bluuebunny/crossref_metadata_2025 huggingface dataset:
Association of lamivudine‐resistant mutational patterns with the antiviral effect of adefovir in patients with chronic hepatitis B
Authors: Cha Choong Keun , Kwon Hyeok Choon , Cheong Jae Youn , Cho Sung Won , Hong Sun Pyo , Kim Soo‐Ok , Yoo Wang Don
Published: January 2009

Adefovir has a potent antiviral activity as a rescue treatment against lamivudine‐resistant strains. The aim of this study was to assess the patterns of lamivudine‐resistant mutations and their influence on the virologic response to adefovir rescue therapy in patients with lamivudine‐resistant chronic hepatitis B. Sixty‐seven patients with lamivudine‐resistant chronic hepatitis B were treated with adefovir monotherapy. Baseline blood samples were analyzed for lamivudine‐resistant mutations via restriction fragment mass polymorphism. Virologic responses, ALT normalization and loss of HBeAg were assessed. Serum HBV DNA levels were measured using real‐time PCR at baseline and 24 weeks of adefovir therapy. Of the 67 patients with chronic hepatitis B, 65 patients (97%) had lamivudine‐resistant mutations in the YMDD motif [27 (41%) rtM204I, 22 (34%) rtM204V, and 16 (25%) rtM204I/V]. In addition to the YMDD mutations, the rtL180M, rtL80I, and rtV173L mutations were also present in 78%, 43%, and 11% of patients, respectively. The rtM204V mutation always accompanied rtL180M, and rtL80I was always observed in conjunction with rtM204I. Decrease in mean serum HBV did not differ between patients carrying the rtM204I versus rtM204V mutant at week 24 (−3.3 vs. −3.3 log10 copies/ml, respectively; P = 0.303). The presence of the rtL180M, rtL80I, and rtV173L did not significantly affect viral load reduction during adefovir administration. These results demonstrate that the rtL80I mutant is co‐selected with rtM204I as a compensatory mutation in the same manner as rtL180M with rtM204V, and that adefovir shows similar antiviral efficacy against all of the evaluated patterns of lamivudine‐resistant HBV mutations. J. Med. Virol. 81:417–424, 2009. © 2009 Wiley‐Liss, Inc.
Cell cycle perturbation in the hepatocytes of HCV core transgenic mice following common bile duct ligation is associated with enhanced p21 expression
Authors: Chang Ming‐Ling , Chen Tsung‐Hsing , Chang Ming‐Yu , Yeh Chau‐Ting
Published: January 2009

Hepatitis C virus (HCV) core protein has been reported to alter the cell cycle in vitro, but the data remain inconclusive, and in vitro experiments do not represent precisely events that occur in vivo, which may involve hepatic inflammation or regeneration. A group of double‐transgenic mice carrying tetracycline transactivator (tTA) and HCV core that express conditionally the HCV core in the mature liver, and single‐transgenic mice carrying only tTA were subjected to sham laparotomy, 43% partial hepatectomy, or common bile duct ligation. The cell cycle markers, including cyclin A, B1, D, E1, Mcm‐2, phosphorylated histone 3 protein, Ki67, and p21Waf1/Cip1/Sdi1 (p21), were evaluated in liver samples obtained 3 days after the operation. No significant differences in the levels of any markers were observed between the double‐ and single‐transgenic mice following sham laparotomy. Among the mice that underwent common bile duct ligation, the double‐transgenic mice had lower levels of Ki67 (P = 0.0001), higher levels of cyclin D1 (P = 0.0001), and higher levels of p21 expression than the single‐transgenic mice. Among those that underwent partial hepatectomy, the double‐transgenic mice had higher p21 expression levels, but no significant differences in the levels of any other markers were observed between the double‐ and single‐transgenic mice. It is concluded that the HCV core alters the hepatocyte cell cycle in addition to inducing G1 arrest in vivo after common bile duct ligation, and is associated with enhanced p21 expression. This may account at least partially for the strong association between HCV‐related hepatocarcinogenesis and hepatic inflammation/ fibrosis. J. Med. Virol. 81:467–472, 2009. © 2009 Wiley‐Liss, Inc.
Interaction between human papillomavirus type 5 E2 and polo‐like kinase 1
Authors: Wang Wei‐Sheng , Lee Moon‐Sing , Tseng Chih‐En , Liao I‐Huan , Huang Shu‐Ping , Lin Ru‐Inn , Li Chin
Published: January 2009

The E2 protein of the papillomavirus plays an essential role in the viral life cycle. Through a yeast two‐hybrid screening, human polo‐like kinase 1 was found to interact with human papillomavirus type 5 E2. Further characterization identified that the domains responsible for the interaction are the transactivation domain of HPV‐5 E2 and the sequence between the kinase and the polo box domains of Plk1. In vivo, Plk1 and HPV‐5 E2 are colocalized at the nuclear speckles. In the skin epithelium not infected with epidermodysplasia verruciformis associated HPVs, Plk1 is expressed in the stratum basale, indicating that the Plk1–HPV‐5 E2 interaction likely occurs in the keratinocytes at the basal layer of the epithelium upon infection of HPV‐5. Both HPV‐5 E2 and Plk1 also interact with the E2 binding domain of Brd4. The E2 binding domain of Brd4 is phosphorylated by Plk1 in vitro, and this phosphorylation event is blocked by the presence of HPV‐5 E2. Hence, these findings suggest the possibility that the cellular function of Brd4 is de‐regulated by forming a complex with HPV‐5 E2 in the infected epithelial cells. J. Med. Virol. 81:536–544, 2009. © 2009 Wiley‐Liss, Inc.
Ultrasensitive assessment of residual HIV viraemia in HAART‐treated patients with persistently undetectable plasma HIV‐RNA: A cross‐sectional evaluation
Authors: Bonora Stefano , Nicastri Emanuele , Calcagno Andrea , Gonzalez de Requena Daniel , D'Ettorre Gabriella , Sarmati Loredana , Palmisano Lucia , Vullo Vincenzo , Di Perri Giovanni , Andreoni Massimo
Published: January 2009

Improvements in HIV‐RNA assays have made accurate detection of as few as 2 copies/ml possible. This study objective was the evaluation of ultrasensitive HIV‐RNA quantitation (beneath current threshold: 50 copies/ml) in patients receiving different antiretroviral regimens. A cross‐sectional, ultrasensitive measurement of HIV‐RNA levels (detection limit: 2.5 HIV‐RNA copies/ml) was performed in 154 HIV‐1‐infected patients receiving ARV therapy, all classed as full responders according to the 50 copies/ml cut‐off. Patients were undergoing treatment with two nucleoside/nucleotide reverse transcriptase inhibitors (N/NtRTIs) plus nevirapine (NVP, n = 48), efavirenz (EFV, n = 57) or lopinavir/ritonavir (LPV/r, n = 49). Undetectable HIV‐RNA (<2.5 copies/ml) occurred in 29/48 (60.4%), 24/57 (42.1%) and 14/49 (28.6%) NVP, EFV and LPV/r recipients, respectively. Mean virological‐suppression (<50 copies/ml) duration was 28.6 months (median = 22, SD = 17.8), and only in LPV/r recipients length of suppression was associated with significantly lower HIV‐RNA levels (P = 0.015). Mean nadir CD4+ cell count of 270 cells/mm3 (median = 240, SD = 194.5) was significantly lower in the LPV/r arm (P < 0.001). Nadir CD4+ level correlated with virological suppression but had opposite trends between NVP (positive) and LPV/r (negative; two tailed P = 0.01). Logistic regression analysis showed NVP was the only independent factor associated with virologic suppression. NVP has demonstrated a distinct virological advantage at sub‐clinical viral loads, possibly due to its greater penetration in extra‐vascular compartments, warranting further investigation in the context of persistent low‐level viraemia in long‐term HAART. J. Med. Virol. 81:400–405, 2009. © 2009 Wiley‐Liss, Inc.
Mutations in the interferon sensitivity‐determining region of hepatitis C virus genotype 2a correlate with response to pegylated‐interferon‐alpha 2a monotherapy
Authors: Hayashi Kazuhiko , Katano Yoshiaki , Honda Takashi , Ishigami Masatoshi , Itoh Akihiro , Hirooka Yoshiki , Nakano Isao , Urano Fumihiro , Yoshioka Kentaro , Toyoda Hidenori , Kumada Takashi , Goto Hidemi
Published: January 2009

The interferon sensitivity‐determining region (ISDR) is thought to be inhibited by the double‐stranded RNA‐dependent protein kinase (PKR). Several studies have reported a relationship between the ISDR and interferon (IFN) responsiveness. However, this relationship is controversial. The aim of this study was to investigate whether genomic heterogeneity of the ISDR among patients with hepatitis C virus (HCV) genotype 2a affects the response to pegylated‐IFN‐alpha 2a monotherapy. Eighty patients (47 men, 33 women; mean age: 54.2 ± 12.9 years) infected with HCV genotype 2a were evaluated. HCV viral loads were determined by real‐time PCR. The ISDR (amino acids 2193–2228) was examined by direct sequencing. Thirty‐one patients received subcutaneous injections of pegylated‐IFN‐alpha 2a (180 µg) once weekly for 24 weeks, and 35 patients received injections for 48 weeks. Fourteen patients withdrew from treatment. Of the remaining 66 patients, 51 (77.3%) showed a sustained virologic response. Factors related to sustained virologic response on multivariate analysis were rapid virologic response (negative HCV at 4 weeks; odds ratio: 0.033; 95% confidence interval (95% CI) 0.003–0.363;P = 0.0052) and the number of mutations in the ISDR (odds ratio: 0.025; 95% CI 0.001–0.476;P = 0.0141). There were no significant differences in other factors, including sex, age, aspartate aminotransferase, alanine aminotransferase, platelet count, duration of treatment, and HCV viral load. Rapid virologic response and the ISDR sequence variations are significantly associated with response to pegylated‐IFN‐alpha 2a monotherapy in Japanese patients with HCV genotype 2a. J. Med. Virol. 81:459–466, 2009. © 2009 Wiley‐Liss, Inc.
Primary human endothelial cells support direct but not antibody‐dependent enhancement of dengue viral infection
Authors: Arévalo Maria T. , Simpson‐Haidaris Patricia J. , Kou Zhihua , Schlesinger Jacob J. , Jin Xia
Published: January 2009

Microvascular plasma leakage is the hallmark of dengue hemorrhagic fever and dengue shock syndrome. The precise molecular mechanisms leading to microvascular leakage are yet to be determined, but dengue virus (DENV) infection and consequent endothelial cell death has been suggested as its major cause. However, the extent of endothelial cell permissiveness to DENV infection and the magnitude of cell death following DENV infection are controversial. To clarify this issue, we analyzed the kinetics and consequences of DENV infection of human umbilical vein endothelial cells (HUVEC) using a novel molecularly cloned DENV2‐16681 virus. Viral replication was detected as early as 24 hr post‐infection by RT‐PCR and plaque assays. However, merely 2% of HUVEC were DENV antigen‐positive even after 96 hr of infection as measured by the FACS indirect immunofluorescence assays. Unlike monocytes/macrophages, HUVEC did not support antibody dependent enhancement of dengue viral infection due to a lack of FcγRI and FcγRII. Furthermore, DENV infection did not increase HUVEC apoptosis as compared to mock‐infected cells. Because in vitro only a small percentage of endothelial cells were productively infected in vitro with no significant apoptosis occurring in either infected or bystander cells, it would be important to re‐examine whether direct dengue viral infection of endothelium is the major cause of the extensive vascular leakage observed in patients with dengue hemorrhagic fever and dengue shock syndrome. J. Med. Virol. 81:519–528, 2009. © 2009 Wiley‐Liss, Inc.
Detection of human bocavirus in respiratory, fecal, and blood samples by real‐time PCR
Authors: Tozer Sarah J. , Lambert Stephen B. , Whiley David M. , Bialasiewicz Seweryn , Lyon Michael J. , Nissen Michael D. , Sloots Theo P.
Published: January 2009

Human bocavirus (HBoV) has been detected worldwide in respiratory samples. Two real‐time PCR assays, targeting the non‐structural protein (NP‐1) and viral protein (VP‐1) genes, were designed and validated to detect HBoV in patients with respiratory disease, gastroenteritis, or systemic illness. Sensitivity of the NP‐1 and VP‐1 assays were equal to the conventional PCR assay previously described by Allander et al. [2005: Proc Natl Acad Sci USA 102: 12891–12896] being 100%, and giving specificity of 94% and 93%, respectively. There was no cross‐reaction identified with unrelated respiratory agents, or to human DNA. The limits of detection were 10 copies of genomic DNA equivalents per reaction for both assays. The assays were used to screen three different sample populations, combined nose, and throat swabs (n = 96) from children with acute respiratory disease, fecal samples (n = 375) from adults, and children with gastroenteritis and whole blood (n = 229) collected from 31 immunocompromised children taken over an 18‐month period. In total 17 (18%) respiratory samples and 18 (4.8%) fecal samples were identified as having HBoV present. Of the pediatric whole blood specimens investigated, HBoV was detected in six (2.6%) samples from four patients. In summary, two real‐time PCR assays targeting different genes were designed and validated for use as screening methods for the detection of HBoV. HBoV was found in three different specimen types: parent‐collected combined nose–throat swabs, fecal samples collected from symptomatic individuals and whole blood from immunocompromised children. J. Med. Virol. 81:488–493, 2009. © 2009 Wiley‐Liss, Inc.
Exceptional genetic variability of hepatitis B virus indicates that Rwanda is east of an emerging African genotype E/A1 divide
Authors: Hübschen Judith M. , Mugabo Jules , Peltier Cécile Alexandra , Karasi Jean‐Claude , Sausy Aurélie , Kirpach Pierre , Arendt Vic , Muller Claude P.
Published: January 2009

In Western Africa, hepatitis B virus (HBV) genotype E predominates throughout a vast crescent spanning from Senegal to Namibia and at least to the Central African Republic to the East. Although from most of the eastern parts of sub‐Saharan Africa only limited sets of strains have been characterized, these belong predominantly to genotype A. To study how far the genotype E crescent extends to the East, a larger number of HBV strains from Rwanda were analyzed. Phylogenetic analysis of 45 S fragment sequences revealed strains of genotypes A (n = 30), D (n = 10), C (n = 4), and B (n = 1). Twelve genotype A sequences formed a new cluster clearly separated from the reference strains of the known sub‐genotypes. Thus, with four genotypes and at least six sub‐genotypes and a new cluster of genotype A strains, HBV shows an exceptional genetic variability in this small country, unprecedented in sub‐Saharan Africa. Despite this exceptional genetic variability, not a single genotype E virus was found indicating that this country does not belong to the genotype E crescent, but is east of an emerging African genotype E/A1 divide. J. Med. Virol. 81:435–440, 2009. © 2009 Wiley‐Liss, Inc.
An improved approach to identify epidemiological and phylogenetic transmission pairs of source and contact tracing of hepatitis B
Authors: Veldhuijzen Irene K. , Mes Ted H.M. , Mostert Maria C. , Niesters Hubert G.M. , Pas Suzan D. , Voermans Jolanda , de Man Robert A. , Götz Hannelore M. , van Doornum Gerard J.J. , Richardus Jan Hendrik
Published: January 2009

The transmission of infectious diseases can be traced using epidemiological and molecular information. In the current study, the congruence was assessed between sequence data of the hepatitis B virus (HBV) and epidemiological information resulting from source and contact tracing of patients seen at the Municipal Public Health Service in Rotterdam between 2002 and 2005. HBV genotypes A‐G were present in 62 acute and 334 chronic HBV patients. At the sequence level, the identical sequences of members of epidemiological transmission pairs and the rarity of such pairs provided strong support for correctness of the hypothesized transmission routes. The molecular support for epidemiological transmission pairs derived from source and contact tracing was further assessed by using topological constraints in parsimony analyses in agreement with epidemiological information, and by taking the presence of polymorphic sites of HBV within patients into account. This, in principle, allows mutations in epidemiological clusters. Of 22 epidemiological clusters, six could be refuted, four clusters received support from the molecular analysis, and support for the remaining twelve clusters was ambiguous. Two of the four epidemiological pairs that received molecular support had diverged (by 3 and 15 mutations). These results show that levels of divergence cannot be used simply as an indicator of the likelihood that groups of sequences constitute transmission pairs. Instead, to confirm or refute transmission pairs, it is necessary to assess the likelihood of a common origin of HBV variants in epidemiologically defined transmission groups relative to the HBV diversity in the local community. J. Med. Virol. 81:425–434, 2009. © 2009 Wiley‐Liss, Inc.
Genetic variability of human immunodeficiency virus‐1 in Bahia state, Northeast, Brazil: High diversity of HIV genotypes
Authors: Monteiro Joana Paixão , Alcantara Luiz Carlos Junior , de Oliveira Tulio , Oliveira Antonio Marcos , Melo Marco Antônio Gomes , Brites Carlos , Galvão‐Castro Bernardo
Published: January 2009

The HIV‐1 genetic variability in Bahia state, Brazil, was investigated. DNA samples from 229 and 213 HIV‐1‐infected individuals were analyzed using the heteroduplex mobility assay (HMA) in gag and env fragments, respectively. One hundred seventy‐five samples were characterized in both genes. Thirty‐two subtype F and BF recombinant viruses were sequenced and analyzed by phylogenetic methods. The combination of HMA and sequencing results showed that seven different HIV‐1 genotypes comprised this sample: 147 (84%) B/B, 4 (2.3%) F/F, 3 (1.7%) B/F, 1 (0.6%) F/B, 1 (0.6%) F/D, 1 (0.6%) BF/F, and 18 (10.3%) BF/B. A significant divergence was observed between these two techniques results (84.4%). This is explained by the low accuracy of the HMA for detecting recombinant viruses. These recombinants were unrelated to CRF12, while two sequences were related to CRF28 and CRF29. Nineteen BF mosaics shared the same gag breakpoint. In conclusion, the use of HMA may be inappropriate in regions where different subtypes are co‐circulating. Subtype B is the most common genotype, however, an increased prevalence (13.1%) of different BF variants and a potentially new CRF suggest that recombination is occurring frequently in Bahia. These viruses were associated with women infected heterosexually. Finally, this study identified the presence of an F/D recombinant HIV‐1 in Brazil. J. Med. Virol. 81:391–399, 2009. © 2009 Wiley‐Liss, Inc.
Evaluation of properties of chitosan as an adjuvant for inactivated influenza vaccines administered parenterally
Authors: Ghendon Yuri , Markushin Stanislav , Vasiliev Yuri , Akopova Irina , Koptiaeva Irina , Krivtsov Georgy , Borisova Olga , Ahmatova Nelly , Kurbatova Ekaterina , Mazurina Svetlana , Gervazieva Valentina
Published: January 2009

Studies on mice showed that chitosan as an adjuvant for H5 inactivated influenza vaccines administered intramuscularly enhances significantly antibody titers and protective efficiency not only against homologous influenza viruses, but also against drift variants. Chitosan adjuvanted vaccines induced high antibody titers after a single immunization and with a low dose of antigen. High antibody titers remained for at least 6 months. Chitosan adjuvanted vaccine stored at 4°C preserves its adjuvant properties for at least 8 months. Chitosan stimulates proliferative and cytotoxic activity of splenic mononuclear leukocytes in mice and promotes an increase in the numbers of CD3, CD3/NK, I‐AK (MHC II), and H‐2Db (MHC I) cells. After intramuscular immunization, chitosan did not induce IgE antibodies and antibodies against chitosan itself. Chitosan is a promising adjuvant candidate for inactivated influenza vaccines administered parenterally. J. Med. Virol. 81:494–506, 2009. © 2009 Wiley‐Liss, Inc.
High‐risk HPV types in lesions of the uterine cervix of female commercial sex workers in the Philippines
Authors: Miyashita Michiko , Agdamag Dorothy M. , Sasagawa Toshiyuki , Matsushita Kaori , Salud Lourdes Ma. , Salud Calixto O. , Saikawa Kunikazu , Leano Prisca S. , Pagcaliwagan Teresita , Acuna Jessica , Ishizaki Azumi , Kageyama Seiji , Ichimura Hiroshi
Published: January 2009

In order to prevent cervical cancer, vaccines against human papilloma virus types 16 (HPV‐16) and 18 (HPV‐18) have been implemented worldwide. However, the HPV types that cause cancer can differ according to geographical area and ethnicity. In this new era of the HPV vaccine, it is important to elucidate the prevalent HPV types in each area. Therefore, the prevalence of HPV infection and cervical abnormalities among 369 female commercial sex workers in the Philippines were examined. HPV L1 gene was amplified by polymerase chain reaction (PCR) using modified GP5+/6+ primers, and genotyping was performed by sequencing cloned PCR products. HPV DNA was detected in 211 (57.2%) women, among whom 46 HPV types were identified. HPV‐52 was most common and multiple‐type infection was observed in 44.5%. Among 56 women with abnormal cervical cytology (low‐ and high‐grade squamous intraepithelial lesions and adenocarcinoma in situ), HPV‐52 was most common (23.2%), followed by HPV‐16 (19.6%), ‐58 (10.7%), and ‐67 (10.7%). Only 27% of these women were positive for HPV‐16 and ‐18. Multivariate analysis revealed that HPV‐16, ‐39, ‐52, ‐67, and ‐82 were significantly associated with abnormal cytology. Repeated analysis of HPV‐52 single‐positive samples using the original GP5+/6+ PCR primers produced negative results in 57% of cases, suggesting that the prevalence of HPV‐52 infection may have been underestimated in previous studies, and the current vaccines may not be sufficient for preventing infection and the development of premalignant lesions of the cervix in women in the Philippines. J. Med. Virol. 81:545–551, 2009. © 2009 Wiley‐Liss, Inc.
Identification of the novel KI Polyomavirus in paranasal and lung tissues
Authors: Babakir‐Mina Muhammed , Ciccozzi Massimo , Campitelli Laura , Aquaro Stefano , Coco Achille Lo , Perno Carlo Federico , Ciotti Marco
Published: January 2009

KI is a novel polyomavirus identified in the respiratory secretions of children with acute respiratory symptoms. Whether this reflects a causal role of the virus in the human respiratory disease remains to be established. To investigate the presence of KIV in the respiratory tissue, we examined 20 fresh lung cancer specimens and surrounding normal tissue along with one paranasal and one lung biopsy from two transplanted children. KIV‐VP1 gene was detected in 9/20 lung cancer patients and 2/2 transplanted patients. However, amplification of the sequence coding for the C‐terminal part of the early region of KIV performed on the 11 positive cases was successful only in two malignant lung tissues, one surrounding normal tissue, and 1/2 biopsies tested. Phylogenetic analysis performed on the early region of KIV (including the four Italian isolates), BKV and JCV revealed the presence of three distinct clades. Within the KIV clade two sub‐clades were observed. A sub‐clade A containing the four Italian strains, and a sub‐clade B comprising the Swedish and Australian isolates. Interestingly, the two Italian strains identified in normal tissue clustered together, whereas those detected in malignant tissue fell outside this cluster. In vitro studies are needed to investigate the transforming potential of KIV strains. J. Med. Virol. 81:558–561, 2009. © 2009 Wiley‐Liss, Inc.
Increased detection of HBV DNA in HBsAg‐positive and HBsAg‐negative South African HIV/AIDS patients enrolling for highly active antiretroviral therapy at a Tertiary Hospital
Authors: Lukhwareni Azwidowi , Burnett Rosemary J. , Selabe S. Gloria , Mzileni M. Olga , Mphahlele M. Jeffrey
Published: January 2009

This retrospective study investigated the prevalence of hepatitis B virus (HBV) in 192 stored sera from human immunodeficiency virus (HIV) positive South African patients initiating antiretroviral therapy (ART), and explored the implications of HBV–HIV co‐infection on laboratory diagnosis of HBV. HBV serology (HBsAg, anti‐HBs and anti‐HBc) and nested HBV PCR assays targeting the HBV polymerase gene were performed, with HBV DNA positive samples being quantified with Cobas Taqman HBV test 48 assay (Roche Diagnostics). The study found that 63% (121/192) of patients had past or present HBV infection, and 40.6% (78/192) had detectable HBV DNA. Also, 22.9% (44/192) of patients were HBsAg positive and HBV DNA positive, while 23% (34/148) of HBsAG negatives had occult HBV infections. Of the 78 HBV DNA positive samples, 62.8% had viral loads ranging from 102 to ≥108 IU/ml, and 37.2% had HBV viral loads <200 IU/ml. There was a statistically significant positive association between HBsAg‐positivity and high viral loads, with 27% (12/44) of HBsAg positives having HBV viral loads between 104 and ≥108 IU/ml, compared to only 5.9% (2/34) of HBsAg negatives (relative risk: 4.64; 95% confidence interval: 1.11, 19.35; chi‐square P‐value = 0.015). The study shows that the majority of HIV/AIDS patients initiating ART have either acute or chronic HBV infections, and further confirms that HIV remains a risk factor for occult HBV infections in South African patients as previously shown. The findings strongly support HBV screening in all HIV‐positive patients initiating ART in South Africa, considering that current ART regimens include drugs with anti‐HBV activity (e.g., lamivudine). J. Med. Virol. 81:406–412, 2009. © 2009 Wiley‐Liss, Inc.
Identification of mutations in a temperature‐sensitive mutant (tsm5) of murine cytomegalovirus using complementary genome sequencing
Authors: Timoshenko Olga , Al‐Ali Abdulaziz , Martin Brian A.B. , Sweet Clive
Published: January 2009

Identification of mutations in mutants derived chemically is a difficult and relatively random process. NimbleGen Comparative Genome Sequencing (CGS) was assessed as an inexpensive, rapid method of identifying mutations in the temperature‐sensitive mutant tsm5 of the K181 (Birmingham) variant of murine cytomegalovirus (MCMV). This genome resequencing approach requires an established genome sequence as a reference. Comparison of tsm5 and the K181 (Birmingham) variant with the published K181 (Perth) MCMV genomic sequence revealed a total of 10 synonymous and 15 non‐synonymous SNPs in tsm5 and 14 of the latter were confirmed by sequencing. Thus, while CGS cannot be relied upon to identify correctly all mutations it was helpful for identifying a large number of mutations for further investigation that could contribute to the ts phenotype of tsm5. J. Med. Virol. 81:511–518, 2009. © 2009 Wiley‐Liss, Inc.
Severity and age of rotavirus diarrhea, but not socioeconomic conditions, are associated with rotavirus seasonality in Venezuela
Authors: Schael Irene Pérez , González Rosabel , Salinas Belén
Published: January 2009

Rotavirus (RV) epidemiology presents differences between developing and developed countries among which are seasonality, age at first infection, variability of strain in circulation and severity of disease. Since, in Venezuela, we have distinct seasonal patterns of RV occurrence, we examined the epidemiological profile of RV disease associated to these differences by analyzing data from previous studies conducted in Venezuela. Data were collected from children <5 years of age with diarrhea seen in six hospitals located in five cities. Socio‐demographic and clinical characteristics of RV illness were analyzed according to RV identification by ELISA assay and the seasonal patterns of RV circulation (marked versus minimal seasonality). A total of 6,742 episodes of diarrhea (1,820 rotavirus positive and 4,922 rotavirus negative) were evaluated: 1,951 in Caracas, Cumaná and Pto. Ordaz (minimal seasonality) and 4,791 in Mérida and Valencia (marked seasonality). Mean age (months) of children with RV was 8.14 ± 6.74 and 12.21 ± 9.62 (P < 0.0001) in areas with minimal and marked seasonality, respectively. RV disease was more frequent (33% vs. 24%; OR = 1.536; 95% CI: 1.36–1.73), more severe (dehydration: 64% vs. 29%; OR = 4.436; 95% CI: 3.61–5.44) and more common in infants than in older children (79% vs. 60%; OR = 2.521; 95% CI: 2.01–3.14) in cities with minimal seasonality than in those with marked seasonality. Socioeconomic conditions were not associated with seasonality. In environments with minimal seasonality, children are infected with RV at younger ages and the disease is more severe regardless of malnutrition and poverty. J. Med. Virol. 81:562–567, 2009. © 2009 Wiley‐Liss, Inc.
Polymorphisms at GLUT1 gene are not associated with the development of TSP/HAM in Brazilian HTLV‐1 infected individuals and the discovery of a new polymorphism at GLUT1 gene
Authors: Costa Giselle Calasans Souza , Azevedo Rochele , Gadelha Sandra Rocha , Kashima Simone Haddad , Muricy Gabriel , Olavarria Viviana Nila , Covas Dimas Tadeu , Takayanagui Osvaldo Massaiti , Galvão‐Castro Bernardo , Alcantara Luiz Carlos Júnior
Published: January 2009

The development of HTLV‐1 associated clinical manifestations, such as TSP/HAM and ATLL, occur in 2–4% of the infected population and it is still unclear why this infection remains asymptomatic in most infected carriers. Recently, it has been demonstrated that HTLV uses the Glucose transporter type 1 (GLUT1) to infect T‐CD4+ lymphocytes and that single nucleotide polymorphisms (SNP) in the GLUT1 gene are associated with diabetic nephropathy in patients with diabetes mellitus in different populations. These polymorphisms could contribute to a higher GLUT1 protein expression on cellular membrane, facilitating the entry of HTLV and its transmission cell by cell. This could result in a higher provirus load and consequently in the development of TSP/HAM. To evaluate the role of GLUT1 gene polymorphisms in the development of TSP/HAM in HTLV‐1 infected individuals, the g.22999G > T, g.15339T > C and c.‐2841A > T sites were analyzed by PCR/RFLP or sequencing in 244 infected individuals and 102 normal controls. The proviral load of the HTLV‐1 infected patients was also analyzed using Real Time Quantitative PCR. Genotypic and allelic frequencies of the three sites did not differ significantly between controls and HTLV‐1 infected individuals. There was no difference in genotypic and allelic distributions among patients as to the presence or absence of HTLV‐1 associated clinic manifestations. As regards the quantification of the provirus load, we observed a significant reduction in the asymptomatic individuals compared with the oligosymptomatic and TSP/HAM individuals. These results suggest that g.22999G > T, g.15339T > C, and c.‐2841A > T SNP do not contribute to HTLV‐1 infection nor to the genetic susceptibility of TSP/HAM in Brazilian HTLV‐1 infected individuals. J. Med. Virol. 81:552–557, 2009. © 2009 Wiley‐Liss, Inc.
Occult hepatitis B in persons infected with HIV is associated with low CD4 counts and resolves during antiretroviral therapy
Authors: Cohen Stuart James W.T. , Velema Marieke , Schuurman Rob , Boucher Charles A.B. , Hoepelman Andy I.M.
Published: January 2009

Occult hepatitis B virus (HBV) is defined by the presence of plasma HBV DNA in individuals with HBV core antibodies (anti‐HBc), but without HBV surface antigen (HBsAg). The prevalence of occult HBV in HIV‐infected patients remains controversial, and the risk factors, clinical significance and effect of highly active antiretroviral therapy (HAART) are unknown. The aim of this study was to determine prevalence, risk factors, and clinical significance of occult HBV in HIV‐infected patients and to evaluate the effect of HAART. Plasma HBV DNA levels were determined in 191 HIV positive, antiretroviral naïve patients, who were anti‐HBc positive and HBsAg negative. Quantitative HBV DNA was determined using a Taqman real‐time nested PCR. Additionally, plasma HIV RNA levels, CD4 cell counts, anti‐HBs‐antibodies, anti‐HCV‐antibodies, ALT, AST, and γGT were determined. Occult HBV (a plasma HBV DNA level >50 copies/ml) was detected in 9/191 (4.7%) of the patients. Among 45 anti‐HBs‐negative patients (isolated anti‐HBc positive), the prevalence was 11.1%. Patients with occult HBV had significantly lower CD4 count compared to anti‐HBc‐positive/HBsAg negative/HBV DNA‐negative patients (105 ± 157 (median ± SD) vs. 323 ± 299 cells/mm3, P = 0.019). When HAART (including lamivudine) was initiated in the patients with occult HBV, HBV DNA was no longer detectable in any of the patients during 3 years of follow‐up. In conclusion, occult HBV was associated with low CD4 counts and may be viewed as opportunistic reactivation of HBV that resolves as a consequence of HAART induced immune reconstitution and/or the effect of lamivudine. J. Med. Virol. 81:441–445, 2009. © 2009 Wiley‐Liss, Inc.
Seasonal shifts of group A rotavirus strains as a possible mechanism of persistence in the human population
Authors: Parra Gabriel I.
Published: January 2009

This article demonstrates how the seasonal predominance of a new rotavirus strain in Asuncion, Paraguay is correlated with a wide spectrum of age groups of children infected in that given season. Therefore, this study provides new evidence to support the idea that seasonal shift of rotavirus strains is a possible mechanism used by the virus to evade herd immunity (acquired by the population due to previous infections) and, thus, ultimately persist in that population. J. Med. Virol. 81:568–571, 2009. © 2009 Wiley‐Liss, Inc.
Prognostic factors of long‐term CD4+count‐guided interruption of antiretroviral treatment
Authors: Sarmati L. , Andreoni C. , Nicastri E. , Tommasi C. , Buonomini A. , D'Ettorre G. , Corpolongo A. , Dori L. , Montano M. , Volpi A. , Narciso P. , Vullo V. , Andreoni M.
Published: January 2009

Aim of the study was to determine predictors of the duration of antiretroviral treatment interruption in patients infected with HIV. This pilot prospective, open‐label, multicenter trial comprised 62 HIV‐seropositive subjects who decided voluntarily to interrupt therapy after two or more years of successful HAART. The primary end‐point was the time to patients being free of therapy before reaching a CD4+ cell count ≤350/µl. Fifteen of 62 patients remained in treatment interruption for more than 180 days. Patients restarting therapy had higher HIV‐DNA levels (P = 0.05), were treated more frequently with NNRTI‐drugs (P = 0.02), had a shorter period of HAART (P = 0.046), and lower CD4+ cell counts after day 14 of interruption of treatment (P = 0.04). Multivariate regression analysis showed that less than 323 baseline proviral HIV‐DNA cp/106 PBMCs and more than 564 CD4 cells/µl at day 14 after interruption were associated independently with a reduced risk of restarting treatment (P = 0.041 and P = 0.012, respectively). A score based on CD4+ cell counts at nadir, at baseline, at week 2 of treatment interruption, and on baseline HIV‐DNA values can identify patients with a prolonged period free safely of treatment. J. Med. Virol. 81:481–487, 2009. © 2009 Wiley‐Liss, Inc.
Phosphothioated oligodeoxynucleotides induce nonspecific effects on neuronal cell adhesion in a growth substrate‐dependent manner
Authors: Okun Eitan , Mattson Mark P.
Published: January 2009

Synthetic phosphothioated (PTO) oligodeoxynucleotide (ODN) sequences are commonly used for a variety of applications that benefit from nuclease protection. The PTO modification is implemented mainly in antisense ODN, but also in ODN that were shown to activate members of the toll‐like receptor (TLR) family such as TLR3 (poly‐I:C), TLR8 (ssRNA), and TLR9 (CpG). Neurons are routinely plated on surfaces coated with either cationic substances such as poly‐L‐ornithine (PLO), polyethylenimine (PEI), poly‐L‐lysine or ECM components such as laminin, collagen, or fibronectin. We found that PTO‐ODN aimed at activating TLR9 induces a non‐TLR9‐specific detachment phenotype in cortical neurons plated on either laminin or PEI, but not on PLO. This phenotype was correlated with decreased viability and was partially inhibited when caspase‐3 was inhibited with Ac‐DEVD‐CMK. This finding suggests that the use of PTO‐ODN can cause nonspecific effects on cell adhesion that could compromise interpretation of data from experiments using PTO‐ODN. © 2009 Wiley‐Liss, Inc.
Monads and instanton bundles on smooth hyperquadrics
Authors: Costa Laura , Miró–Roig Rosa Maria
Published: January 2009

In this paper we study instanton bundles on quadric hypersurfaces from the algebraic and geometric point of view. We discuss examples and we give a cohomological characterization (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Accurate assessment of clinical nurses' work environments: Response rate needed
Authors: Kramer Marlene , Schmalenberg Claudia , Brewer Barbara B. , Verran Joyce A. , Keller‐Unger Jan
Published: January 2009

Improvement of hospital unit work environments is key to quality patient care, productivity, nurse retention, and job satisfaction. Accurate measurement of such environments is necessary prior to introduction and evaluation of improvement structures and strategies. Characteristics and attributes of work environments are group level phenomena. Accurate assessment of these phenomena requires survey response rates of sufficient size to ensure sample representativeness and data that can reliably be aggregated to group level. What is the sufficient response rate? This question was answered through psychometric testing of five random samples from the population of 23 M.D. Anderson Cancer Center clinical units that had 100% response rates on an environmental survey. Response rates of 40% or more had acceptable psychometric properties for unit‐specific scales. © 2009 Wiley Periodicals, Inc. Res Nurs Health 32:229–240, 2009
Lymphangiogenesis in gastric carcinoma correlates with prognosis
Authors: Gao Peng , Zhou Geng‐Yin , Zhang Qing‐Hui , Su Zhong‐Xue , Zhang Ting‐Guo , Xiang Lei , Wang Yan , Zhang Shun‐Li , Mu Kun
Published: January 2009

Lymphatic metastasis is an important way that gastric carcinomas can spread. However, little is known about the mechanisms of lymphangiogenesis and its clinical significance in gastric carcinomas. In the present study, lymphatic vessel density (LVD), VEGF‐C expression, and proliferative activity of lymphatic endothelium were determined in human gastric carcinomas and xenografts of gastric cancer cells in nude mice. The development of lymphangiogenesis and its correlation with patient prognosis were investigated. The results showed that lymphatic vessels were observed mainly in peripheral tumour tissue with significantly (p < 0.05) higher P‐LVD (peri‐tumoural‐LVD) than I‐LVD (intra‐tumoural‐LVD). The expression of VEGF‐C was heterogeneous within tumours, with a significantly higher expression (immunostaining score) at the margin than at the tumour centre (p < 0.05). A significant correlation was found between VEGF‐C expression at the margin (but not at the centre) and P‐LVD (r = 0.72, p < 0.01). High proliferative activity of lymphatic endothelium was also observed in the peripheral tissues, with a significant correlation between proliferative activity of lymphatic endothelium and VEGF‐C expression (p < 0.05). These data imply that the increased lymphatics may have been newly formed following stimulation by VEGF‐C. High VEGF‐C expression at the margin of gastric carcinomas could induce lymphangiogenesis in the peri‐tumoural stroma and contribute to the increased P‐LVD. The data from mice tumour xenografts also suggested that VEGF‐C produced from the transplanted gastric carcinoma cells could induce lymphangiogenesis around them. In patients, VEGF‐C expression at tumour margins was associated with nodal metastasis, lymphatic vessel invasion, poor recurrence‐free survival, and poor overall survival, and could serve as an independent predictor for patients with gastric carcinoma. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Retrospective analysis of relapsed or primary refractory childhood lymphoblastic lymphoma in Japan
Authors: Mitsui Tetsuo , Mori Tetsuya , Fujita Naoto , Inada Hiroko , Horibe Keizo , Tsurusawa Masahito
Published: January 2009

Background and Procedure

To assess the clinical course with response to second‐line treatment and to evaluate the role of hematopoietic stem cell transplantation (SCT) in children with relapsed or primary refractory lymphoblastic lymphoma (LBL), we analyzed data of 48 patients with relapsed/primary refractory diseases among 260 LBL patients identified in a national survey of 1996–2004.

Results

Twenty‐six patients achieved second complete remission; 9 achieved partial remission. Of 13 patients who showed progression despite first and second line therapy, only one patient was alive on the second relapse after unrelated cord blood transplantation. Among 40 relapsed patients, the median time between initial diagnosis and relapse was 12.5 months (range 3–56 months). The sites of relapse were isolated BM (n = 9), primary local site with BM (9), primary local site (6), isolated CNS (4), local site with mediastinum (4), primary local site with other site (4), and others (4). Of all 48 patients, 3 were alive after chemotherapy alone. Of the 33 patients, 14 were alive after high dose chemotherapy (HDC)/SCT. With a 27.5‐month median follow up period, the 3‐year OS rate was 43.2 ± 7.4% (estimate ± SE). Univariate analysis identified two features (relapse within 12 months, T cell phenotype) as significant variables that predicted poor survival. Multivariate analysis showed novel statistically significant variables including relapse within 12 months from initial diagnosis (Hazard ratio 3.60) and absence of HDC/SCT (2.64).

Conclusion

Outcomes of patients with relapsed/primary refractory LBL were poor, but HDC/SCT for these patients was associated with good results. Pediatr Blood Cancer 2009;52:591–595. © 2009 Wiley‐Liss, Inc.
X‐ray structure of Danio rerio secretagogin: A hexa‐EF‐hand calcium sensor
Authors: Bitto Eduard , Bingman Craig A. , Bittova Lenka , Frederick Ronnie O. , Fox Brian G. , Phillips George N.
Published: January 2009

Many essential physiological processes are regulated by the modulation of calcium concentration in the cell. The EF‐hand proteins represent a superfamily of calcium‐binding proteins involved in calcium signaling and homeostasis. Secretagogin is a hexa‐EF‐hand protein that is highly expressed in pancreatic islet of Langerhans and neuroendocrine cells and may play a role in the trafficking of secretory granules. We present the X‐ray structure of Danio rerio secretagogin, which is 73% identical to human secretagogin, in calcium‐free form at 2.1‐Å resolution. Secretagogin consists of the three globular domains each of which contains a pair of EF‐hand motifs. The domains are arranged into a V‐shaped molecule with a distinct groove formed at the interface of the domains. Comparison of the secretagogin structure with the solution structure of calcium‐loaded calbindin D28K revealed a striking difference in the spatial arrangement of their domains, which involves ∼180° rotation of the first globular domain with respect to the module formed by the remaining domains. Proteins 2009. © 2009 Wiley‐Liss, Inc.
The local structure of α‐FeSi2 under high pressure
Authors: Mori Yoshihisa , Nakano Hiroko , Sakane Genta , Aquilanti Giuliana , Udono Haruhiko , Takarabe Kenichi
Published: January 2009

We report the results of the high‐pressure X‐ray diffraction study and the high‐pressure extended X‐ray absorption fine structure study for α‐FeSi2 using the synchrotron radiation sources. The bond lengths of Fe–Fe, Si–Si, and Fe–Si under pressure were refined by the Rietveld method and the EXAFS analysis. It is interesting that the bond length of Si–Si is almost unchanged with increasing pressure. The DV‐Xα molecular orbital calculation was carried out for the crystal structure at 34.9 GPa and shows that the highest occupied molecular orbital consists of only 3d orbital of Fe atom. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Magnetic and structural properties of transition metal doped zinc‐oxide nanostructures
Authors: Ankiewicz Amélia O. , Gehlhoff Wolfgang , Martins Joana S. , Pereira Ângela S. , Pereira Sérgio , Hoffmann Axel , Kaidashev Evgeni M. , Rahm Andreas , Lorenz Michael , Grundmann Marius , Carmo Maria C. , Trindade Tito , Sobolev Nikolai A.
Published: January 2009

We report on the magnetic and structural properties of two types of nanostructures doped with Co or Mn, namely, ZnO nanowires and colloidal ZnO nanocrystals. Electron paramagnetic resonance (EPR) spectra have been measured and analysed to extract information on the incorporation of the ions in the lattice. A detailed analysis by means of simulations of the experimental EPR spectra confirms that the transition metal (TM) ions were mainly incorporated as TM2+, occupying the Zn2+ sites in the wurtzite structure of ZnO. Furthermore, for both types of nanostructures, the EPR spectra are composed of more than one signal, revealing locally distorted environments or core‐shell structures, proved by surface modifications via inorganic coatings. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Bias dependent spatially resolved photoluminescence spectroscopy and photocurrent measurements of InGaN/GaN LED structures
Authors: Vierheilig Clemens , Schwarz Ulrich T. , Gmeinwieser Nikolaus , Laubsch Ansgar , Hahn Berthold
Published: January 2009

The optical properties of Nitride‐based quantum well devices are affected by internal fields, fluctuation of the composition of the quantum well and nonradiative recombination. In this paper, we present a method to separate these effects by optical spectroscopy and photocurrent measurements under varying applied bias. We observe long‐range fluctuations both in the photoluminescence (PL) properties and the photocurrent, while fluctuations on the sub‐mm length scale can only be observed in the PL. A comparison with simplified 1D calculations leads to the assumption, that the long‐range fluctuations are caused by fluctuations of the quantum well width, while the short‐range fluctuations are explained by a varying nonradiative recombination (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Electrical and structural characterizations of BGaN thin films grown by metal‐organic vapor‐phase epitaxy
Authors: Baghdadli T. , Hamady S. Ould Saad , Gautier S. , Moudakir T. , Benyoucef B. , Ougazzaden A.
Published: January 2009

In this report, the electrical and structural properties of BGaN thin films, with boron composition up to 2%, have been investigated. The resistivity, Hall mobility and carrier concentration were measured by the van der Pauw/Hall Effect technique, using annealed indium ohmic contacts. The current‐voltage measurements between two planar gold and titanium/aluminum electrodes were performed. To analyze the structural properties, the polarized Raman spectra were measured with a micro‐Raman spectrometer. The resistivity displays a strong increase with respect to the boron composition in BGaN alloy, directly related to the decrease of the n‐type carrier concentration, while the mobility increases with boron content, well correlated to the structural measurements showing a good crystalline quality of the BGaN layers. This boron‐controlled resistivity is very promising for using the BGaN based materials in microelectronic and optoelectronic devices. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Influence of strain and polarisation on electronic properties of a GaN/AlN quantum dot
Authors: Young Toby D. , Marquardt Oliver
Published: January 2009

It is known that heterojunctions give rise to changes in the elastic and piezoelectric properties of a given semiconductor nanostructure. For practical applications it is decisive to understand the influence of these strain and piezoelectric fields on the electronic response of these systems, both as distinct and as coupled fields. In the present study, the confinement properties of the electron and hole wavefunctions are of interest. Therefore, a model based on a traditional envelope 8‐band k · p Hamiltonian is implemented and used to explore a GaN/AlN quantum dot/matrix system (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)