bluuebunny/crossref_metadata_2025 huggingface dataset:
A new whole genome amplification method for studying clonal evolution patterns in malignant colorectal polyps
Authors: Hirsch Daniela , Camps Jordi , Varma Sudhir , Kemmerling Ralf , Stapleton Mark , Ried Thomas , Gaiser Timo
Published: February 2012

To identify the genetic drivers of colorectal tumorigenesis, we applied array comparative genomic hybridization (aCGH) to 13 formalin‐fixed paraffin‐embedded (FFPE) samples of early, localized human colon adenocarcinomas arising in high‐grade adenomas (so‐called “malignant polyps”). These lesions are small and hence the amount of DNA is limited. Additionally, the quality of DNA is compromised due to the fragmentation as a consequence of formalin fixation. To overcome these problems, we optimized a newly developed isothermal whole genome amplification system (NuGEN Ovation® WGA FFPE System). Starting with 100 ng of FFPE DNA, the amplification system produced 4.01 ± 0.29 μg (mean ± standard deviation) of DNA. The excellent quality of amplified DNA was further indicated by a high signal‐to‐noise ratio and a low derivative log2 ratio spread. Both, the amount of amplified DNA and aCGH performance were independent of the age of the FFPE blocks and the associated degradation of the extracted DNA. We observed losses of chromosome arms 5q and 18q in the adenoma components of the malignant polyp samples, while the embedded early carcinomas revealed losses of 8p, 17p, and 18, and gains of 7, 13, and 20. Aberrations detected in the adenoma components were invariably maintained in the embedded carcinomas. This approach demonstrates that using isothermally whole genome amplified FFPE DNA is technically suitable for aCGH. In addition to demonstrating the clonal origin of the adenoma and carcinoma part within a malignant polyp, the gain of chromosome arm 20q was an indicator for progression from adenoma to carcinoma. Published 2012 Wiley Periodicals, Inc.
Illegitimate V(D)J recombination involving nonantigen receptor loci in lymphoid malignancy
Authors: Onozawa Masahiro , Aplan Peter D.
Published: February 2012

V(D)J recombination of antigen receptor loci (IGH, IGK, IGL, TCRA, TCRB, TCRG, and TCRD) is an essential mechanism that confers enormous diversity to the mammalian immune system. However, there are now at least six examples of intrachromosomal interstitial deletions caused by aberrant V(D)J recombination between nonantigen receptor loci; five of out these six are associated with lymphoid malignancy. The SIL‐SCL fusion and deletions of CDKN2A, IKZF1, Notch1, and Bcl11b are all associated with lymphoid malignancy. These interstitial deletions seem to be species specific, as the deletions seen in mice are not seen in humans; the converse is true as well. Nucleotide sequence analysis of these rearrangements reveals the hallmarks of V(D)J recombination, including site specificity near cryptic heptamer signal sequences, exonucleolytic “nibbling” at the junction site, and nontemplated “N”‐region nucleotide insertion at the junction site. Two of these interstitial deletions (murine Notch1 and Bcl11b deletions) have been detected, at low frequency, in tissues from healthy mice with no evidence of malignancy, similar to the finding of chromosomal translocations in the peripheral blood or tonsils of healthy individuals. The contention that these are mediated via V(D)J recombination is strengthened by in vivo assays using extrachromosomal substrates, and chromatin immunoprecipitation‐sequence analysis which shows Rag2 binding at the sites of rearrangement. Although the efficiency of these “illegitimate” recombination events is several orders of magnitude less than that at bona fide antigen receptor loci, the consequence of such deletions, namely activation of proto‐oncogenes or deletion of tumor suppressor genes, is devastating, and a major cause for lymphoid malignancy. © 2012 Wiley Periodicals, Inc.
Aging of myelinating glial cells predominantly affects lipid metabolism and immune response pathways
Authors: Verdier Valérie , Csárdi Gábor , de Preux‐Charles Anne‐Sophie , Médard Jean‐Jacques , Smit August B. , Verheijen Mark H. G. , Bergmann Sven , Chrast Roman
Published: February 2012

Both the central and the peripheral nervous systems are prone to multiple age‐dependent neurological deficits, often attributed to still unknown alterations in the function of myelinating glia. To uncover the biological processes affected in glial cells by aging, we analyzed gene expression of the Schwann cell‐rich mouse sciatic nerve at 17 time points throughout life, from day of birth until senescence. By combining these data with the gene expression data of myelin mouse mutants carrying deletions of either Pmp22, SCAP, or Lpin1, we found that the majority of age‐related transcripts were also affected in myelin mutants (54.4%) and were regulated during PNS development (59.5%), indicating a high level of overlap in implicated molecular pathways. The expression profiles in aging copied the direction of transcriptional changes observed in neuropathy models; however, they had the opposite direction when compared with PNS development. The most significantly altered biological processes in aging involved the inflammatory/immune response and lipid metabolism. Interestingly, both these pathways were comparably changed in the aging optic nerve, suggesting that similar biological processes are affected in aging of glia‐rich parts of the central and peripheral nervous systems. Our comprehensive comparison of gene expression in three distinct biological conditions including development, aging, and myelin disease thus revealed a previously unanticipated relationship among themselves and identified lipid metabolism and inflammatory/immune response pathways as potential therapeutical targets to prevent or delay so far incurable age‐related and inherited forms of neuropathies. © 2012 Wiley Periodicals, Inc.
Fatty liver index, gamma-glutamyltransferase, and early carotid plaques
Authors: Kozakova Michaela , Palombo Carlo , Eng Marco Paterni , Dekker Jacqueline , Flyvbjerg Allan , Mitrakou Asimina , Gastaldelli Amalia , Ferrannini Ele
Published: February 2012

An association between fatty liver and carotid atherosclerosis has been established; however, it is not clear whether this relationship is a consequence of shared conventional risk factors or whether it is determined by specific circulating factors originating from liver or adipose tissue. To identify the factors possibly linking fatty liver and atherosclerosis, we assessed, in 1,012 subjects free of confounding diseases (e.g., hypertension, diabetes, cardiovascular diseases, and dyslipidemia) and metabolic syndrome, the relationship between the presence of early plaques at carotid bifurcation and fatty liver index (FLI; a validated surrogate marker of fatty liver), as well as the associations between carotid plaque presence and established atherosclerotic risk factors, family history of cardiovascular disease (FH-CVD) or diabetes, insulin sensitivity, serum liver enzymes, adipokines, fatty free acids, and high-sensitivity C-reactive protein (hsCRP). A total of 55 of 1,012 subjects (5.4%) had small plaque at carotid bifurcation. Subjects with plaque were older and had higher prevalence of FLI ≥60 and FH-CVD, higher blood pressure, plasma low-density lipoprotein cholesterol, glucose, gamma-glutamyltransferase (GGT), and hsCRP, as compared to subjects without plaques (

P

< 0.05). In a logistic regression model, adjusted for sex, liver transaminase, and alcohol consumption, the independent predictors of plaque presence were age (

P

< 0.0005), FLI ≥60 (

P

< 0.0005), and current smoking (

P

< 0.05). When FLI in the model was replaced by variables used in its equation (e.g., body mass index, waist circumference, plasma triglycerides, and GGT), the independent determinants of plaque presence were age (

P

< 0.001), GGT (

P

= 0.001), and current smoking (

P

< 0.05).

Conclusions:

Our cross-sectional study suggests that subjects with FLI ≥60 are at higher risk of atherosclerotic lesions, independently of established risk factors, and that serum GGT may represent a link between fatty liver and the development of early atherosclerosis. (Hepatology 2012)
Chronic hepatitis C genotype 1 patients with insulin resistance treated with pioglitazone and peginterferon alpha-2a plus ribavirin
Authors: Harrison Stephen A. , Hamzeh Fayez M. , Han Jian , Pandya Prashant K. , Sheikh Muhammed Y. , Vierling John M.
Published: February 2012

Patients with chronic hepatitis C and insulin resistance are less likely to respond to anti-hepatitis C virus (HCV) therapy and are at risk for more rapid fibrosis progression. Coadministration of pioglitazone with peginterferon/ribavirin improves insulin sensitivity and increases virologic response rates in insulin-resistant HCV genotype 4 patients, but it is unclear whether this finding applies to genotype 1 patients. For this reason we randomized treatment-naive HCV genotype 1 patients with insulin resistance to receive either standard care (peginterferon alpha-2a plus ribavirin for 48 weeks, n = 73) or pioglitazone 30-45 mg/day plus standard care (n = 77) in an open-label multicenter trial. Patients randomized to pioglitazone received the drug during a 16-week run-in phase, the 48-week standard-care phase, and the 24-week untreated follow-up phase. Pioglitazone treatment improved hemoglobin A1c (HbA1c), plasma glucose, insulin levels, and homeostasis model assessment of insulin resistance score and increased serum adiponectin levels during the 16-week run-in phase and maintained these improvements during the standard-care phase. However, we observed no statistically significant difference between the two groups in the primary efficacy endpoint, the decrease from baseline to Week 12 of peginterferon alpha-2a/ribavirin treatment in mean log10 HCV RNA titer (−3.5 ± 1.71 and −3.7 ± 1.62 IU/mL in the pioglitazone and standard-care groups, respectively, Δ = 0.21 IU/mL,

P

= 0.4394).

Conclusion

: Treatment with pioglitazone before and during treatment with peginterferon alpha-2a plus ribavirin improved several indices of glycemic control in patients with chronic hepatitis C and insulin resistance, but did not improve virologic response rates compared with peginterferon alpha-2a plus ribavirin alone. (HEPATOLOGY 2012)
Regulatory T cells control the CD8 adaptive immune response at the time of ductal obstruction in experimental biliary atresia
Authors: Lages Celine S. , Simmons Julia , Chougnet Claire A. , Miethke Alexander G.
Published: February 2012

CD8 T-lymphocytes are effector cells of cholangiocyte injury in human and in rhesus rotavirus (RRV)-induced experimental biliary atresia (BA). Here we hypothesize that neonatal deficiency in CD25+CD4+ regulatory T cells (Tregs) leads to aberrant activation of hepatic T-lymphocytes in BA. We found that adoptive transfer of total CD4 cells, but not of CD25-depleted CD4 cells, prior to RRV inoculation reduced expansion of CD8 cells, plasma bilirubin levels, ductal inflammation, and bile duct epithelial injury at 7 days postinfection (dpi) compared with age-matched infected controls without adoptive transfer. Searching for mechanisms, we found that in vitro production of interferon-gamma (IFN-γ) by naïve CD8 cells upon polyclonal stimulation was enhanced in coculture with hepatic dendritic cells (DCs) from RRV-infected, but not with DCs from noninfected mice, which was correlated with an increased proportion of CD11b+ myeloid (m)DCs and up-regulation of the costimulatory molecule CD86 on RRV-primed DCs. Furthermore, DC-dependent T-lymphocyte activation was blocked by anti-CD86 antibody in dose-dependent fashion. Importantly, expression of CD86 on mDCs was down-regulated by Tregs in vitro, and adoptive transfer of Treg-containing CD4 cells decreased expression of CD86 on hepatic mDCs at 7 dpi. On the contrary, in mice resistant to experimental BA, CD25+ cell depletion aggravated bile duct injury at 12 dpi after RRV inoculation, as plasma bilirubin levels were elevated by >20-fold compared with nondepleted infected controls. Increased susceptibility to hepatobiliary injury in Treg-depleted mice was linked to hepatic CD8 expansion and enhanced stimulatory capacity of hepatic DCs. Conclusion: Activation of hepatic T-lymphocytes driving biliary obstruction in BA is regulated by mDCs by way of CD86-dependent costimulation and is susceptible to inhibition by Tregs. (HEPATOLOGY 2012;56:219–227)
Pegylated interferon-α induced hypoferremia is associated with the immediate response to treatment in hepatitis C
Authors: Ryan John D. , Altamura Sandro , Devitt Emma , Mullins Sarah , Lawless Matthew W. , Muckenthaler Martina U. , Crowe John
Published: February 2012

Pegylated interferon-α (PEG-IFN-α) forms an integral part of the current treatment for hepatitis C virus (HCV) infection. PEG-IFN-α suppresses HCV production by augmenting the innate antiviral immune response. Recent studies have reported the induction of hepcidin, the iron regulatory hormone, by IFN-α

in vitro

. As hepcidin plays an important role in innate immunity, we hypothesized that this finding may be of clinical relevance to HCV and investigated the changes in iron homeostasis during the first 24 hours of treatment. Blood samples were obtained from HCV patients immediately prior to and 6, 12, and 24 hours following the first dose of PEG-IFN-α/ribavirin (RBV). Samples were analyzed for hepcidin, cytokine, iron levels, and HCV viral load, and hepcidin messenger RNA (mRNA) expression was quantified in peripheral blood mononuclear cells. Hepcidin induction by IFN-α was further analyzed in cell culture. In HCV patients a single dose of PEG-IFN-α/RBV resulted in a significant increase in serum hepcidin, peaking at 12 hours, coinciding with a 50% reduction in serum iron and transferrin saturation over the 24-hour period. Patients with a ≥2 log decline in HCV viral load over the first 24 hours had significantly lower SI and TS levels at 12 and 24 hours. Moreover, 24-hour SI levels were an independent predictor of the immediate HCV viral decline, an indicator of ultimate treatment outcome. In cell culture, a direct induction of hepcidin by IFN-α was seen, controlled by the STAT3 transcription factor.

Conclusion

: Hepcidin induction occurs following the initiation of PEG-IFN-α treatment for HCV, and is mediated by way of STAT3 signaling. The subsequent hypoferremia was greatest in those with the most significant decline in viral load, identifying systemic iron withdrawal as a marker of immediate interferon-α efficacy in HCV patients. (HEPATOLOGY 2012)
Inhibition of xanthine oxidase reduces wasting and improves outcome in a rat model of cancer cachexia
Authors: Springer Jochen , Tschirner Anika , Hartman Kai , Palus Sandra , Wirth Eva K. , Ruis Silvia Busquets , Möller Nadine , von Haehling Stephan , Argiles Josep M. , Köhrle Josef , Adams Volker , Anker Stefan D. , Doehner Wolfram
Published: February 2012

Cachexia is a common co‐morbidity in cancer occurring in up to 80% of patients depending on the type of cancer. Uric acid (UA), the end‐product of the purine metabolism, is elevated in cachexia due to tissue wasting and upregulated xanthine oxidase (XO) activity. High serum UA levels indicate increased XO‐dependent production of oxygen free radicals (reactive oxygen species; ROS) and correlate with metabolic illness and poor survival. We hypothesized that XO‐inhibition might reduce inflammatory signals accounting for tissue wasting and improve survival in experimental cancer cachexia. Animals were inoculated intraperitoneally with AH‐130 hepatoma cells and treated with two XO‐inhibitors: allopurinol [Allo, low (LD) and high dose (HD) 4 and 40 mg/kg/d] and its more effective active metabolite oxypurinol (Oxy, 4 and 40 mg/kg/d) or placebo for 15 days. Weight loss and tissue wasting of both fat and lean tissue (assessed by NMR‐scanning) was reduced by both LD and HD Allo and LD‐Oxy, but not by HD‐Oxy. A robust induction of XO‐activity for generation of reactive oxygen species was seen in the placebo group (assessed by electron paramagnetic spectroscopy), which was reduced by XO‐inhibition. Increased ROS induced cytokine signaling, proteolytic activity and tissue degradation were all attenuated by XO inhibition. Survival was significantly and dose dependently improved. Food intake and spontaneous locomotor activity were higher, indicating a higher quality of life. Inhibition of XO can reduce tissue wasting and improve survival in cancer cachexia and clearly clinical studies are needed.
Antiangiogenic gene therapy with soluble VEGF‐receptors ‐1, ‐2 and ‐3 together with paclitaxel prolongs survival of mice with human ovarian carcinoma
Authors: Sopo Minna , Anttila Maarit , Sallinen Hanna , Tuppurainen Laura , Laurema Anniina , Laidinen Svetlana , Hamalainen Kirsi , Tuunanen Pasi , Koponen Jonna K. , Kosma Veli‐Matti , Heinonen Seppo , Alitalo Kari , Yla‐Herttuala Seppo
Published: February 2012

We compared effects of antiangiogenic gene therapy with a combination of soluble sVEGFR‐1, sVEGFR‐2 and sVEGFR‐3 to chemotherapy with carboplatin and paclitaxel and to antiangiogenic monoclonal anti‐VEGF‐antibody bevacizumab in an intraperitoneal ovarian cancer xenograft model in mice (n = 80). Gene therapy was also combined with chemotherapy. Therapy was initiated when sizable tumors were confirmed in magnetic resonance imaging (MRI). Adenovirus‐mediated gene transfer was performed intravenously (2 × 109 pfu), while chemotherapy and monoclonal anti‐VEGF‐antibody were dosed intraperitoneally. The study groups were as follows: AdLacZ control (n = 21); combination of AdsVEGFR‐1, ‐2 and ‐3 (n = 21); combination of AdsVEGFR‐1, ‐2, ‐3 and paclitaxel (n = 9); bevacizumab (n = 14); paclitaxel (n = 9) and carboplatin (n = 5). Effectiveness was assessed by survival time and surrogate measures such as sequential MRI, immunohistochemistry, microvessel density and tumor growth. Antiangiogenic gene therapy combined with paclitaxel significantly prolonged the mean survival of mice (25 days) compared to the controls (15 days) and all other treatment groups (p = 0.001). Bevacizumab treatment did not have any significant effect on the survival. Tumors of the mice treated by gene therapy were significantly smaller than in the control group (p = 0.021). The mean vascular density and total vascular area were also significantly smaller in the tumors of the gene therapy group (p = 0.01). These results show potential of the antiangiogenic gene therapy to improve efficacy of chemotherapy with paclitaxel and support testing of this approach in a phase I clinical trial for the treatment of ovarian cancer.
Thyroid cancer in Denmark 1943–2008, before and after iodine supplementation
Authors: Blomberg M. , Feldt‐Rasmussen U. , Andersen K.K. , Kjaer S.K.
Published: February 2012

Thyroid cancer incidence has increased worldwide during the previous decades. In this nationwide study, we aimed to identify the overall incidence of thyroid cancer in Denmark during 66 years (1943–2008) and incidences of the four main histological types of thyroid cancer from 1978 to 2008. Data were obtained from the nationwide Danish Cancer Registry, and we focused especially on the period after implementation of compulsory iodine supplementation, which was established on a national level in 2000. We calculated age‐standardized incidence rates per 100,000 person‐years, and age‐period‐cohort models were fitted to describe trends in incidence. To quantify trends in incidence over time, log‐linear Poisson models were used to estimate annual percentage change. From 1943 to 2008, 1,947 men (29%) and 4,682 women (71%) were diagnosed with thyroid cancer. The age‐standardized incidence increased in both sexes; in men from 0.41 to 1.57 per 100,000 and from 0.90 to 4.11 per 100,000 in women, corresponding to a significant average annual percentage change of 1.7 and 1.8%, respectively. The incidence increased with younger birth cohorts. The rise was almost exclusively caused by papillary carcinomas, and it was particularly present during the last decades of the study period. It cannot be ruled out that iodine supplementation may play a role for the risk of thyroid cancer, but as the strongest increase in incidence began in the years before the implementation, it is likely that improvement in diagnostic modalities increased diagnostic activity, and/or new unknown risk factors are also important contributors to the increase.
Cytotoxicity on human cells of Cry1Ab and Cry1Ac Bt insecticidal toxins alone or with a glyphosate‐based herbicide
Authors: Mesnage R. , Clair E. , Gress S. , Then C. , Székács A. , Séralini G.‐E.
Published: February 2012

The study of combined effects of pesticides represents a challenge for toxicology. In the case of the new growing generation of genetically modified (GM) plants with stacked traits, glyphosate‐based herbicides (like Roundup) residues are present in the Roundup‐tolerant edible plants (especially corns) and mixed with modified Bt insecticidal toxins that are produced by the GM plants themselves. The potential side effects of these combined pesticides on human cells are investigated in this work. Here we have tested for the very first time Cry1Ab and Cry1Ac Bt toxins (10 ppb to 100 ppm) on the human embryonic kidney cell line 293, as well as their combined actions with Roundup, within 24 h, on three biomarkers of cell death: measurements of mitochondrial succinate dehydrogenase, adenylate kinase release by membrane alterations and caspase 3/7 inductions. Cry1Ab caused cell death from 100 ppm. For Cry1Ac, under such conditions, no effects were detected. The Roundup tested alone from 1 to 20 000 ppm is necrotic and apoptotic from 50 ppm, far below agricultural dilutions (50% lethal concentration 57.5 ppm). The only measured significant combined effect was that Cry1Ab and Cry1Ac reduced caspases 3/7 activations induced by Roundup; this could delay the activation of apoptosis. There was the same tendency for the other markers. In these results, we argue that modified Bt toxins are not inert on nontarget human cells, and that they can present combined side‐effects with other residues of pesticides specific to GM plants. Copyright © 2012 John Wiley & Sons, Ltd.
Apoptosis of rat ovarian granulosa cells by 2,5‐hexanedione in vitro and its relevant gene expression
Authors: Zhang Wenchang , Huang Lei , Kong Cancan , Liu Jin , Luo Lingfeng , Huang Huiling
Published: February 2012

Although n‐hexane is toxic to the ovary, the related mechanism remains unknown. Apoptosis of rat ovarian granulosa cells is thought to be one of the important reasons for ovarian toxicity. Rat ovarian granulosa cells were exposed in vitro to different concentrations of 2,5‐hexanedione (HD, a major metabolite of n‐hexane; 0, 20, 40 and 60 mmol l−1) to observe the apoptosis, and its relevant gene expression was investigated. It was found by MTT assay that different exposure doses and different exposure time could inhibit the viability of ovarian granulosa cells. After a 12 h exposure, we found the apoptosis rate of the ovarian granulosa cells increased notably with the increase of HD dose. The HD concentration of each exposure dose could inhibit the message RNA (mRNA) expression of both bcl‐2 and xiap, and increase the mRNA expression of fasl without any influence on that of bcl‐xl, while only an HD concentration of 40 mol l−1 could raise the mRNA expression of bax. Concentrations of HD of 40 and 60 mol l−1 could inhibit the protein expression of NF‐κB, but only an HD concentration of 60 mol l−1 could improve theactivation of NF‐κB. In summary, our results indicate that HD can induce the apoptosis of ovarian granulosa cell, and the occurrence of apoptosis is related to inhibition of protein expression of NF‐κB, activation of NF‐κB, upregulation of mRNA levels of fasl and bax, and downregulation of mRNA levels of xiap and bcl‐2. Copyright © 2012 John Wiley & Sons, Ltd.
The application of tissue engineering in reversing mitomycin C‐induced ischemic conjunctiva
Authors: Peng Yi‐Jie , Pan Chien‐Yi , Hsieh Yi‐Ting , Wu Semon , Chu Hsi‐Ning , Hsu Wei‐Cherng
Published: February 2012

The purpose of this study is to evaluate the angiogenic potential of collagen‐glycosaminoglycan (CG) matrices in mitomycin C‐induced ischemic conjunctival defect, in New Zealand white rabbits. After creating a conjunctival defect at the center of ischemic conjunctiva, a CG matrix was implanted into subconjunctival space to evaluate the conjunctival reepithelialization and angiogenesis during the wound healing process. In the grafted group, the vessel count of the healed conjunctiva was substantially elevated by two fold within the initial 4 weeks and the increased vascular content originated mostly from the fornix site. The rate of conjunctival reepithelialization was not retarded in the grafted group, and the final thickness of healed conjunctiva was similar in both grafted and ungrafted groups. The histological studies revealed that the collagen matrix did not elicit pronounced inflammatory reaction and the regenerated conjunctiva showed loosely arranged collagen deposition without significant scar formation. The α SMA staining positive myofibroblasts were identified in the acute inflammatory stage and were absent, 8 weeks after implantation in both groups. The results indicated that the porous collagen scaffold was able to enhance vascularization and physiological recovery of ischemic conjunctival defect, implying a potential alternative therapy for the ischemic leaking bleb after glaucoma filtrating surgery in ophthalmic practices. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A:, 2012.
Inflammatory response induced by candidate biomaterials of an implantable microfabricated sensor
Authors: Sokolov A. , Hellerud B. C. , Johannessen E. A. , Mollnes T. E.
Published: February 2012

The implantation of synthetic medical devices is known to generate an immediate and complex material‐related inflammatory response. Consequently, 15 candidate materials for a new microfabricated sensor were investigated. A human whole blood model that permits the interaction of all the putative inflammatory systems was used. The experiments were performed by administering 500 μL of lepirudin‐anticoagulated blood in each well of a 24‐well polystyrene microtiter plate preloaded with the respective materials. The degree of inflammation was evaluated by assessing four complement activation markers, six proinflammatory cytokines, and chemokines, the expression of monocyte tissue factor (TF), as well as platelet activation. The complement system was inhibited with the C5‐inhibitor eculizumab. Three of the materials distinctly activated complement through the alternative pathway, whereas the rest of the materials were virtually inert. Notably, the same three materials induced a marked and selective expression of TF as well as the release of five of the six cytokines. All these increases were statistically significant (p < 0.05). Inhibition of complement by the C5‐inhibitor virtually abolished TF expression and markedly reduced several of the cytokines, suggesting that complement is a particularly useful tool to reveal the immediate inflammatory‐inducing properties of these biomaterials. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A:, 2012.
Osteoclast stimulatory transmembrane protein and dendritic cell–specific transmembrane protein cooperatively modulate cell–cell fusion to form osteoclasts and foreign body giant cells
Authors: Miyamoto Hiroya , Suzuki Takayuki , Miyauchi Yoshiteru , Iwasaki Ryotaro , Kobayashi Tami , Sato Yuiko , Miyamoto Kana , Hoshi Hiroko , Hashimoto Kazuaki , Yoshida Shigeyuki , Hao Wu , Mori Tomoaki , Kanagawa Hiroya , Katsuyama Eri , Fujie Atsuhiro , Morioka Hideo , Matsumoto Morio , Chiba Kazuhiro , Takeya Motohiro , Toyama Yoshiaki , Miyamoto Takeshi
Published: February 2012

Cell–cell fusion is a dynamic phenomenon promoting cytoskeletal reorganization and phenotypic changes. To characterize factors essential for fusion of macrophage lineage cells, we identified the multitransmembrane protein, osteoclast stimulatory transmembrane protein (OC-STAMP), and analyzed its function. OC-STAMP–deficient mice exhibited a complete lack of cell–cell fusion of osteoclasts and foreign body giant cells (FBGCs), both of which are macrophage-lineage multinuclear cells, although expression of dendritic cell specific transmembrane protein (DC-STAMP), which is also essential for osteoclast/FBGC fusion, was normal. Crossing OC-STAMP–overexpressing transgenic mice with OC-STAMP–deficient mice restored inhibited osteoclast and FBGC cell–cell fusion seen in OC-STAMP–deficient mice. Thus, fusogenic mechanisms in macrophage-lineage cells are regulated via OC-STAMP and DC-STAMP. © 2012 American Society for Bone and Mineral Research.
Assessment of the accuracy of MRI wall shear stress estimation using numerical simulations
Authors: Petersson Sven , Dyverfeldt Petter , Ebbers Tino
Published: February 2012

Purpose:

To investigate the accuracy of wall shear stress (WSS) estimation using MRI. Specifically, to investigate the impact of different parameters and if MRI WSS estimates are monotonically related to actual WSS.

Materials and Methods:

The accuracy of WSS estimation using methods based on phase‐contrast (PC) MRI velocity mapping, Fourier velocity encoding (FVE) and intravoxel velocity standard deviation mapping were studied using numerical simulations. The influence of spatial resolution, velocity encoding, wall segmentation, and voxel location were investigated over a range of WSS values.

Results:

WSS estimates were found to be sensitive to parameter settings in general and spatial resolution in particular. All methods underestimated WSS, except for the FVE‐based method, which instead was extremely sensitive to voxel position relative to the wall. Methods using PC‐based WSS estimation with wall segmentation showed to be accurate for low WSS, but were sensitive to segmentation errors.

Conclusion:

Even in the absence of noise and for relatively simple velocity profiles, MRI WSS estimates cannot always be assumed to be linearly or even monotonically related to actual WSS. High WSS values cannot be resolved and the estimates depend on parameter setting. Nevertheless, distinguishing areas of low and moderate WSS may be feasible. J. Magn. Reson. Imaging 2012;36:128–138. © 2012 Wiley Periodicals, Inc.
Bone marrow fat content in the elderly: A reversal of sex difference seen in younger subjects
Authors: Griffith James F. , Yeung David K.W. , Ma Heather Ting , Leung Jason Chi Shun , Kwok Timothy C.Y. , Leung Ping Chung
Published: February 2012

Purpose:

To investigate vertebral bone marrow fat content in elderly subjects related to sex, age, and bone mineral density (BMD) and relate these findings to published data in younger subjects.

Materials and Methods:

A total of 259 healthy subjects (145 females, 114 males; age range, 62–90 years) underwent proton (1H) MR spectroscopy of L3 vertebral body and BMD of the lumbar spine with results stratified according to age. Ninety age‐ and BMD‐matched subjects were selected to determine sex differences in marrow fat content and BMD.

Results:

In females, vertebral marrow fat content rose sharply between 55 and 65 years of age while in males vertebral marrow fat content rose gradually throughout life. Vertebral marrow fat content in females more than 60 years was approximately 10% higher in females than males, i.e., a reversal of sex difference reported in marrow fat content for subjects less than 60 years.

Conclusion:

Marrow fat content increases sharply in female subjects between 55 and 65 years of age while male subjects continue to increase marrow fat at a more gradual steady rate. Females older than 60 years have a higher marrow fat content than males. This increased deposition in marrow fat concurs with recognized changes in extraosseous fat distribution in postmenopausal females. J. Magn. Reson. Imaging 2012;36:225–230. © 2012 Wiley Periodicals, Inc.
The epidemiology and etiology of influenza‐like illness in Chinese children from 2008 to 2010
Authors: Peng Jinsong , Kong Wenhua , Guo Deyin , Liu Manqing , Wang Ying , Zhu Honghao , Pang Beibei , Miao Xiaoping , Yu Bin , Luo Tongyong , Hu Quan , Zhou Dunjin
Published: February 2012

Influenza‐like illness can be caused by a wide range of respiratory viruses. In order to investigate the epidemiology of viral pathogens related to influenza‐like illness in children of Wuhan, the largest city in central China, throat swab samples were collected from 1,472 young patients, from July 2008 to June 2010, before and after the occurrence of the 2009 pandemic influenza A (H1N1) virus (pH1N1). It was found that 923 patients (62.7%) were positive for at least 1 virus and 90 patients (9.8%) were detected for multiple (≥2) respiratory viruses by real‐time PCR detection of 16 viruses. Seasonal influenza A virus was the predominant pathogen among all the 16 viruses with a positive rate of 13.3% (196/1,472), which was followed by pH1N1 (159/1,472). It was also noted that the viral distribution pattern in Wuhan changed upon the introduction of the pH1N1 virus. J. Med. Virol. 84:672–678, 2012. © 2011 Wiley Periodicals, Inc.
Human parechovirus infections in patients admitted to hospital in Northern Italy, 2008–2010
Authors: Piralla Antonio , Furione Milena , Rovida Francesca , Marchi Antonietta , Stronati Mauro , Gerna Giuseppe , Baldanti Fausto
Published: February 2012

Human parechoviruses (HPeVs) infection is associated with a wide range of clinical syndromes such as respiratory, gastrointestinal, neurologic diseases, and neonatal sepsis‐like illness. The main objective of this study was to investigate the epidemiology of HPeVs infection in hospitalized patients in a period of 2 years. Respiratory samples from 3,525 patients with respiratory syndrome, cerebrospinal fluid (CSF) from 340 patients with neurologic syndrome as well as CSF and plasma samples from five neonatal patients with sepsis‐like illness collected from October 2008 to 2010 were tested retrospectively using HPeV‐specific real‐time RT‐PCR. Phylogenetic analysis of VP3/VP1 region was performed on the positive samples. Fourteen out of 3,525 (0.4%) patients with respiratory syndrome and five out of five patients with sepsis‐like illness were positive for HPeV. In 3/5 patients with sepsis‐like illness multiple samples (e.g., stool, plasma, CSF, or respiratory samples) were available, and HPeV was found in all specimens. In contrast, no positive CSF was detected among the 340 patients with neurologic syndromes. Eleven patients (57.9%) were infected with HPeV1 strain, 7 (36.8%) with HPeV3, and 1 (5.3%) with HPeV6 strains. Ten of the 14 HPeV patients with respiratory syndrome were co‐infected with other respiratory viruses (eight with rhinovirus and two with coronavirus OC43). All five patients with sepsis‐like illness were less than 1 month of age and were infected with HPeV3. Although not circulating at high frequency and unlikely to cause respiratory syndrome, HPeV was associated with severe clinical syndromes in a minority of newborns. J. Med. Virol. 84:686–690, 2012. © 2011 Wiley Periodicals, Inc.
Introduction of new subtypes and variants of hepatitis C virus genotype 4 in South Africa
Authors: Gededzha Maemu P. , Selabe Selokela G. , Kyaw Thanda , Rakgole J. Nare , Blackard Jason T. , Mphahlele M. Jeffery
Published: February 2012

Hepatitis C virus (HCV) genotype is an important predictor of disease progression and treatment response. This descriptive study investigated the sequence diversity and genotypes of HCV in South Africa based on comparative analysis of the 5′ untranslated region (UTR), C/E1, and NS5B regions of 60 sequences from 52 patients. Genotype distribution in the studied population was as follows: 54% (28/52) were genotype 5, 19% (10/52) were genotype 1, 19% (10/52) were genotype 4, and 2% (1/52) were genotype 3. Three of 52 (6%) individuals were infected with multiple genotypes based on the 5′UTR. Phylogenetic analysis of the 5′UTR was accurate in determining genotypes, while the C/E1 and NS5B coding region was able to differentiate both genotypes and subtypes, including an outlier group. Furthermore, this study observed the existence of distinct variants of HCV which were divergent from confirmed genotype 4 subtypes. For the first time in South Africa, this analysis has shown the presence of HCV subtypes 4k, 4q, and 4r, as well as evidence of intragenotypic recombinant 4l/4q within NS5B. In conclusion, while genotype 5a remains the predominant genotype in South Africa, the current study indicates the introduction of new subtypes and existence of variants of genotype 4 in South Africa. J. Med. Virol. 84:601–607, 2012. © 2011 Wiley Periodicals, Inc.
Transcription‐mediated amplification (TMA) for the assessment of viremia in hemodialysis patients with hepatitis C
Authors: Bastos Dauana O. , Perez Renata M. , Silva Ivonete Souza , Lemos Lara Barros , Simonetti José Pascoal , Medina‐Pestana José Osmar , Silva Antonio Eduardo B. , Ferraz Maria Lucia
Published: February 2012

The diagnosis of hepatitis C virus (HCV) infection in hemodialysis patients is difficult particularly due to the presence of intermittent viremia. The aims of this study were: (a) to determine the prevalence of intermittent viremia in hemodialysis patients with anti‐HCV antibodies who tested negative for HCV RNA by PCR at the first evaluation and (b) to evaluate the contribution of the transcription‐mediated amplification method (TMA) to the diagnosis of viremia in the PCR‐negative samples. One hundred and six patients with anti‐HCV antibodies and an initial negative result for HCV RNA by PCR were included. An additional sample was collected for a second HCV RNA test by PCR after a minimum interval of 3 months and a positive result characterized intermittent viremia. HCV RNA was investigated by TMA in the PCR‐negative sample of patients with intermittent viremia, and in the most recent sample from patients with PCR‐negative results in both determinations. Intermittent viremia was observed in 60/106 (57%) patients (57% men; age: 45 ± 10 years). Fifty‐one of the 60 negative samples from patients with intermittent viremia and 29/46 double‐negative patients were tested by TMA. This assay detected viremia in 20/51 (39%) samples of intermittent viremia and in 2/29 (7%) of double‐negative samples. The results showed that intermittent viremia is frequent in hemodialysis patients who tested negative for HCV RNA by PCR. Therefore, a second HCV RNA test is necessary for all HCV RNA‐negative patients. The TMA assay appears to be the best first screening test for viremia in this population. J. Med. Virol. 84:596–600, 2012. © 2011 Wiley Periodicals, Inc.
Characterization of glycan binding specificities of influenza B viruses with correlation with hemagglutinin genotypes and clinical features
Authors: Wang Ya‐Fang , Chang Chuan‐Fa , Chi Chia‐Yu , Wang Hsuan‐Chen , Wang Jen‐Ren , Su Ih‐Jen
Published: February 2012

The carbohydrate binding specificities are different among avian and human influenza A viruses and may affect the tissue tropism and transmission of these viruses. The glycan binding biology for influenza B, however, has not been systematically characterized. Glycan binding specificities of influenza B viral isolates were analyzed and correlated to hemagglutinin (HA) genotypes and clinical manifestations. A newly developed solution glycan array was applied to characterize the receptor binding specificities of influenza B virus clinical isolates from 2001 to 2007 in Taiwan. Thirty oligosaccharides which include α‐2,3 and α‐2,6 linkage glycans were subjected to analysis. The glycan binding patterns of 53 influenza B isolates could be categorized into three groups and were well correlated to their HA genotypes. The Yamagata‐like strains predominantly bound to α‐2,6‐linkage glycan (24:29, 83%) while Victoria‐like strains preferentially bound to both α‐2,3‐ and α‐2,6‐linkage glycans (13:24, 54%). A third group of viruses bound to sulfated glycans and these all belonged to Victoria‐like strains. Based on the HA sequences, Asn‐163, Glu‐198, Ala‐202, and Lys‐203 were conserved among Victoria‐like strains which may influence their carbohydrate recognition. The viruses bound to dual type glycans were more likely to be associated with the development of bronchopneumonia and gastrointestinal illness than those bound only to α‐2,6 sialyl glycans (P < 0.05). Glycan binding analyses provide additional information to monitor the antigenic shift, tissue tropism, and transmission capability of influenza B viruses, and will contribute to virus surveillance and vaccine strain selection. J. Med. Virol. 84:679–685, 2012. © 2011 Wiley Periodicals, Inc.
Monitoring the HTLV‐1 proviral load in the peripheral blood of asymptomatic carriers and patients with HTLV‐associated myelopathy/tropical spastic paraparesis from a Brazilian cohort: ROC curve analysis to establish the threshold for risk disease
Authors: Furtado Marina dos Santos Brito Silva , Andrade Rafaela Gomes , Romanelli Luiz Cláudio Ferreira , Ribeiro Maisa Aparecida , Ribas João Gabriel , Torres Elídio Barbosa , Barbosa‐Stancioli Edel Figueiredo , Proietti Anna Bárbara de Freitas Carneiro , Martins Marina Lobato
Published: February 2012

Human T‐lymphotropic virus 1 (HTLV‐1) infection is associated with HTLV‐associated myelopathy/tropical spastic paraparesis (HAM/TSP), which affects approximately 5% of carriers. High proviral load is a risk marker for HAM/TSP, although there is an overlap of proviral load levels in peripheral blood between asymptomatic carriers and HAM/TSP patients. In this study, receiver operating characteristic curve analysis was used to define a set point of HTLV‐1 proviral load that better indicates an increased risk for HAM/TSP. Proviral load was quantified in 75 asymptomatic carriers and 78 HAM/TSP patients in a Brazilian cohort. The cut‐off of proviral load was defined as 114 copies/104 cells, with 78.2% sensitivity to identify true HAM/TSP patients. The mean proviral load levels were not significantly different between males and females with the same clinical status, and there was no significant correlation between proviral load and age at blood sampling, age at the onset of illness, or duration of disease. In HAM/TSP patients, proviral load was significantly higher in wheelchair‐bound patients than in individuals able to walk without support and in those with the worst spinal cord injuries. Follow‐up of HTLV‐1‐infected individuals showed that proviral load was more stable in asymptomatic carriers than in HAM/TSP patients. In a cohort study, periodically quantifying proviral load in asymptomatic carriers is necessary to identify those at risk for developing neurological disease, and it is necessary for HAM/TSP patients to monitor spinal injury and progression to walking disability. The measure of proviral load in clinical practice implicates the definition of the cut‐off of proviral load and its validation during follow‐up. J. Med. Virol. 84:664–671, 2012. © 2011 Wiley Periodicals, Inc.
Comparative analysis of hepatitis B virus genotype a molecular evolution in patients infected with HBV and in patients co‐infected with HBV and HIV
Authors: Cassino L. , Torres C. , Mbayed V. , Laufer N. , Campos R.H. , Quarleri J.
Published: February 2012

HIV infection has a significant impact on the natural progression of liver disease caused by infection with hepatitis B virus (HBV), but its role in the molecular evolution of HBV is unknown. It is difficult to study the molecular evolution of HBV longitudinally considering its genomic complexity, which implies the analysis of paired samples. This study aimed to analyze the difference in the evolutionary dynamics of HBV among patients with HIV and uninfected individuals. In this study, 17 patients infected chronically with HBV were recruited, 9 of them were co‐infected with HIV. Patients were HBe antigen‐positive and infected with HBV genotype A. Paired plasma samples were collected from each patient 3 years apart, and they were compared subsequently to each other. The HBV phylogenetic inference among isolates from patients infected with HBV and co‐infected with HBV and HIV tends to cluster separately. Likewise, when comparing the HBV evolutionary rate and genetic distances, values were higher in the former in both preC/C and S genomic regions. Intra‐host analyses of HBV isolates revealed high diversity and complexity of quasispecies among patients infected with HBV exhibiting high numbers of viral variants and genetic distance. In summary, after studying the HBV molecular evolution among isolates ascribed to genotype A at inter‐ and intra‐host levels, HBV exhibited low quasispecies complexity and diversity as well as low evolutionary rates in the presence of HIV co‐infection, suggesting that the co‐infection may have an impact on the HBV molecular evolution most likely from the weakened cellular immune response. J. Med. Virol. 84:562–569, 2012. © 2011 Wiley Periodicals, Inc.
A clustering phenomenon among HCV‐1a strains among patients coinfected with HIV from Buenos Aires, Argentina
Authors: Bolcic Federico , Laufer Natalia , Trinchero Juan , Jones Leandro R. , Quarleri Jorge
Published: February 2012

The human immunodeficiency virus (HIV) and hepatitis C virus (HCV) share the same transmission routes which lead to high coinfection rates. Among HIV‐infected individuals such rates reached 21% in Argentina, being HCV‐1a the most predominant subtype. In this work, 25 HCV subtype 1a (HCV‐1a) strains from Argentinean patients coinfected with HIV were studied based on E2 and NS5A sequences. Phylogenetic analyses indicated that 12 strains were highly related to each other, constituting a highly supported (posterior probability = 0.95) monophyletic group that we called “M.” The remaining HCV strains (group dispersed or “D”) were interspersed along the phylogenetic trees. When comparing both groups of HCV‐1a, 10 amino acid differences were located in functional domains of E2 and NS5A proteins that appeared to affect eventually the peptides binding to MHC‐I molecules thus favoring immune escape and contributing to the divergence of HCV genotypes. Bayesian coalescent analyses for HCV‐1a cluster M isolates indicated that the time to the most recent common ancestor (tMRCA) overlaps with the age estimated recently for the HIV‐BF epidemic in Argentina. Furthermore, the genomic characterization based on pol gene analysis from HIV viremic patients showed that most HIV isolates from patients coinfected with HCV‐1a cluster M were BF recombinants with identical recombination patterns. In conclusion, these results suggest the presence of an HCV‐1a monophyletic cluster with a potential HIV co‐transmission by phylogenetic analyses. J. Med. Virol. 84:570–581, 2012. © 2011 Wiley Periodicals, Inc.