no significant changes on integrin β1 mRNA level in these cell lines. Pulse‐chase experiment showed that Integrin β1 in Δc‐7721 was prone to quick degradation and its half‐life was less than 3 h, on the contrary, the alleviating degradation of β1 subunit was observed in wt‐7721 where the β1 subunit half‐life was about 16 h, meanwhile, the degradation rate of β1 subunit in mock cells was in between, about 10 h. More effective in promoting cell migration toward fibronectin and invasion through Matrigel was observed in wt‐7721 while this was almost suppressed in Δc‐7721. Our results suggest that the addition of β1‐6 GlcNAc branching caused more fully glycosylated mature form on integrin β1 and inhibited β1 protein degradation. Glycosylation caused by GnT‐V directs integrin β1 stability and more delivery to plasma membrane, subsequently promotes Fn‐based cell migration and invasion. J. Cell. Biochem. 100: 230–241, 2007. © 2006 Wiley‐Liss, Inc.
Block designs which are divisible in multiple ways
Authors: Mendelsohn N. S.
Published: August 2006

Constructions are given for the production of symmetric block designs whose point set is divisible (partitionable) in several ways. Some properties of the dual design are also considered. © 2006 Wiley Periodicals, Inc. J Combin Designs 14: 415–422, 2006
Acrosome‐specific gene AEP1: Identification, characterization and roles in spermatogenesis
Authors: Luk John M. , Lee Nikki P.Y. , Shum Cathy K. , Lam Brian Y. , Siu Annie F.M. , Che Chi‐Ming , Tam Po‐Chor , Cheung Annie N.Y. , Yang Z.M. , Lin Yi‐Nan , Matzuk Martin M. , Lee Kai‐Fai , Yeung William S.B.
Published: August 2006

Spermatogenesis is a tightly regulated process leading to the development of spermatozoa. To elucidate the molecular spermatogenic mechanisms, we identified an acrosome‐specific gene AEP1 in spermatids, which is located in rat chromosome 17p14 with a transcript size of 3,091 bp encoding a signal peptide, zinc finger‐like motif, coiled‐coil region, several predicted glycosylation and phosphorylation sites. Northern blot and RT‐PCR analyses revealed the restricted expression of AEP1 to the testis only. In postnatal rat testes, AEP1 mRNA became detectable from postnatal 25 dpp (round spermatids) and onwards. By using in situ hybridization (ISH) and flow cytometry‐fluorescent ISH, only the haploid spermatids yielded the positive AEP1 signal. Immunohistochemistry showed that AEP1 was expressed in the acrosomal cap of late‐staged germ cells in rat testis, and co‐localized with the acrosomal marker, peanut agglutinin. The spatial expression of AEP1 immunoreactivity in testis was conserved among diverse mammalian species (rat, pig, monkey, human). To further study its roles in spermatogenesis, we showed AEP1 and β‐actin was associated together in complex by co‐immunoprecipitation in adult germ cells and by immunofluorescence assay in isolated spermatozoon. In human testes diagnosed with hypospermatogenesis, lower expression of AEP1 was observed, whereas there was no detectable signal in undescended testes. In short, AEP1 is an evolutionary‐conserved acrosome‐specific gene and likely functions in acrosome‐cap formation. J. Cell. Physiol. 209: 755–766, 2006. © 2006 Wiley‐Liss, Inc.
Bridging behavior and physiology: Ion‐channel perspective on mushroom body‐dependent olfactory learning and memory in Drosophila
Authors: Gasque Gabriel , Labarca Pedro , Delgado Ricardo , Darszon Alberto
Published: August 2006

An important body of evidence documents the differential expression of ion channels in brains, suggesting they are essential to endow particular brain structures with specific physiological properties. Because of their role in correlating inputs and outputs in neurons, modulation of voltage‐dependent ion channels (VDICs) can profoundly change neuronal network dynamics and performance, and may represent a fundamental mechanism for behavioral plasticity, one that has received less attention in learning and memory studies. Revisiting three paradigmatic mutations altering olfactory learning and memory in Drosophila (dunce, leonardo, amnesiac) a link was established between each mutation and the operation of VDICs in Kenyon cells, the intrinsic neurons of the mushroom bodies (MBs). In Drosophila, MBs are essential to the emergence of olfactory associative learning and retention. Abnormal ion channel operation might underlie failures in neuronal physiology, and be crucial to understand the abnormal associative learning and retention phenotypes the mutants display. We also discuss the only case in which a mutation in an ion channel gene (shaker) has been directly linked to olfactory learning deficits. We analyze such evidence in light of recent discoveries indicating an unusual ion current profile in shaker mutant MB intrinsic neurons. We anticipate that further studies of acquisition and retention mutants will further confirm a link between such mutations and malfunction of specific ion channel mechanisms in brain structures implicated in learning and memory. J. Cell. Physiol. 209: 1046–1053, 2006. © 2006 Wiley‐Liss, Inc.
Purinergic pathway suppresses the release of .NO and stimulates proteoglycan synthesis in chondrocyte/agarose constructs subjected to dynamic compression
Authors: Chowdhury T.T. , Knight M.M.
Published: August 2006

Mechanical loading plays a fundamental role in the physiological and pathological processes of articular cartilage. The application of dynamic compression to chondrocytes cultured in agarose, downregulates the release of nitric oxide (NO) and enhances cell proliferation and proteoglycan synthesis. We hypothesize that the observed metabolic changes in response to dynamic compression involve a purinergic signaling pathway. Chondrocyte/agarose constructs were subjected to dynamic compression (15%, 1 Hz, 48 h) in the presence of antagonists for the purinergic pathway. Gadolinium was used as a putative inhibitor of stretch‐activated calcium ion channels including adenosine 5′‐triphophate (ATP) release channels; suramin was employed as a P2 receptor antagonist and apyrase was used to catalyze the hydrolysis of extracellular ATP. The data presented demonstrate that in the absence of the inhibitor, dynamic compression suppressed .NO release. Treatment with gadolinium and suramin caused a compression‐induced upregulation of .NO release, a response abolished with apyrase. Compression‐induced stimulation of cell proliferation was reversed with gadolinium, suramin, or apyrase. By contrast, compression‐induced stimulation of proteoglycan synthesis was abolished under all treatment conditions. Thus, the purinergic pathway is important in suppressing the release of .NO and stimulation of proteoglycan synthesis. Indeed, high levels of .NO could trigger a downstream catabolic response and mediate the compression‐induced inhibition of cell proliferation. The current study demonstrates for the first time the importance of a purinergic pathway in mediating the metabolic response to dynamic compression and suppressing an inflammatory effect. J. Cell. Physiol. 209: 845–853, 2006. © 2006 Wiley‐Liss, Inc.
Insulin restores glucose inhibition of adenosine transport by increasing the expression and activity of the equilibrative nucleoside transporter 2 in human umbilical vein endothelium
Authors: Muñoz Gonzalo , Martín Rody San , Farías Marcelo , Cea Luis , Vecchiola Andrea , Casanello Paola , Sobrevia Luis
Published: August 2006

L‐Arginine transport and nitric oxide (NO) synthesis (L‐arginine/NO pathway) are stimulated by insulin, adenosine or elevated extracellular D‐glucose in human umbilical vein endothelial cells (HUVEC). Adenosine uptake via the human equilibrative nucleoside transporters 1 (hENT1) and 2 (hENT2) has been proposed as a mechanism regulating adenosine plasma concentration, and therefore its vascular effects in human umbilical veins. Thus, altered expression and/or activity of hENT1 or hENT2 could lead to abnormal physiological plasma adenosine level. We have characterized insulin effect on adenosine transport in HUVEC cultured in normal (5 mM) or high (25 mM) D‐glucose. Insulin (1 nM) increased overall adenosine transport associated with higher hENT2‐, but lower hENT1‐mediated transport in normal D‐glucose. Insulin increased hENT2 protein abundance in normal or high D‐glucose, but reduced hENT1 protein abundance in normal D‐glucose. Insulin did not alter the reduced hENT1 protein abundance, but blocked the reduced hENT1 and hENT2 mRNA expression induced by high D‐glucose. Insulin effect on hENT1 mRNA expression in normal D‐glucose was blocked by NG‐nitro‐L‐arginine methyl ester (L‐NAME, NO synthase inhibitor) and mimicked by S‐nitroso‐N‐acetyl‐L,D‐penicillamine (SNAP, NO donor). L‐NAME did not block insulin effect on hENT2 expression. In conclusion, insulin stimulation of overall adenosine transport results from increased hENT2 expression and activity via a NO‐independent mechanism. These findings could be important in hyperglycemia‐associated pathological pregnancies, such as gestational diabetes, where plasma adenosine removal by the endothelium is reduced, a condition that could alter the blood flow from the placenta to the fetus affecting fetus growth and development. J. Cell. Physiol. 209: 826–835, 2006. © 2006 Wiley‐Liss, Inc.
Diphosphorylation of the myosin regulatory light chain enhances the tension acting on stress fibers in fibroblasts
Authors: Mizutani Takeomi , Haga Hisashi , Koyama Yoshikazu , Takahashi Masayuki , Kawabata Kazushige
Published: August 2006

Regulation of the contractile force is crucial for cell migration, cell proliferation, and maintenance of cell morphology. Phosphorylation of the myosin II regulatory light chain (MRLC) is involved in these processes. To show whether the diphosphorylation of MRLC increases the tension acting on stress fibers, changes in the stiffness of fibroblasts expressing wild‐type MRLC and a mutant type, which cannot be diphosphorylated, on treatment with lysophosphatidic acid (LPA) were examined by a mechanical‐scanning probe microscope (M‐SPM). The LPA treatment increased cellular stiffness in the wild‐type MRLC expressing cells, while it had no effect on the mutated cells. Immunostaining showed that LPA stimulation induced the diphosphorylation of MRLC. These results suggest that the diphosphorylation of MRLC enhances the tension acting on stress fibers. J. Cell. Physiol. 209: 726–731, 2006. © 2006 Wiley‐Liss, Inc.
Ultrastructure of cyst shell and underlying membranes of three strains of the brine shrimp Artemia (Branchiopoda: Anostraca) from South India
Authors: Sugumar V. , Munuswamy N.
Published: August 2006

The cyst of Artemia has shell and membranous coverings over the embryo. The membranous coverings have special adaptive features to allow the physical changes accompanying repeated hydration and dehydration cycles that might occur and adversely influence postembryonic development. Whole and slices of cryptobiotic cysts were processed for electron microscopy to study the internal details and to compare the morphological architecture of three Artemia strains of South India. Surface topography of scanning electron microscopic (SEM) studies revealed distinct button shaped structures on the cyst of Puthalam strain. Transmission electron microscopic (TEM) studies of the cysts displayed the conventional pattern of anostracan crustaceans with outer cortex and alveolar layer, cuticular membranes, and the cytoplasmic inclusions namely nucleus, yolk droplets, lipoid bodies, and mitochondria. The prominent wavy outer cortex layer of Puthalam cysts corroborates the results of SEM studies. Microsc. Res. Tech. © 2006 Wiley‐Liss, Inc.
Characterisation of a P140K mutant O6‐methylguanine‐DNA‐methyltransferase (MGMT)‐expressing transgenic mouse line with drug‐selectable bone marrow
Authors: Kramer Belinda A. , Lemckert Frances A. , Alexander Ian E. , Gunning Peter W. , McCowage Geoffrey B.
Published: August 2006

Background

Gene transfer of the P140K mutant of O6‐methylguanine‐DNA‐methyltransferase (MGMT(P140K)) into hematopoietic stem cells (HSC) provides a mechanism for drug resistance and the selective expansion of gene‐modified cells in vivo. Possible clinical applications for this strategy include chemoprotection to allow dose escalation of alkylating chemotherapy, or combining MGMT(P140K) expression with a therapeutic gene in the treatment of genetic diseases. Our aim is to use MGMT(P140K)‐driven in vivo selection to develop allogeneic micro‐transplantation protocols that rely on post‐engraftment selection to overcome the requirement for highly toxic pre‐transplant conditioning, and to establish and maintain predictable levels of donor/recipient chimerism.

Methods

Using stably transfected murine embryonic stem (ES) cells, we have generated a C57BL/6 transgenic mouse line with expression of MGMT(P140K) within the hematopoietic compartment for use as a standard source of donor HSC in such models. Functional characterisation of transgene expression was carried out in chemotherapy‐treated transgenic mice and in allogeneic recipients of transgenic HSC.

Results

Expression of the transgene provided chemoprotection and allowed in vivo selection of MGMT(P140K)‐expressing cells in transgenic mice after exposure to O6‐benzylguanine (BG) and N,N′‐bis(2‐chloroethyl)‐N‐nitrosourea (BCNU). In an allogeneic transplant experiment in which transgenic HSC were engrafted into 129 strain recipients following low intensity conditioning (Busulfan, anti‐CD8, anti‐CD40Ligand), MGMT(P140K)‐expressing cells could be selected using chemotherapy.

Conclusions

This MGMT(P140K) transgenic mouse line provides a useful source of drug‐selectable donor cells for the development of non‐myeloablative allogeneic transplant models in which variation in transplant conditioning elements can be investigated independently of gene transfer efficiency. Copyright © 2006 John Wiley & Sons, Ltd.
High specificity of V3 serotyping among human immunodeficiency virus type‐1 subtype C infected patients with varying disease status and viral phenotype
Authors: Walker Polly R. , Cilliers Tonie , Choge Isaac A. , Taylor Natasha , Cohen Sarah S. , Morris Lynn
Published: August 2006

V3 serotyping is a technique for determining HIV‐1 genetic subtype based on the binding of antibodies from patient sera or plasma to synthetic V3 peptides derived from subtype consensus sequences. Variation in the performance of this assay has been attributed to V3 sequence heterogeneity, the degree of which varies with patient disease progression, virus co‐receptor usage, and genetic subtype. This study assessed the performance of a competitive peptide enzyme immunoassay (cPEIA) in samples from HIV‐1 subtype C infected patients with varying disease profiles, including those with syncytium (SI) and non‐syncytium‐inducing (NSI) viruses. Out of 90 sera tested, 94.4% reacted strongly against the subtype C peptide. There was no significant difference in assay sensitivity among samples from advanced AIDS patients in which humoral immune response may be lower, nor among SI viruses which carry changes in the V3 sequence. Four samples were found to be cross‐reactive with other subtypes and one acutely infected patient sample was non‐reactive due to low anti‐gp120 antibody titers. A significantly higher number of samples showed secondary reactivity to subtype A, compared to other subtypes (P < 0.005). In conclusion, the assay was able to identify HIV‐1 subtype C infection with a high level of sensitivity (94%) irrespective of the stage of disease and therefore provides a valuable resource for the large‐scale epidemiological monitoring of the spread of HIV‐1 subtypes in South Africa. J. Med. Virol. 78:1262–1268, 2006. © 2006 Wiley‐Liss, Inc.
Low HBV‐DNA levels in end‐stage renal disease patients with HBeAg‐negative chronic hepatitis B
Authors: Moutinho Renata S. , Perez Renata M. , Medina‐Pestana José O. , Figueiredo Mauro S. , Koide Sandra , Alberto Fernando Lopes , Silva Antonio Eduardo B. , Ferraz Maria Lucia G.
Published: August 2006

In end‐stage renal disease patients treated by hemodialysis with HBeAg‐negative chronic hepatitis B virus (HBV) infection, the evaluation of the presence of viral replication is essential in the assessment for renal transplantation. Data on HBV viral load, prevalence of precore mutations, as well as the influence of HCV coinfection on HBV‐DNA levels in this group of patients is scarce. The aim of this study was to determine the HBV viral load in HBsAg‐positive/HBeAg‐negative hemodialysis patients; to compare HBV‐DNA levels between isolated HBV infection carriers and HBV‐HCV coinfected patients, and to evaluate the prevalence of precore mutations in these patients. Fifty hemodialysis patients with chronic HBeAg‐negative HBV infection were studied. Viral load was determined by PCR (Amplicor HBV Monitor‐Roche). The detection of precore mutations was made by sequencing. Of a total of 50 patients, 76% were male, with a mean age of 44 ± 11 years. Anti‐HCV was positive in 56% of patients. HBV‐DNA was undetectable in 58% of patients; 24% had HBV‐DNA <10,000 copies/ml, 12% between 10,000–100,000 copies/ml, and only 6% had HBV‐DNA >100,000 copies/ml. There was no difference in the viral load of patients infected only by HBV and HBV‐HCV co‐infected patients (P = 0.96). Precore mutations were detected in only 8% of cases. In conclusion, hemodialysis patients with HBeAg‐negative HBV infection had a low viral load. Precore mutations were infrequent and the presence of anti‐HCV has not influenced the levels of HBV‐DNA. J. Med. Virol. 78:1284–1288, 2006. © 2006 Wiley‐Liss, Inc.
Microarray assay for detection and discrimination of Orthopoxvirus species
Authors: Ryabinin Vladimir A. , Shundrin Leonid A. , Kostina Elena B. , Laassri Majid , Chizhikov Vladimir , Shchelkunov Sergei N. , Chumakov Konstantin , Sinyakov Alexander N.
Published: August 2006

A microarray method was developed for simultaneous detection and identification of six species of Orthopoxvirus (OPV) including Variola, Monkeypox, Cowpox, Camelpox, Vaccinia, and Ectromelia viruses. The method allowed us to discriminate OPV species from varicella‐zoster virus (VZV), Herpes Simplex 1 virus (HSV‐1), and Herpes Simplex 2 virus (HSV‐2) that cause infections with clinical manifestations similar to OPV infections. The nucleotide sequences of the C23L/B29R and the B19R genes identified for 86 and 72 different OPV strains, respectively, were used to design species‐specific microarray oligonucleotide probes (oligoprobes). The microarray also contained several oligoprobes selected from the ORF31, US4, and US5 genes of VZV, HSV‐1, and HSV‐2, respectively. The samples (from HSVs or OPVs) of ssDNAs for analyses were prepared by using asymmetric PCR followed by chemical labeling of ssDNA with Cy3 dye. DNA from 52 samples of various OPV species, two isolates of VZV, two of HSV‐1, and three of HSV‐2 were tested using the developed microarray assay; all tested viruses were accurately identified. To ensure the robustness of the microarray assay, three additional unrelated variola virus strains with unknown sequences of the C23L/B29R and the B19R genes were tested. In each instance the microarray unambiguously identified them as Variola virus species. The results obtained in this study demonstrated that this new microarray method is a valuable tool for the rapid and accurate detection and differentiation of these important viral pathogens. J. Med. Virol. 78:1325–1340, 2006. © 2006 Wiley‐Liss, Inc.
Response to long‐term lamivudine treatment in patients infected with hepatitis B virus genotypes A, B, and C
Authors: Kobayashi Mariko , Suzuki Fumitaka , Akuta Norio , Suzuki Yoshiyuki , Arase Yasuji , Ikeda Kenji , Hosaka Tetsuya , Sezaki Hitomi , Kobayashi Masahiro , Iwasaki Satomi , Sato Junko , Watahiki Sachiyo , Miyakawa Yuzo , Kumada Hiromitsu
Published: August 2006

Response to lamivudine treatment longer than 1 year was compared in 15 patients persistently infected with hepatitis B virus (HBV) genotype A, 38 with genotype B, and 449 with genotype C. Patients with genotype A were younger (median age 37 [range 24–49] vs. 47 [24–67] or 44 [18–73], P = 0.015), possessed hepatitis B e antigen (HBeAg) more frequently (73% vs. 21% or 56%, P < 0.001) and HBV DNA in higher levels (8.6 [6.1–8.7] vs. 6.5 [<3.7–8.7] or 6.5 [<3.7–8.7] log genome equivalents (LGE)/ml, P = 0.024) than those with genotype B or C. During lamivudine, YMDD mutants (89% vs. 53% or 42%, P = 0.0001) and breakthrough hepatitis developed more often (47% vs. 21% or 29%, P = 0.023) in patients with genotype A than B or C. YMDD mutants elicited more frequently in patients with genotype A than B or C who were positive (82% [9/11] vs. 25% [2/8] or 48% [117/245], P = 0.037) or negative for HBeAg (75% [3/4] vs. 30% [9/30] or 33% [68/204], P = 0.003). HBeAg (hazard ratio 2.1 [95% confidence interval 1.53–2.92], P < 0.001) and genotype A (2.78 [1.08–7.12], P = 0.034) enhanced the emergence of YMDD mutants by the Cox proportional hazard model. The risk for breakthrough hepatitis was increased by the baseline alanine aminotransferase level <500 IU/L (2.56 [1.82–5.50], P = 0.018), HBeAg (2.11 [1.40–3.16], P < 0.001), cirrhosis (1.92 [1.24–2.97], P = 0.004) and HBV DNA ≥8.0 LGE/ml (1.57 [1.04–2.36], P = 0.03); it was influenced by genotypes only in patients with HBeAg. In conclusion, HBV genotypes help in predicting response to long‐term lamivudine treatment and development of YMDD mutants in patients with chronic hepatitis B. J. Med. Virol. 78:1276–1283, 2006. © 2006 Wiley‐Liss, Inc.
Distinct patterns of evolution between respiratory syncytial virus subgroups A and B From New Zealand isolates collected over thirty‐seven years
Authors: Matheson James W. , Rich Fenella J. , Cohet Catherine , Grimwood Keith , Huang Q. Sue , Penny David , Hendy Michael D. , Kirman Joanna R.
Published: August 2006

Respiratory syncytial virus (RSV) is the most important cause of viral lower respiratory tract infections in infants and children worldwide. In New Zealand, infants with RSV disease are hospitalized at a higher rate than other industrialized countries, without a proportionate increase in known risk factors. The molecular epidemiology of RSV in New Zealand has never been described. Therefore, we analyzed viral attachment glycoprotein (G) gene sequences from 106 RSV subgroup A isolates collected in New Zealand between 1967 and 2003, and 38 subgroup B viruses collected between 1984 and 2004. Subgroup A and B sequences were aligned separately, and compared to sequences of viruses isolated from other countries during a similar period. Genotyping and clustering analyses showed RSV in New Zealand is similar and temporally related to viruses found in other countries. By quantifying temporal clustering, we found subgroup B viruses clustered more strongly than subgroup A viruses. RSV B sequences displayed more variability in stop codon usage and predicted protein length, and had a higher degree of predicted O‐glycosylation site changes than RSV A. The mutation rate calculated for the RSV B G gene was significantly higher than for RSV A. Together, these data reveal that RSV subgroups exhibit different patterns of evolution, with subgroup B viruses evolving faster than A. J. Med. Virol. 78:1354–1364, 2006. © 2006 Wiley‐Liss, Inc.
Modulation of epitope‐specific anti‐hepatitis C virus E2 (anti‐HCV/E2) antibodies by anti‐viral treatment
Authors: Mancini Nicasio , Carletti Silvia , Perotti Mario , Romanò Luisa , Di Stefano Craxì Rosellina , Craxì Antonio , Zanetti Alessandro R. , Clementi Massimo , Burioni Roberto
Published: August 2006

The dynamic features of three specific anti‐hepatitis C virus (HCV) antibody subpopulations directed against different conformational epitopes of the viral E2 protein (HCV/E2) have been evaluated in patients with primary and persistent HCV infection; the three subpopulations are present in patients infected with different HCV genotypes and have shown a different activity using a pseudovirus neutralization assay (antibodies e301 and e137 exhibiting high neutralizing activity, while antibody e509 enhancement of HCV infectivity). In sequential samples from five patients with primary HCV infection and different virological outcome, all samples tested negative with the single exception of the e509 antibody in a patient not clearing the virus. In sequential samples from 28 patients with persistent infection under treatment with pegylated interferon‐α plus ribavirin (14 sustained virological responders and 14 non‐responders), the therapy did not selectively influence titers of the two neutralizing antibody subpopulations; otherwise, a net increase of the e509 antibody subpopulation related to enhancement of HCV infectivity was observed in non‐responders, but not in sustained virological responders (P = 0.0156). This increase was not related to the trend of total anti‐HCV/E2 response. The data indicate that a specific antibody response against these epitopes is elicited only late during the infection, thus not influencing virus clearance during primary infection, and that a selective increase of the antibody subpopulation enhancing virus infectivity is observed only in the cohort of patients not responding to antiviral therapy. J. Med. Virol. 78:1304–1311, 2006. © 2006 Wiley‐Liss, Inc.
Glacial modification of granite tors in the Cairngorms, Scotland
Authors: Hall A. M. , Phillips W. M.
Published: August 2006

A range of evidence indicates that many granite tors in the Cairngorms have been modified by the flow of glacier ice during the Pleistocene. Comparisons with SW England and the use of a space–time transformation across 38 tor groups in the Cairngorms allow a model to be developed for progressive glacial modification. Tors with deeply etched surfaces and no, or limited, block removal imply an absence of significant glacial modification. The removal of superstructure and blocks, locally forming boulder trains, and the progressive reduction of tors to stumps and basal slabs represent the more advanced stages of modification. Recognition of some slabs as tor stumps from which glacial erosion has removed all superstructure allows the original distribution of tors to be reconstructed for large areas of the Cairngorms. Unmodified tors require covers of non‐erosive, cold‐based ice during all of the cold stages of the Middle and Late Pleistocene. Deformation beneath cold‐based glacier ice is capable of the removal of blocks but advanced glacial modification requires former wet‐based glacier ice. The depth of glacial erosion at former tor sites remains limited largely to the partial or total elimination of the upstanding tor form. Cosmogenic nuclide exposure ages (Phillips et al., 2006) together with data from weathering pit depths (Hall and Phillips, 2006), from the surfaces of tors and large erratic blocks require that the glacial entrainment of blocks from tors occurred in Marine Isotope Stages (MIS) 4–2, 6 and, probably, at least one earlier phase. The occurrence of glacially modified tors on or close to, the main summits of the Cairngorms requires full ice cover over the mountains during these Stages. Evidence from the Cairngorms indicates that tor morphology can be regarded as an important indicator of former ice cover in many formerly glaciated areas, particularly where other evidence of ice cover is sparse. Recognition of the glacial modification of tors is important for debates about the former existence of nunataks and refugia. Copyright © 2006 John Wiley & Sons, Ltd.
RNA Interference as a tool to study the function of MAD2 in mouse oocyte meiotic maturation
Authors: Wang Jian‐Ying , Lei Zi‐Li , Nan Chang‐Long , Yin Shen , Liu Jinghe , Hou Yi , Li Yun‐Long , Chen Da‐Yuan , Sun Qing‐Yuan
Published: August 2006

Spindle checkpoint proteins control entry into anaphase and chromosome segregation. As a member of spindle checkpoint proteins, MAD2 takes a central role in the regulation of anaphase onset and genome integrity. Here, we used MAD2 siRNA transfection approach to study MAD2 functions during mouse oocyte meiotic maturation in vitro. Real‐time PCR and laser scanning confocal microscopy showed that we successfully downregulated MAD2 transcript and protein expression. We further demonstrated that MAD2 downregulation resulted in a shortened duration of meiosis I and meiotic spindle abnormality, suggesting the function of MAD2 in mouse oocyte meiotic maturation. We also showed that MAD2 interference to some extent decreased GVBD rate, but increased apoptosis in mouse oocytes. In conclusion, our study shows that siRNA transfection is an effective tool to study MAD2 functions, and our results provide further evidence for the role of MAD2 as a spindle checkpoint protein in mouse oocytes. Mol. Reprod. Dev. 74: 116–124, 2007. © 2006 Wiley‐Liss, Inc.
Evidence for blood chimerism in dizygotic spontaneous twin pregnancy discordant for Down syndrome
Authors: Shalev S. A. , Shalev E. , Pras E. , Shneor Y. , Gazit E. , Yaron Y. , Loewenthal R.
Published: August 2006

Background

A monochorionic‐diamniotic placenta (MCDAP) is rare in dizygotic (DZ) twinning. All reported cases have been documented in pregnancies achieved by the induction of ovulation alone or during the IVF cycle.

Methods and Results

We report a spontaneous pregnancy in a 39‐year‐old patient with evidence of MCDAP in DZ twins, discordant for trisomy 21. The first and second‐trimester sonographic scans indicated male twins with MCDAP. Amniocentesis, performed because of advanced maternal age, revealed a normal karyotype in one fetus, and trisomy 21 in the other. Molecular studies, performed in order to confirm the zygosity and chorionicity, demonstrated that the fetuses were DZ. In order to identity the affected twin, a detailed sonographic examination was repeated, but no abnormal findings associated with Down syndrome were demonstrated in any of the fetuses. Therefore, umbilical cord blood samples were obtained from both fetuses. Chromosomal analysis revealed in both fetuses two cell lines: a normal cell line of 46,XY and a 47,XY,+ 21 cell line, in 65 and 80% of the cells, respectively. This result was independently confirmed by both FISH and G‐banding. DNA extracted from both cord blood samples demonstrated an admixture of two distinct genotypes in each sample.

Conclusions

We propose that this case represents a monochorionic‐dizygotic twin pregnancy with blood chimerism. The most plausible mechanism underlying this phenomenon is placental fusion early in pregnancy, resulting in an architecturally single placenta originating from two distinct zygotes. The newly formed blood vessels created anastomoses between the DZ twins and allowed reciprocal blood chimerism between the normal and the trisomic twin. Copyright © 2006 John Wiley & Sons, Ltd.
The influence of faith and religion and the role of religious and community leaders in prenatal decisions for sickle cell disorders and thalassaemia major
Authors: Ahmed Shenaz , Atkin Karl , Hewison Jenny , Green Josephine
Published: August 2006

Objectives

Religion is believed to have a significant impact on individuals from minority ethnic groups when making decisions about prenatal genetic screening, prenatal diagnosis and termination of pregnancy. This study aimed to explore the views of individuals from South‐Asian and African‐Caribbean communities towards termination of pregnancy for sickle cell disorders and thalassaemia major and the influence of (1) faith and religion, (2) perceived severity of the conditions, and (3) religious and community leaders.

Methods

The study explored the views of (1) individuals from four faith communities (Pakistani Muslims, Indian Hindus, Indian Sikhs, African‐Caribbean Christians), using eight focus groups, and (2) parents of children with sickle cell disorders and thalassaemia major, using two focus groups and three interviews.

Results

Participants' accounts suggest that they generally considered religion and faith as an important factor in the decision‐making process, but the perceived severity of the condition would play a more important role. Religious and community leaders were believed to have little role to play in the decision‐making process.

Conclusion

The findings emphasise the importance of recognising diversity within different faith groups and moving away from stereotypical views based on people's ethnicity or religion, and to consider the beliefs and preferences of individuals. Copyright © 2006 John Wiley & Sons, Ltd.
German lexical personality factors: relations with the HEXACO model
Authors: Ashton Michael C. , Lee Kibeom , Marcus Bernd , De Vries Reinout E.
Published: August 2006

We correlated the scales of the HEXACO Personality Inventory (HEXACO‐PI) with adjective scale markers of factors previously obtained in indigenous lexical studies of personality structure in the German language. Self‐ratings obtained from a sample of 323 German participants showed a pattern of strong convergent and weak discriminant correlations, supporting the content‐based interpretation of the German lexical factors in terms of the HEXACO dimensions. Notably, convergent correlations were strong for both the broader and the narrower variants of the Honesty‐Humility factor as observed in German lexical studies. Also, convergent correlations for HEXACO Openness to Experience were, as expected, stronger for German adjectives describing a creative and intellectual orientation than for German adjectives describing intellectual ability. Copyright © 2006 John Wiley & Sons, Ltd.
Synthesis and characterization of a helical step‐ladder polyarylene
Authors: Nehls Benjamin S. , Galbrecht Frank , Brauer David J. , Lehmann Christian W. , Scherf Ullrich , Farrell Tony
Published: August 2006

A helical step‐ladder polyarylene incorporating chiral (R)‐2,2′‐dioctoxy‐1,1′‐binaphthyl units was synthesized for the first time. The first step involved the preparation of a precursor poly(arylene ketone) via a palladium‐mediated Suzuki‐type cross‐coupling reaction with the aid of microwave heating. Two polymer‐analog reaction steps, the reduction of the keto groups to tertiary alcohol functionalities and subsequent intramolecular Friedel–Crafts cyclization, gave a step‐ladder polymer (6) in good yields with reasonable mean average molecular weights greater than 13,000. The regioselective cyclization pattern in the α position of the naphthalene core was confirmed by a comparison of the NMR data of the polymer with those of the corresponding model ladder oligomers, 12 and 13, and also a single‐crystal structure of 13. The optical spectra of the oligomers and polymers indicated that there was little electronic interaction across the binaphthyl units. The circular dichroism spectrum of 6 exhibited a strong bisignate Cotton effect in the π–π* absorption region of the planar chromophores, which reflected the strong exciton coupling within the helical polymer chain. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 5533–5545, 2006
Synthesis and characterization of a novel biodegradable, thermoplastic polyurethane elastomer
Authors: Wang Wenshou , Ping Peng , Yu Haijun , Chen Xuesi , Jing Xiabin
Published: August 2006

A series of biodegradable, thermoplastic polyurethane elastomers poly(ϵ‐caprolactone‐co‐lactide(polyurethane [PCLA–PU] were synthesized from a random copolymer of L‐lactide (LA) and ϵ‐caprolactone (CL), hexamethylene diisocyanate, and 1,4‐butanediol. The effects of the LA/CL monomer ratio and hard‐segment content on the thermal and mechanical properties of PCLA–PUs were investigated. Gel permeation chromatography, IR, 13C NMR, and X‐ray diffraction were used to confirm the formation and structure of PCLA–PUs. Through differential scanning calorimetry, tensile testing, and tensile‐recovery testing, their thermal and mechanical properties were characterized. Their glass‐transition temperatures were below −8 °C, and their soft domains became amorphous as the LA content increased. They displayed excellent mechanical properties, such as a tensile strength as high as 38 MPa, a tensile modulus as low as 10 MPa, and an elongation at break of 1300%. Therefore, they could find applications in biomedical fields, such as soft‐tissue engineering and artificial skin. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 5505–5512, 2006
Exfoliation of montmorillonite by the insertion of disklike micelles via the soap‐free emulsion polymerization of methyl methacrylate
Authors: Lin King‐Fu , Lin Shanq‐Chyang , Chien An‐Ting , Hsieh Chi‐Chun , Yen Ming‐Huei , Lee Chia‐Hsin , Lin Chao‐Sung , Chiu Wen‐Yen , Lee Yuan‐Hon
Published: August 2006

Montmorillonites (MMTs) intercalating a potassium persulfate initiator were readily exfoliated in the presence of polymerizing methyl methacrylate during soap‐free emulsion polymerization. Because of the confined space, the polymerizing chains were aggregated into a disk form inside the clay interlayer regions for micellation. It was surprising to find from X‐ray diffraction analysis that the exfoliation of MMT was almost completed in the micellation stage. Further polymerization afforded latex particles of exfoliated poly(methyl methacrylate) (PMMA)–MMT nanocomposites. In addition, two kinds of MMT nanosheets were collected by the removal of PMMA with toluene: One was a single‐layer nanosheet with a thickness of 1.25 nm, and the other was a stacked sheet with an average thickness of 7.5 nm that may have consisted of six individual layers. Electron diffraction analysis revealed that if the individual layer of an MMT nanosheet were not a single crystal, it should contain an ultrathin layer of ice with a commonly occurring hexagonal phase. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 5572–5579, 2006
Multicyclic polyethers by the polycondensation of 1,2‐ or 1,3‐dicyanotetrafluorobenzene with flexible diphenols
Authors: Kricheldorf Hans R. , Schellenberg Johann , Schwarz Gert
Published: August 2006

1,2‐Dicyanotetrafluorobenzene (1,2‐DCTB) was polycondensed with various flexible diphenols in a molar ratio of 1:2, and experimental parameters such as the concentration and temperature were varied. Certain diphenols allowed a complete substitution of all CF bonds, so perfect multicyclic polyethers (BnCN, where B stands for bridge units, C represents cycles, and N is the degree of polymerization) were the main reaction products. Despite complete conversion, gelation was avoidable under optimized reaction conditions. However, in the case of 1,3‐dicyanotetrafluorobenzene (1,3‐DCTB), complete tetrasubstitution was not feasible with a feed ratio of 1:2. Yet, because of the inductive and mesomeric electronic interactions of all substituents in 1,3‐DCTB, the three CF groups in the ortho position with respect to the cyano groups were significantly more reactive than the fourth CF bond. Therefore, polycondensations with diphenols in a 3:2 feed ratio showed a relatively clean course, yielding soluble multicycles of structure Bn /2CN. All the multicyclic polyethers were amorphous and possessed molar mass distributions with polydispersities greater than 2. Heating with Cu2+ salts caused crosslinking of the multicycles derived from 1,2‐DCTB because of the formation of phthalocyanine complexes. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 5546–5556, 2006