bluuebunny/crossref_metadata_2025 huggingface dataset:
Mutation in G6PD gene leads to loss of cellular control of protein glutathionylation: Mechanism and implication
Authors: Ayene Iraimoudi S. , Biaglow John E. , Kachur Alexander V. , Stamato Thomas D. , Koch Cameron J.
Published: May 2007

More than 400 million people are susceptible to oxidative stress due to glucose‐6‐phosphate dehydrogenase (G6PD) deficiency. Protein glutathionylation is believed to be responsible for loss of protein function and/or cellular signaling during oxidative stress. To elucidate the implications of G6PD deficiency specifically in cellular control of protein glutathionylation, we used hydroxyethyldisulfide (HEDS), an oxidant which undergoes disulfide exchange with existing thiols. G6PD deficient (E89) cells treated with HEDS showed a significant increase in protein glutathionylation compared to wild‐type (K1) cells. In order to determine whether increase in global protein glutathionylation by HEDS leads to loss of function of an important protein, we compared the effect of HEDS on global protein glutathionylation with that of Ku protein function, a multifunctional DNA repair protein, using a novel ELISA. E89 cells treated with HEDS showed a significant loss of Ku protein binding to DNA. Cellular protein thiol and GSH, whose disulfide is involved in protein glutathionylation, were decreased by HEDS in E89 cells with no significant effect in K1 cells. E89 cells showed lower detoxification of HEDS, that is, conversion of disulfide HEDS to free sulfhydryl mercaptoethanol (ME), compared to K1 cells. K1 cells maintained their NADH level in the presence of HEDS but that of E89 cells decreased by tenfold following a similar exposure. NADPH, a cofactor required to maintain reduced form of the thiols, was decreased more in E89 than K1 cells. The specific role of G6PD in the control of such global protein glutathionylation and Ku function was further demonstrated by reintroducing the G6PD gene into E89 (A1A) cells, which showed a normal phenotype. J. Cell. Biochem. 103: 123–135, 2008. © 2007 Wiley‐Liss, Inc.
2,3,7,8‐tetrachlorodibenzo‐p‐dioxin increases bovine herpesvirus type‐1 (BHV‐1) replication in madin‐darby bovine kidney (MDBK) cells in vitro
Authors: Fiorito F. , Pagnini U. , De Martino L. , Montagnaro S. , Ciarcia R. , Florio S. , Pacilio M. , Fucito A. , Rossi A. , Iovane G. , Giordano A.
Published: May 2007

Dioxin—2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD) is a common environmental toxin of current interest. In the last years, higher levels of TCDD than those permitted in UE [European Commission. 2002. European Commission Recommendation 2002/201/CE. Official Gazette, L 67/69] were detected in milk samples from cow, water buffalo, goat, and sheep raised on some areas of Campania Region (South Italy). Dioxin often causes immunosuppression and might render the animal liable to viral infections. In addition, viral infections are able to alter the pattern of dioxin distribution in different organs of the exposed animals. Bovine Herpesvirus type‐1 (BHV‐1) is a widespread pathogen, which causes infectious rhinotracheitis and infectious pustular vulvovaginitis in cattle. Herein, we have studied the effects of TCDD and BHV‐1 infection, in Madin‐Darby Bovine Kidney (MDBK) cells, alone as well as in association, so as cellular proliferation, apoptosis, and virus replication. We have observed an increase in cell viability of confluent monolayers at low TCDD concentrations. TCDD treated cells demonstrated increased viability compared to controls as evaluated by MTT test. TCDD exposure increased cell proliferation but induced no changes on apoptosis. Cells exposed to TCDD along with BHV‐1 showed a dose‐dependent increase in cytopathy, represented by ample syncytia formation with the elimination of the cellular sheets and increased viral titer. These results suggest that TCDD increases viral replication in MDBK cells while BHV‐1 further decreases viability of TCDD exposed cells. Since very low concentrations (0.01 pg/ml) are sufficient to augment BHV‐1 titer, TCDD may contribute to reactivate BHV‐1 from latency, leading to recurrent disease and increase virus transmission. J. Cell. Biochem. 103: 221–233, 2008. © 2007 Wiley‐Liss, Inc.
The structure of the TGF‐β latency associated peptide region determines the ability of the proprotein convertase furin to cleave TGF‐βs
Authors: Kusakabe Makoto , Cheong Pak‐Leng , Nikfar Reza , McLennan Ian S. , Koishi Kyoko
Published: May 2007

The TGF‐β family members are generated as latent pre‐pro‐polypeptides. The active mature peptides are cleaved from the latent forms by cellular proteases. TGF‐β1, for instance, is predominantly processed by a substilisin‐like proprotein convertase, furin. TGF‐β2 has a consensus cleavage site for furin and therefore has been presumed to be cleaved by furin. However, TGF‐β2 is often secreted as the latent form, which appears to be inconsistent with its postulated sensitivity to furin. We report here that both the regular (short) form of TGF‐β2 and its spliced variant with an additional exon (long form) are insensitive to furin. NIH 3T3 and CHO cells were transfected with expression vectors containing the short or long form of TGF‐β2 or a chimeric TGF‐β consisting of the TGF‐β1 LAP region, the TGF‐β2 cleavage site and the TGF‐β2 mature peptide. The constructs included a c‐myc epitope tag in the N‐terminal region of the mature peptide. The TGF‐βs produced by the transfected cells were analyzed with Western blots and immunocytochemistry. The intracellular proteins harvested from these cells were incubated with furin. Furin only inefficiently cleaved both the long and short forms of TGF‐β2, but efficiently processed the chimeric TGF‐β. This indicates that the insensitivity of both forms of TGF‐β2 to furin is a consequence of the tertiary structure of their LAP regions rather than their cleavage site. This differential processing of TGF‐β1 and ‐β2 may be part of the mechanism that generates isoform‐specific functions of the TGF‐βs. J. Cell. Biochem. 103: 311–320, 2008. © 2007 Wiley‐Liss, Inc.
Rapamycin inhibits osteoblast proliferation and differentiation in MC3T3‐E1 cells and primary mouse bone marrow stromal cells
Authors: Singha Ujjal K. , Jiang Yu , Yu Shibing , Luo Min , Lu Yi , Zhang Jian , Xiao Guozhi
Published: May 2007

While the roles of the mammalian target of rapamycin (mTOR) signaling in regulation of cell growth, proliferation, and survival have been well documented in various cell types, its actions in osteoblasts are poorly understood. In this study, we determined the effects of rapamycin, a specific inhibitor of mTOR, on osteoblast proliferation and differentiation using MC3T3‐E1 preosteoblastic cells (MC‐4) and primary mouse bone marrow stromal cells (BMSCs). Rapamycin significantly inhibited proliferation in both MC‐4 cells and BMSCs at a concentration as low as 0.1 nM. Western blot analysis shows that rapamycin treatment markedly reduced levels of cyclin A and D1 protein in both cell types. In differentiating osteoblasts, rapamycin dramatically reduced osteoblast‐specific osteocalcin (Ocn), bone sialoprotein (Bsp), and osterix (Osx) mRNA expression, ALP activity, and mineralization capacity. However, the drug treatment had no effect on osteoblast differentiation parameters when the cells were completely differentiated. Importantly, rapamycin markedly reduced levels of Runx2 protein in both proliferating and differentiating but not differentiated osteoblasts. Finally, overexpression of S6K in COS‐7 cells significantly increased levels of Runx2 protein and Runx2 activity. Taken together, our studies demonstrate that mTOR signaling affects osteoblast functions by targeting osteoblast proliferation and the early stage of osteoblast differentiation. J. Cell. Biochem. 103: 434–446, 2008. © 2007 Wiley‐Liss, Inc.
Expanding the lysyl hydroxylase toolbox: New insights into the localization and activities of lysyl hydroxylase 3 (LH3)
Authors: Myllylä Raili , Wang Chunguang , Heikkinen Jari , Juffer Andre , Lampela Outi , Risteli Maija , Ruotsalainen Heli , Salo Antti , Sipilä Laura
Published: May 2007

Hydroxylysine and its glycosylated forms, galactosylhydroxylysine and glucosylgalactosylhydroxylysine, are post‐translational modifications unique to collagenous sequences. They are found in collagens and in many proteins having a collagenous domain in their structure. Since the last published reviews, significant new data have accumulated regarding these modifications. One of the lysyl hydroxylase isoforms, lysyl hydroxylase 3 (LH3), has been shown to possess three catalytic activities required sequentially to produce hydroxylysine and its glycosylated forms, that is, the lysyl hydroxylase (LH), galactosyltransferase (GT), and glucosyltransferase (GGT) activities. Studies on mouse models have revealed the importance of these different activities of LH3 in vivo. LH3 is the main molecule responsible for GGT activity in mouse embryos. A lack of this activity causes intracellular accumulation of type IV collagen, which disrupts the formation of basement membranes (BMs) during mouse embryogenesis and leads to embryonic lethality. The specific inactivation of the LH activity of LH3 causes minor alterations in the structure of the BM and collagen fibril organization, but does not affect the lifespan of mutated mice. Recent data from zebrafish demonstrate that growth cone migration depends critically on the LH3 glycosyltransferase domain. LH3 is located in the ER loosely associated with the membranes, but, unlike the other isoforms, LH3 is also found in the extracellular space in some tissues. LH3 is able to adjust the amount of hydroxylysine and hydroxylysine‐linked carbohydrates of extracellular proteins in their native conformation, suggesting that it may have a role in matrix remodeling. J. Cell. Physiol. 212: 323–329, 2007. © 2007 Wiley‐Liss, Inc.
Retracted: Novel approaches for gene‐specific interference via manipulating actions of microRNAs: Examination on the pacemaker channel genes HCN2 and HCN4
Authors: Xiao Jiening , Yang Baofeng , Lin Huixian , Lu Yanjie , Luo Xiaobin , Wang Zhiguo
Published: May 2007

Recent evidence has suggested microRNAs as viable therapeutic targets for a wide range of human disease. However, lack of gene‐specificity of microRNA actions may hinder this application. Here we developed two new approaches, the gene‐specific microRNA mimic and microRNA‐masking antisense approaches, to explore the possibility of using microRNA's principle of actions in a gene‐specific manner. We examined the value of these strategies as rational approaches to develop heart rate‐reducing agents and “biological pacemakers” by manipulating the expression of the cardiac pacemaker channel genes HCN2 and HCN4. We showed that the gene‐specific microRNA mimics, 22‐nt RNAs designed to target the 3′untranslated regions (3′UTRs) of HCN2 and HCN4, respectively, were efficient in abrogating expression and function of HCN2 and HCN4. The gene‐specific microRNA mimics repressed protein levels, accompanied by depressed f‐channel conductance and the associated rhythmic activity, without affecting mRNA levels of HCN2 and HCN4. Meanwhile, we also designed the microRNA‐masking antisense based on the miR‐1 and miR‐133 target sites in the 3′UTRs of HCN2 and HCN4 and found that these antisense oligodeoxynucleotides markedly enhanced HCN2/HCN4 expression and function, as reflected by increased protein levels of HCN2/HCN4 and If conductance, by removing the repression of HCN2/HCN4 expression induced by endogenous miR‐1/miR‐133. The experimental examination of these techniques and the resultant findings not only indicate feasibility of interfering miRNA action in a gene‐specific fashion but also may provide a new research tool for studying function of miRNAs. The new approaches also have the potential of becoming alternative gene therapy strategies. J. Cell. Physiol. 212: 285–292, 2007. © 2007 Wiley‐Liss, Inc.
Cytoplasm‐localized SIRT1 enhances apoptosis
Authors: Jin Qihuang , Yan Tingting , Ge Xinjian , Sun Cheng , Shi Xianglin , Zhai Qiwei
Published: May 2007

In general, SIRT1 is localized in nuclei. Here, we showed that endogenous and exogenous SIRT1 were both able to partially localize in cytoplasm in certain cell lines, and cytoplasm‐localized SIRT1 was associated with apoptosis and led to increased sensitivity to apoptosis. Furthermore, we demonstrated that translocation of nucleus‐localized SIRT1 from nuclei to cytoplasm was the main pathway leading to localization of SIRT1 in cytoplasm. In HeLa cells, wild type SIRT1 was completely localized in nuclei. By truncation of two predicted nuclear localization signals or fusion with an exogenous nuclear export signal, SIRT1 was partially localized in cytoplasm of HeLa cells and resulted in increased sensitivity to apoptosis. The apoptosis enhanced by cytoplasm‐localized SIRT1 was independent of its deacetylase activity, but dependent on caspases. SIRT1 was distributed in cytoplasm at metaphase during mitosis, and overexpression of SIRT1 significantly augmented apoptosis for cells at metaphase. In summary, we found SIRT1 is able to localize in cytoplasm, and cytoplasm‐localized SIRT1 enhances apoptosis. J. Cell. Physiol. 213: 88–97, 2007. © 2007 Wiley‐Liss, Inc.
Increased Ca2+ storage capacity of the skeletal muscle sarcoplasmic reticulum of transgenic mice over‐expressing membrane bound calcium binding protein junctate
Authors: Divet Alexandra , Paesante Silvia , Grasso Cristiano , Cavagna Dario , Tiveron Cecilia , Paolini Cecilia , Protasi Feliciano , Huchet‐Cadiou Corinne , Treves Susan , Zorzato Francesco
Published: May 2007

Junctate is an integral sarco(endo)plasmic reticulum protein expressed in many tissues including heart and skeletal muscle. Because of its localization and biochemical characteristics, junctate is deemed to participate in the regulation of the intracellular Ca2+ concentration. However, its physiological function in muscle cells has not been investigated yet. In this study we examined the effects of junctate over‐expression by generating a transgenic mouse model which over‐expresses junctate in skeletal muscle. Our results demonstrate that junctate over‐expression induced a significant increase in SR Ca2+ storage capacity which was paralleled by an increased 4‐chloro‐m‐cresol and caffeine‐induced Ca2+ release, whereas it did not affect SR Ca2+‐dependent ATPase activity and SR Ca2+ loading rates. In addition, junctate over‐expression did not affect the expression levels of SR Ca2+ binding proteins such as calsequestrin, calreticulin and sarcalumenin. These findings suggest that junctate over‐expression is associated with an increase in the SR Ca2+ storage capacity and releasable Ca2+ content and support a physiological role for junctate in intracellular Ca2+ homeostasis. J. Cell. Physiol. 213: 464–474, 2007. © 2007 Wiley‐Liss, Inc.
Dystroglycan expression is reduced during prostate tumorigenesis and is regulated by androgens in prostate cancer cells
Authors: Sgambato A. , De Paola B. , Migaldi M. , Di Salvatore M. , Rettino A. , Rossi G. , Faraglia B. , Boninsegna A. , Maiorana A. , Cittadini A.
Published: May 2007

Prostate cancer, the most frequently diagnosed cancer in Western men, can display a high variability in term of clinical aggressiveness and prognosis and none of the available markers is able to accurately predict its clinical course. Dystroglycan (DG), a non‐integrin adhesion molecule, is a complex formed by two subunits, α‐ and β‐DG, which bind to extracellular matrix molecules and cytoskeleton, respectively. DG expression is frequently reduced in human cancers and has been related to tumor grade and aggressiveness. This study investigated the role of DG in human prostate tumorigenesis and its suitability as a prognostic marker. The expression level of extracellular α‐DG subunit was frequently reduced in human prostate cancer cell lines and primary tumors and the percentage of positive tumor cells was significantly further decreased in vivo following androgen ablation therapy (median = 1%) compared to pre‐treatment samples (median = 28%). A significant relationship was observed between α‐DG staining on the post‐treatment samples and tumor recurrence. A dose‐ and time‐dependent decrease of DG expression also occurred in human prostate cancer cells following treatment with the anti‐androgen flutamide. Stable expression of an exogenous DG cDNA in the LNCaP human prostate carcinoma cell line resulted in a marked inhibition of both anchorage‐dependent and independent growth and of the in vivo tumorigenicity. These findings confirm and extend previous evidence that disturbances in the function of the DG complex might contribute to the definition of the malignant behavior of prostate cancer cells and suggest that androgens might regulate DG expression in these cells. J. Cell. Physiol. 213: 528–539, 2007. © 2007 Wiley‐Liss, Inc.
Mechanisms of uremic erythrocyte‐induced adhesion of human monocytes to cultured endothelial cells
Authors: Pandolfi Assunta , Di Pietro Natalia , Sirolli Vittorio , Giardinelli Annalisa , Di Silvestre Sara , Amoroso Luigi , Di Tomo Pamela , Capani Fabio , Consoli Agostino , Bonomini Mario
Published: May 2007

In end‐stage renal disease (ESRD) endothelium may represent a key target for the action of circulating elements, such as modified erythrocytes (RBC) and/or plasmatic factors, that may facilitate inflammation and the vasculopathy associated with uremia. We have previously demonstrated that phosphatidylserine (PS) exposure on the surface of RBC from ESRD patients increases RBC‐human umbilical vein endothelial cell (HUVEC) interactions and causes decreased nitric oxide (NO) production. We postulated that, besides the pro‐inflammatory effects due to decreased NO bio‐availability, enhanced ESRD‐RBC‐HUVEC interactions might directly stimulate pro‐inflammatory pathways leading to increased vascular adhesion molecule expression. ESRD‐RBC‐endothelial cell interactions induced a time‐dependent up‐regulation of VCAM‐1 and ICAM‐1 (measured by Western blot (WB) and real‐time PCR), associated with mitogen‐activated protein kinase (MAPK) activation and impairment of the Akt/endothelial nitric oxide synthase (eNOS) signaling cascade, measured by WB. In reconstitution experiments, normal RBC incubated with uremic plasma showed increased PS exposure and significantly increased VCAM‐1 and ICAM‐1 mRNA levels when incubated on HUVEC. Interestingly, ESRD‐RBC induced increased expression of adhesion molecules was prevented by Annexin‐V (AnV, able to mask PS on RBC surface), anti‐integrin‐αvβ3, anti‐thrombospondin‐1 (TSP‐1), and PD98059 (a selective inhibitor of MAPK phosphorylation). Moreover, AnV reversed the ESRD‐RBC effects on MAPK and Akt/eNOS signaling pathways. Our data demonstrate that, possibly via a direct interaction with the endothelial thrombospondin‐(αvβ3) integrin complex, ESRD‐RBC‐HUVEC adhesion induces a vascular inflammatory phenotype. Thus, intervention targeting ESRD‐RBC increased adhesion to endothelium and/or MAPK and Akt/eNOS pathways may have the potential to prevent vascular lesions under uremic conditions. J. Cell. Physiol. 213:699–709. © 2007 Wiley‐Liss, Inc.
Bone morphogenetic protein 4 induces epithelial‐mesenchymal transition through MSX2 induction on pancreatic cancer cell line
Authors: Hamada Shin , Satoh Kennichi , Hirota Morihisa , Kimura Kenji , Kanno Atsushi , Masamune Atsushi , Shimosegawa Tooru
Published: May 2007

In our study, we found that bone morphogenetic protein 4 (BMP4) has a novel effect as an inducer of epithelial‐mesenchymal transition (EMT) on Panc‐1 cells, a human pancreatic carcinoma cell line. BMP4‐treated Panc‐1 cells showed loose cell contacts and a scattered, fibroblast‐like appearance along with E‐cadherin downregulation, Vimentin upregulation and enhanced cell migration, which are characteristic of EMT. BMP4 treatment also induced homeobox gene MSX2 expression, which we previously showed to be associated with EMT in pancreatic carcinoma cells. BMP4 treatment activated the Smad signaling pathway, and extracellular signal‐related kinase (ERK) and p38 mitogen‐activated kinase (MAPK) pathways in these cells. MSX2 was markedly induced by BMP4 through the ERK and p38 MAPK pathways in collaboration with the Smad signaling pathway. The repression of E‐cadherin, induction of Vimentin and enhanced cell migration disappeared when siRNA‐based MSX2 downregulated pancreatic cancer cells were treated with BMP4. These findings indicate that BMP4 may be involved in pancreatic carcinoma development through the promotion of EMT and that MSX2 is indispensable to this process. J. Cell. Physiol. 213:768–774. © 2007 Wiley‐Liss, Inc.
Deer antler regeneration: Cells, concepts, and controversies
Authors: Kierdorf Uwe , Kierdorf Horst , Szuwart Thomas
Published: May 2007

The periodic replacement of antlers is an exceptional regenerative process in mammals, which in general are unable to regenerate complete body appendages. Antler regeneration has traditionally been viewed as an epimorphic process closely resembling limb regeneration in urodele amphibians, and the terminology of the latter process has also been applied to antler regeneration. More recent studies, however, showed that, unlike urodele limb regeneration, antler regeneration does not involve cell dedifferentiation and the formation of a blastema from these dedifferentiated cells. Rather, these studies suggest that antler regeneration is a stem‐cell‐based process that depends on the periodic activation of, presumably neural‐crest‐derived, periosteal stem cells of the distal pedicle. The evidence for this hypothesis is reviewed and as a result, a new concept of antler regeneration as a process of stem‐cell‐based epimorphic regeneration is proposed that does not involve cell dedifferentiation or transdifferentiation. Antler regeneration illustrates that extensive appendage regeneration in a postnatal mammal can be achieved by a developmental process that differs in several fundamental aspects from limb regeneration in urodeles. J. Morphol., 2007 © 2007 Wiley‐Liss, Inc.
Association Between HLA‐DRB1 polymorphism, high‐risk HPV infection and cervical neoplasia in southern Chinese
Authors: Chan Paul K.S. , Cheung Tak‐Hong , Lin C. K. , Siu Shing‐Shun N. , Yim So‐Fan , Lo Keith W.K. , Cheung Jo L. K. , Tam Ann O.Y. , Tang Julian W.
Published: May 2007

Multiple determinants are involved in the progression of human papillomavirus (HPV)‐infected cervical lesion to invasive cancer. Human leukocyte antigen (HLA) polymorphism seems to play a role. This study examined the association between HLA‐DRB1 polymorphism, high‐risk HPV infection, and the development of cervical neoplasia in southern Chinese. Three hundred and seventy women with cervical neoplasia (43 cervical intraepithelial neoplasia grade II, 154 grade III, and 173 invasive cancers) and 323 controls were recruited for HLA‐DRB1 typing by a sequence‐based approach. Cervical specimens were collected for HPV detection by a consensus primer‐based polymerase chain reaction, and with the type of HPV identified by hybridization with type‐specific oligonucleotide probes. A protective effect of HLA‐DRB1*12 for cervical neoplasia was observed, and with stronger associations when subgroup analyses were carried out for patients infected with HPV16 and HPV58. The protective effect of HLA‐DRB1*13 that had been reported from other populations was not observed. The data obtained in this study showed that HLA‐DRB1*03 conferred a higher risk for HPV18‐infected, but not for HPV16‐, HPV52‐, or HPV58‐infected cervical lesions. Although, HPV52 was reported as uncommon worldwide, it was found to be the second most prevalent type in the southern Chinese population. However, no additional risk association was observed when subgroup analyses were performed for HPV52‐infected patients. The current study shows that, among southern Chinese, the outcome of HPV‐infected cervical lesions is associated with HLA‐DRB1 polymorphism. These associations often vary with the type of HPV infection. J. Med. Virol. 79:970‐976, 2007. © 2007 Wiley‐Liss, Inc.
Anti‐HTLV‐1 tax antibody and tax‐specific cytotoxic T lymphocyte are associated with a reduction in HTLV‐1 proviral load in asymptomatic carriers
Authors: Akimoto Masaki , Kozako Tomohiro , Sawada Takashi , Matsushita Kakushi , Ozaki Atsuo , Hamada Heiichiro , Kawada Hideaki , Yoshimitsu Makoto , Tokunaga Masahito , Haraguchi Koichi , Uozumi Kimiharu , Arima Naomichi , Tei Chuwa
Published: May 2007

Previous studies have suggested that higher anti‐human T‐lymphotropic virus 1 (HTLV‐1) antibody titer and lower anti‐HTLV‐1 Tax antibody reactivity are risk factors for adult T‐cell leukemia/lymphoma. In the present study, we analyzed the relationships between these factors and clarified their significance. Forty‐five carriers were examined for anti‐HTLV‐1 and anti‐Tax antibody by ELISA. In addition, 43 of the 45 carriers with HLA‐A*0201 and/or A*2402 were examined for frequency of Tax‐specific cytotoxic T lymphocytes (CTLs) using HTLV‐1/HLA tetramers, and 44 were examined for proviral load by real‐time PCR. The relationships between these factors were analyzed statistically. The frequencies of Tax11‐19 and Tax301‐309‐specific CTLs were significantly higher in the anti‐Tax antibody‐positive group as compared with the antibody‐negative group (P = 0.002 and 0.033, respectively). Anti‐HTLV‐1 antibody titer had a positive correlation with proviral load (P = 0.019), whereas anti‐Tax antibody did not show a significant correlation. Higher frequencies of both Tax11‐19 and Tax301‐309‐specific CTLs are related to a reduction in proviral load (P = 0.017 and 0.015, respectively). Synergistic interactions of humoral and cellular immunity against Tax protein were demonstrated in HTLV‐1 carriers. Tax‐specific CTL may reduce HTLV‐1 proviral load to prevent asymptomatic carriers from developing adult T‐cell leukemia/lymphoma. J. Med. Virol. 79:977‐986, 2007. © Wiley‐Liss, Inc.
Molecular characterization of hepatitis A virus isolates from Argentina
Authors: Munné María S. , Vladimirsky Sara , Otegui Lucio , Soto Sonia , Brajterman Leonardo , Castro Raúl , Velasco María C. Cañero , Bonnano Alicia , Fernández Eduardo , Remondegui Carlos , Passeggi Carlos , Rodríguez Claudia , Pizarro Marta , Fabre Adriana , Moreiro Rita , Quarleri Jorge , González Jorge E.
Published: May 2007

Hepatitis A, a vaccine preventable disease, is now of transitional or intermediate endemicity in Argentina, as the epidemiologic pattern of the disease has shifted with improvements in living conditions in some parts of the country. Increase in the susceptibility of older children and adults has led to increasing disease incidence. Molecular epidemiology has played an important role in the understanding of HAV infection by identifying modes of spreading and by permitting the monitoring of changes in circulating virus brought about by prevention programs. South American isolates characterized are limited. Eighty‐two sporadic and outbreak isolates from Argentina were sequenced in the VP1/2A region of HAV genome over a 9‐year period. All the isolates belonged to subgenotype IA. All our sequences grouped into two big clusters. Apparently, at least two lineages have been co‐circulating in the same place at the same time. Despite great genetic variability, few point amino acid changes could be deduced. Four sequences showed an Arg → Lys substitution at 1–297 which characterized the genotype IB at the amino acid level. Many isolates carried a conservative amino acid substitution Leu → Ile at position 42 of the 2A domain, previously described as a possible fingerprint of HAV sequences in Brazil. The other rare changes have been found before, except for a 1–277 Asn → Ser substitution displayed in two isolates that has not been previously reported. Argentina recently implemented universal vaccination in 1‐year‐old children. Molecular tools would be useful in an active surveillance program. J. Med. Virol. 79:887–894, 2007. © 2007 Wiley‐Liss, Inc.
Genotyping of recent measles virus strains from Russia and Vietnam by nucleotide‐specific multiplex PCR
Authors: Kremer Jacques R. , Nguyen Giang H. , Shulga Sergey V. , Nguyen Phuc H. , Nguyen Ut T. , Tikhonova Nina T. , Muller Claude P.
Published: May 2007

The nucleoprotein genes of 49 measles virus (MV) strains circulating in Russia between 2000 and 2006 and in Vietnam in 2003 were analyzed by genotype‐specific PCR and the results were compared with their sequences. The sequences revealed the presence of genotypes H1 and H2 in the center (Nha Trang) and the north (Hanoi) of Vietnam, respectively. The relative diversity of the H2 strains suggested an endemic circulation of these viruses in the capital. In contrast genotype H1 strains from Nha Trang were homogenous genetically, which may indicate a recent importation. The strains obtained from 12 different regions of the Russian Federation were assigned to the genotypes H1, D4, and D6. Most strains (81.4%) were correctly genotyped by a multiplex PCR method which was sensitive to genotype‐specific mutations [Kremer et al. (2004): J Clin Microbiol 42: 3017–3022]. Ambiguous or negative results for some clade H and genotype D6 strains were due to point mutations in the type‐specific primer binding sites. After exchanging a single nucleotide in both the clade H‐ and the genotype D6‐specific primers, all strains were assigned correctly to their genotype. A simplified procedure for use in Vietnam was developed to distinguish directly between genotypes H1 and H2 and any non‐H genotype. These results demonstrate that our multiplex PCR method can be adapted easily to new sequence variants or specific epidemiological situations, and thus be very useful for rapid genotyping of large number of samples even in laboratories which do not have sequencing facilities. J. Med. Virol. 79:987‐994, 2007. © 2007 Wiley‐Liss, Inc.
Application of molecular and serological assays to case based investigations of rubella and congenital rubella syndrome
Authors: Jin Li , Thomas Brenda
Published: May 2007

Rubella and congenital rubella syndrome continue to be important health problems worldwide. The detection of rubella RNA directly in clinical specimens is a critical factor in early laboratory diagnosis of recent or congenital infection, in addition to detection of rubella‐specific IgM. In order to comply with recent WHO recommendations for establishing uniform genetic analysis protocols for rubella virus we have developed a new block based PCR assay (PCR‐E317), which extends the sequence generated by the block based PCR‐E592 currently in use, to cover the minimum acceptable 739 nucleotides (nt) window at the E1 gene. In addition, a real‐time PCR assay has been developed to allow rapid detection of the virus in the laboratory. The assays were applied to a number of clinical specimens collected from patients including recent rubella incidences in the UK, Ethiopia and Turkey, two prenatal and two congenital rubella syndrome cases. Rubella RNA was detected in specimens from two patients that were collected too early for IgM detection, in two amniotic fluids for prenatal diagnosis and in the follow up specimens from the two infant with congenital rubella syndrome tested for viral secretion. At least four genotypes were identified among these patients. The results showed that molecular assays are important tools in the early diagnosis of rubella and congenital rubella syndrome, in the provision of molecular epidemiological information for tracking transmission pathways and in adding to the knowledge of rubella strain distribution worldwide. J. Med. Virol. 79:1017‐1024, 2007. © 2007 Wiley‐Liss, Inc.
Seroepidemiology of hepatitis E virus in patients with non‐A, non‐B, non‐C hepatitis in Hungary
Authors: Haagsman Annika , Reuter Gábor , Duizer Erwin , Nagy Gyuláné , Herremans Tineke , Koopmans Marion , Szücs György
Published: May 2007

Many cases of acute hepatitis remain undiagnosed and the hepatitis E virus (HEV) is emerging in industrialized countries. The aim of this study was to assess the role HEV as causative agent in acute non‐A, non‐B, and non‐C hepatitis patients in Hungary. 10.5% of the 264 acute non‐A, non‐B, and non‐C hepatitis patients tested had anti‐HEV IgG and 1.9% had anti‐HEV IgM as tested by ELISA. After confirmation by Western blot 6.1% of the acute non‐A, non‐B, and non‐C hepatitis patients had anti‐HEV IgG antibodies only and 1.1% of the patients had both IgG and IgM. All 19 patients that were positive for anti‐HEV IgG and/or IgM tested negative for HEV RNA by PCR. Only a small proportion of the acute hepatitis cases in the southwest of Hungary are assumed to be attributed to HEV infection, however, hepatitis E should be considered along with hepatitis A, B, and C in the diagnosis of acute hepatitis. J. Med. Virol. 79:927–930, 2007. © 2007 Wiley‐Liss, Inc.
Different genotype pattern of varicella‐zoster virus obtained from patients with varicella and zoster in Germany
Authors: Sauerbrei A. , Wutzler P.
Published: May 2007

The general use of the varicella vaccine requires the surveillance of varicella‐zoster virus (VZV) strains in patients infected with VZV. This paper reports the data achieved from a prospective study of genotyping VZV in Germany, analyzing the restriction fragment length polymorphism (RFLP) of the open reading frames (ORF) 38, 54, and 62 as well as the polymorphism of the R5 repeat region. The study included 177 patients with varicella. Seventy‐eight patients with zoster served as controls. Results revealed that 78% of VZV strains in patients with varicella had the genetic profile of the dominant wild‐genotype occurring in Europe and 22% had the markers of African or Asian strains. Varicella patients with the profile of African or Asian strains were significantly younger than patients with varicella caused by the dominant genotype. By contrast, all zoster patients exhibited strains representing the majority of wild‐type strains in Europe. In conclusion, VZV strains from patients with varicella have a significantly higher genetic variability than viral strains from zoster patients. Since variants with the markers of African or Asian strains could only be found in young children with chickenpox, the results suggest a changing scene of VZV genotypes in Germany. As reasons, the spread of viruses, which may be imported originally by persons immigrating from warmer climates, or the recombination between wild‐and vaccine‐type viruses have to be considered. J. Med. Virol. 79:1025‐1031, 2007. © 2007 Wiley‐Liss, Inc.
EBNA1 expression in a lung transplant recipient with hypocomplementemic urticarial vasculitis syndrome
Authors: Berggren Malin Å. M. , Heinlen Latisha , Isaksson Åsa , Nyström Ulla , Ricksten Anne
Published: May 2007

This article describes a transplant recipient with underlying hypocomplementemic urticarial vasculitis syndrome who expressed persistently Epstein‐Barr virus nuclear antigen 1 (EBNA1) in peripheral blood. The patient received a bilateral lung transplant and was subsequently followed with monitoring of EBV expression in peripheral blood. Evaluation of viral expression in peripheral blood, serum, and graft tissue was performed with RT‐PCR, Q‐PCR, indirect immunofluorescence, anti‐peptide assays, and in situ hybridization; samples were collected at various time‐points up to 91 days post‐transplantation. The patient expressed EBNA1 in 8/10 (80%) of the peripheral blood samples tested during the post‐transplantation period, and interestingly, even including the day of transplantation. After analyses of indicative EBV mRNA, EBNA1 expression was found mainly to be Qp‐initiated EBNA1, known to be important for EBV maintenance. Anti‐EBNA1 epitope mapping showed significantly higher and broader antibody responses to EBNA1 epitopes pre‐transplantation when compared to normal controls and a matched lung transplant control. Post‐transplantation this response was largely diminished but there were still epitopes significantly higher than controls. Our results show the presence of EBV‐positive proliferating cells before onset of intensive immunosuppressive treatment. Although no previous connection between EBV and hypocomplementemic urticarial vasculitis syndrome has been reported, it is tempting to speculate that the continuous EBNA1 expression is not caused by immunosuppression or post‐transplant lymphoproliferative disease, but may be a factor involved in the etiology of the autoimmune disease. J. Med. Virol. 79:963‐969, 2007. © 2007 Wiley‐Liss, Inc.
Effective program against mother‐to‐child transmission of HIV at Saint Camille Medical Centre in Burkina Faso
Authors: Simpore J. , Pietra V. , Pignatelli S. , Karou D. , Nadembega W.M.C. , Ilboudo D. , Ceccherini‐Silberstein F. , Ghilat‐Avoid‐Belem W.N. , Bellocchi M.C. , Saleri N. , Sanou M.J. , Ouedraogo C.M. , Nikiema J.B. , Colizzi V. , Perno C.P. , Castelli F. , Musumeci S.
Published: May 2007

The present research was aimed to prevent mother‐to‐child transmission of HIV; to use RT‐PCR in order to detect, 6 months after birth, infected children; and to test the antiretroviral resistance of both children and mothers in order to offer them a suitable therapy. At the Saint Camille Medical Centre, 3,127 pregnant women (aged 15–44 years) accepted to be enrolled in the mother‐to‐child transmission prevention protocol that envisages: (i) Voluntary Counselling and Testing for all the pregnant women; (ii) Antiretroviral therapy for HIV positive pregnant women and for their newborns; (iii) either powdered milk feeding or short breast‐feeding and RT‐PCR test for their children; (iv) finally, pol gene sequencing and antiretroviral resistance identifications among HIV positive mothers and children. Among the patients, 227/3,127 HIV seropositive women were found: 221/227 HIV‐1, 4/227 HIV‐2, and 2/227 mixed HIV infections. The RT‐PCR test allowed the detection of 3/213 (1.4%) HIV infected children: 0/109 (0%) from mothers under ARV therapy and 3/104 (2.8%) from mothers treated with Nevirapine. All children had recombinant HIV‐1 strain (CRF06_CPX) with: minor PR mutations (M36I, K20I) and RT mutations (R211K). Among them, two twins had Non‐Nucleoside Reverse Transcriptase Inhibitor mutation (Y18CY). Both mothers acquired a major PR mutation (V8IV), investigated 6 months after a single‐dose of Nevirapine. Prevention by single‐dose of Nevirapine reduced significantly mother‐to‐child transmission of HIV, but caused many mutations and resistance to antiretroviral drugs. Based on present study the antiretroviral therapy protocol, together with the artificial‐feeding, might represent the ideal strategy to avoid transmission of HIV from mother‐to‐child. J. Med. Virol. 79:873–879, 2007. © 2007 Wiley‐Liss, Inc.
Cellular HIV‐1 DNA quantitation in patients during simplification therapy with protease inhibitor‐sparing regimens
Authors: Sarmati Loredana , Parisi Saverio Giuseppe , Nicastri Emanuele , d'Ettorre Gabriella , Andreoni Carolina , Dori Luca , Gatti Francesca , Montano Marco , Buonomini Anna Rita , Boldrin Caterina , Palù Giorgio , Vullo Vincenzo , Andreoni Massimo
Published: May 2007

Simplified regimens containing protease‐inhibitors (PI)‐sparing combinations were used in patients with virological suppression after prolonged highly active antiretroviral therapy. This study evaluated the total HIV‐1 DNA quantitation as a predictor of long‐term success for PI‐sparing simplified therapy. Sixty‐two patients were enrolled in a prospective non‐randomized cohort. All patients have been receiving a triple‐therapy regimen, two nucleoside reverse transcriptase inhibitors (NRTIs) plus one PI, for at least 9 months and were characterized by undetectable plasma HIV‐1 RNA levels (<50 cp/ml) for at least 6 months. Patients were changed to a simplified PI‐sparing regimen to overcome PI‐associated adverse effects. HIV‐DNA levels in peripheral blood mononuclear cells (PBMCs) were evaluated at baseline and at the end of follow‐up. Patients with proviral DNA levels below the median value (226 copies/106 PBMCs) had a significant higher CD4 cell count at nadir (P = 0.003) and at enrolment (P = 0.001) with respect to patients with HIV‐DNA levels above the median value. At month 18, 53 out of 62 (85%) patients on simplified regimen showed virological success, 4 (6.4%) patients experienced virological failure and 5 (8%) patients showed viral blip. At logistic regression analysis, HIV‐DNA levels below 226 copies/106 PBMCs at baseline were associated independently to a reduced risk of virological failure or viral blip during simplified therapy (OR 0.002, 95% CI 0.001–0.46, P = 0.025). The substitution of PI with NRTI or non‐NRTIs may represent an effective treatment option. Indeed, treatment failure or viral blip were experienced by 6% and 8% of the patients on simplified therapy, respectively. In addition, sustained suppression of the plasma viral load was significantly correlated with low levels of proviral DNA before treatment simplification. J. Med. Virol. 79:880–886, 2007. © 2007 Wiley‐Liss, Inc.
Characteristics of lamivudine‐resistant hepatitis B virus (HBV) strains with and without breakthrough hepatitis in patients with chronic hepatitis B evaluated by serial HBV full‐genome sequences
Authors: Horiike Norio , Duong Tran Nhu , Michitaka Kojiro , Joko Kouji , Hiasa Yoichi , Konishi Ichiro , Yano Makoto , Onji Morikazu
Published: May 2007

Lamivudine therapy often causes breakthrough of hepatitis B virus (HBV) DNA and breakthrough hepatitis. The aim of this study was to determine the viral factors that relate to HBV‐DNA breakthrough with and without breakthrough hepatitis. Among 82 patients with chronic hepatitis B (CHB) who received lamivudine at a dose of 100 mg daily for more than 24 months, 23 patients had HBV‐DNA breakthrough induced by a lamivudine‐resistant mutant. Of these 23 patients, 16 had breakthrough hepatitis and 7 had only HBV‐DNA breakthrough. Serial HBV‐DNA full‐genome sequences during therapy were examined in 10 (7 had breakthrough hepatitis and 3 did not) of these 23 patients by direct sequencing. Mutations in the S region were examined by cloning in representative patients. There were no significant differences in the baseline clinical backgrounds and virus marker between patients with and without breakthrough hepatitis. The HBV amino acid substitutions at breakthrough hepatitis were identical to those at HBV‐DNA breakthrough. Cloning analysis revealed that monoclonal mutational strain appeared at breakthrough and no such mutations existed at baseline. Regarding HBV amino acid substitutions in the polymerase region, S region, X region, and precore‐core region with breakthrough compared to baseline, there was no significant differences of the numbers of amino acid substitution between breakthrough hepatitis and non‐breakthrough hepatitis. There were no common amino acid changes in patients with breakthrough hepatitis. Although monoclonal lamivudine‐resistant strain emerged at HBV‐DNA breakthrough in patients with CHB, there were no common amino acid changes, suggesting viral factor may have insignificant role in breakthrough hepatitis. J. Med. Virol. 79:911–918, 2007. © 2007 Wiley‐Liss, Inc.
Does serum alpha interferon measurement aid in the etiological diagnosis of febrile adult patients?
Authors: Juillien Gaëlle , Haroche Julien , Bonnet Pascale , Rozenberg Flore , Riou Bruno , Hausfater Pierre
Published: May 2007

In febrile patients, distinguishing bacterial from viral infections is crucial for early treatment initiation and rational use of antibiotics. Raised interferon‐alpha (IFN‐α) levels in serum has been associated with a wide range of viral infections. We evaluated the effectiveness of IFN‐α serum measurements for the etiological diagnosis of febrile patients. Adult patients who were attending the emergency department with body temperature above or equal to 38.5°C were studied prospectively, followed‐up until day 30, and classified by two independent experts (blind for IFN‐α results) as having a bacterial/parasitic infection, viral infection, or other diagnosis. The results of IFN‐α measurements in blood samples taken in the emergency room, were compared with expert diagnosis. Among 243 patients included, 167 had bacterial/parasitic infections (including 19 with viral co‐infection), 59 had viral infections, and 36 other diagnoses. IFN‐α assay had a sensitivity of 0.44 and a specificity of 0.92 for the diagnosis of viral infection. Among the 20 patients with acute viral infection according to the emergency physician diagnosis, 7 (35%) were given antibiotics, including four patients with raised IFN‐α concentrations. It is concluded that in febrile patients, raised serum IFN‐α level is highly specific of the viral etiology of fever but poorly sensitive. Reliable viral and bacterial biological markers are needed in order to improve rational use of antibiotics. J. Med. Virol. 79:935–938, 2007. © 2007 Wiley‐Liss, Inc.
Effect of repetition rate on the formation of microtears in tendon in an in vivo cyclical loading model
Authors: Nakama Leena H. , King Karen B. , Abrahamsson Sven , Rempel David M.
Published: May 2007

We reported previously the formation of microtears in an in vivo loaded Flexor Digitorum Profundus (FDP) rabbit tendon with a repetition rate of 60 repetitions per minute and a peak force of 15% of maximum peak tetanic force for 80 cumulative hours. Tear area as a percent of tendon area, tear density (tears/mm2), and mean tear size (µm2) were higher in tendons from the loaded limb compared to the unloaded control limb. The purpose of the present study was to compare those results to results obtained with a repetition rate of 10 while maintaining the same peak force and force‐time integral (n = 8). Due to a strain gradient between the inner and outer sides of the FDP tendon, microtears were quantified in four regions, two regions each along the inner and outer sides of the tendon. The tear area as a percent of total tendon area and the mean tear size were significantly greater in the loaded limb compared to the unloaded limb (p < 0.03). However, the effects were less than those observed at 60 repetitions/min. The higher repetition loading pattern resulted in an increase in tear measures in all four regions, while the lower rate produced changes only in the outer regions of the tendon. This finding may establish where the initial sites of damage occur in tendons that insert into bone in a similar arrangement as the FDP. The results suggest that repetition rate or number of loading cycles is associated with increased tendon microtears or fragility in a dose‐response pattern. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:1176–1184, 2007
Weak Covering at Large Cardinals
Authors: Schindler Ralf ‐ Dieter
Published: May 2007

We show that weakly compact cardinals are the smallest large cardinals k where k+ < k+ is impossible provided 0# does not exist. We also show that if k+Kc < k+ for some k being weakly compact (where Kc is the countably complete core model below one strong cardinal), then there is a transitive set M with M ⊨ ZFC + “there is a strong cardinal”.
Powerset Residuated Algebras and Generalized Lambek Calculus
Authors: Kolowska‐Gawiejnowicz Miroslawa
Published: May 2007

We prove a representation theorem for (abstract) residuated algebras: each residuated algebra is isomorphically embeddable into a powerset residuated algebra. As a consequence, we obtain a completeness theorem for the Generalized Lambek Calculus. We use a Labelled Deductive System which generalizes the one used by Buszkowski [4] and Pankrat'ev [17] in completeness theorems for the Lambek Calculus.