bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytomegalovirus immunoevasin reveals the physiological role of “missing self” recognition in natural killer cell dependent virus control in vivo
Authors: Babić Marina , Pyzik Michal , Zafirova Biljana , Mitrović Maja , Butorac Višnja , Lanier Lewis L. , Krmpotić Astrid , Vidal Silvia M. , Jonjić Stipan
Published: November 2010

Cytomegaloviruses (CMVs) are renowned for interfering with the immune system of their hosts. To sidestep antigen presentation and destruction by CD8+ T cells, these viruses reduce expression of major histocompatibility complex class I (MHC I) molecules. However, this process sensitizes the virus-infected cells to natural killer (NK) cell–mediated killing via the “missing self” axis. Mouse cytomegalovirus (MCMV) uses m152 and m06 encoded proteins to inhibit surface expression of MHC I molecules. In addition, it encodes another protein, m04, which forms complexes with MHC I and escorts them to the cell surface. This mechanism is believed to prevent NK cell activation and killing by restoring the “self” signature and allowing the engagement of inhibitory Ly49 receptors on NK cells. Here we show that MCMV lacking m04 was attenuated in an NK cell– and MHC I–dependent manner. NK cell–mediated control of the infection was dependent on the presence of NK cell subsets expressing different inhibitory Ly49 receptors. In addition to providing evidence for immunoevasion strategies used by CMVs to avoid NK cell control via the missing-self pathway, our study is the first to demonstrate that missing self–dependent NK cell activation is biologically relevant in the protection against viral infection in vivo.
Targeting of B and T lymphocyte associated (BTLA) prevents graft-versus-host disease without global immunosuppression
Authors: Albring Jörn C. , Sandau Michelle M. , Rapaport Aaron S. , Edelson Brian T. , Satpathy Ansuman , Mashayekhi Mona , Lathrop Stephanie K. , Hsieh Chyi-Song , Stelljes Matthias , Colonna Marco , Murphy Theresa L. , Murphy Kenneth M.
Published: November 2010

Graft-versus-host disease (GVHD) causes significant morbidity and mortality in allogeneic hematopoietic stem cell transplantation (aHSCT), preventing its broader application to non–life-threatening diseases. We show that a single administration of a nondepleting monoclonal antibody specific for the coinhibitory immunoglobulin receptor, B and T lymphocyte associated (BTLA), permanently prevented GVHD when administered at the time of aHSCT. Once GVHD was established, anti-BTLA treatment was unable to reverse disease, suggesting that its mechanism occurs early after aHSCT. Anti-BTLA treatment prevented GVHD independently of its ligand, the costimulatory tumor necrosis factor receptor herpesvirus entry mediator (HVEM), and required BTLA expression by donor-derived T cells. Furthermore, anti-BTLA treatment led to the relative inhibition of CD4+ forkhead box P3− (Foxp3−) effector T cell (T eff cell) expansion compared with precommitted naturally occurring donor-derived CD4+ Foxp3+ regulatory T cell (T reg cell) and allowed for graft-versus-tumor (GVT) effects as well as robust responses to pathogens. These results suggest that BTLA agonism rebalances T cell expansion in lymphopenic hosts after aHSCT, thereby preventing GVHD without global immunosuppression. Thus, targeting BTLA with a monoclonal antibody at the initiation of aHSCT therapy might reduce limitations imposed by histocompatibility and allow broader application to treatment of non–life-threatening diseases.
Relative motion of transmembrane segments S0 and S4 during voltage sensor activation in the human BKCa channel
Authors: Pantazis Antonios , Kohanteb Azadeh P. , Olcese Riccardo
Published: November 2010

Large-conductance voltage- and Ca2+-activated K+ (BKCa) channel α subunits possess a unique transmembrane helix referred to as S0 at their N terminus, which is absent in other members of the voltage-gated channel superfamily. Recently, S0 was found to pack close to transmembrane segments S3 and S4, which are important components of the BKCa voltage-sensing apparatus. To assess the role of S0 in voltage sensitivity, we optically tracked protein conformational rearrangements from its extracellular flank by site-specific labeling with an environment-sensitive fluorophore, tetramethylrhodamine maleimide (TMRM). The structural transitions resolved from the S0 region exhibited voltage dependence similar to that of charge-bearing transmembrane domains S2 and S4. The molecular determinant of the fluorescence changes was identified in W203 at the extracellular tip of S4: at hyperpolarized potential, W203 quenches the fluorescence of TMRM labeling positions at the N-terminal flank of S0. We provide evidence that upon depolarization, W203 (in S4) moves away from the extracellular region of S0, lifting its quenching effect on TMRM fluorescence. We suggest that S0 acts as a pivot component against which the voltage-sensitive S4 moves upon depolarization to facilitate channel activation.
Unitary Ca2+ current through recombinant type 3 InsP3 receptor channels under physiological ionic conditions
Authors: Vais Horia , Foskett J. Kevin , Daniel Mak Don-On
Published: November 2010

The ubiquitous inositol 1,4,5-trisphosphate (InsP3) receptor (InsP3R) channel, localized primarily in the endoplasmic reticulum (ER) membrane, releases Ca2+ into the cytoplasm upon binding InsP3, generating and modulating intracellular Ca2+ signals that regulate numerous physiological processes. Together with the number of channels activated and the open probability of the active channels, the size of the unitary Ca2+ current (iCa) passing through an open InsP3R channel determines the amount of Ca2+ released from the ER store, and thus the amplitude and the spatial and temporal nature of Ca2+ signals generated in response to extracellular stimuli. Despite its significance, iCa for InsP3R channels in physiological ionic conditions has not been directly measured. Here, we report the first measurement of iCa through an InsP3R channel in its native membrane environment under physiological ionic conditions. Nuclear patch clamp electrophysiology with rapid perfusion solution exchanges was used to study the conductance properties of recombinant homotetrameric rat type 3 InsP3R channels. Within physiological ranges of free Ca2+ concentrations in the ER lumen ([Ca2+]ER), free cytoplasmic [Ca2+] ([Ca2+]i), and symmetric free [Mg2+] ([Mg2+]f), the iCa–[Ca2+]ER relation was linear, with no detectable dependence on [Mg2+]f. iCa was 0.15 ± 0.01 pA for a filled ER store with 500 µM [Ca2+]ER. The iCa–[Ca2+]ER relation suggests that Ca2+ released by an InsP3R channel raises [Ca2+]i near the open channel to ∼13–70 µM, depending on [Ca2+]ER. These measurements have implications for the activities of nearby InsP3-liganded InsP3R channels, and they confirm that Ca2+ released by an open InsP3R channel is sufficient to activate neighboring channels at appropriate distances away, promoting Ca2+-induced Ca2+ release.
On the mechanism of CFTR inhibition by a thiazolidinone derivative
Authors: Kopeikin Zoia , Sohma Yoshiro , Li Min , Hwang Tzyh-Chang
Published: November 2010

The effects of a thiazolidinone derivative, 3-[(3-trifluoromethyl)phenyl]-5-[(4-carboxyphenyl)methylene]-2-thioxo-4-thiazolidinone (or CFTRinh-172), on cystic fibrosis transmembrane conductance regulator (CFTR) gating were studied in excised inside-out membrane patches from Chinese hamster ovary cells transiently expressing wild-type and mutant CFTR. We found that the application of CFTRinh-172 results in an increase of the mean closed time and a decrease of the mean open time of the channel. A hyperbolic relationship between the closing rate and [CFTRinh-172] suggests that CFTRinh-172 does not act as a simple pore blocker. Interestingly, the potency of inhibition increases as the open time of the channel is increased with an IC50 in the low nanomolar range for CFTR channels locked in an open state for tens of seconds. Our studies also provide evidence that CFTRinh-172 can bind to both the open state and the closed state. However, at least one additional step, presumably reflecting inhibitor-induced conformational changes, is required to shut down the conductance after the binding of the inhibitor to the channel. Using the hydrolysis-deficient mutant E1371S as a tool as the closing rate of this mutant is dramatically decreased, we found that CFTRinh-172–dependent inhibition of CFTR channel gating, in two aspects, mimics the inactivation of voltage-dependent cation channels. First, similar to the recovery from inactivation in voltage-gated channels, once CFTR is inhibited by CFTRinh-172, reopening of the channel can be seen upon removal of the inhibitor in the absence of adenosine triphosphate (ATP). Second, ATP induced a biphasic current response on inhibitor-bound closed channels as if the ATP-opened channels “inactivate” despite a continuous presence of ATP. A simplified six-state kinetic scheme can well describe our data, at least qualitatively. Several possible structural mechanisms for the effects of CFTRinh-172 will be discussed.
FILTREST3D: discrimination of structural models using restraints from experimental data
Authors: Gajda Michal J. , Tuszynska Irina , Kaczor Marta , Bakulina Anastasia Yu. , Bujnicki Janusz M.
Published: November 2010

Summary: Automatic methods for macromolecular structure prediction (fold recognition, de novo folding and docking programs) produce large sets of alternative models. These large model sets often include many native-like structures, which are often scored as false positives. Such native-like models can be more easily identified based on data from experimental analyses used as structural restraints (e.g. identification of nearby residues by cross-linking, chemical modification, site-directed mutagenesis, deuterium exchange coupled with mass spectrometry, etc.). We present a simple server for scoring and ranking of models according to their agreement with user-defined restraints.

Availability: FILTREST3D is freely available for users as a web server and standalone software at: http://filtrest3d.genesilico.pl/

Contact:  iamb@genesilico.pl

Supplementary information:  Supplementary data are available at Bioinformatics online.
Comorbid HIV and Myasthenia Gravis: Case Report and Review of the Literature
Authors: Knopf Lisa , Menkes Daniel L
Published: November 2010

A 27-year-old woman without known risk factors contracted HIV and was given highly active antiretroviral therapy in 2006. She subsequently developed myasthenia gravis (MG) that responded to treatment with pyridostigmine bromide and azathioprine. The medical literature, consisting primarily of case reports, indicates that MG occurs in relatively immunocompetent HIV-infected persons who generally present with mild MG symptoms. As such, we recommend a high index of suspicion for MG in HIV-infected patients presenting with fatigue and weakness, especially those receiving highly active antiretroviral therapy.
Prolonged Improvement After Rituximab: Two Cases of Resistant Muscle-Specific Receptor Tyrosine Kinase + Myasthenia Gravis
Authors: Burusnukul Prinyarat , Brennan Thomas D , Cupler Edward J
Published: November 2010

Forty percent to 50% of acetylcholine receptor antibody-seronegative patients with myasthenia gravis have muscle-specific receptor tyrosine kinase antibodies. Many muscle-specific receptor tyrosine kinase + myasthenia gravis patients remain refractory with conventional therapies. Rituximab is an anti-CD20 monoclonal antibody used in refractory B-cell disorders. Currently there is no standard dosing schedule for rituximab. We present two muscle-specific receptor tyrosine kinase + myasthenia gravis patients clinically refractory to conventional therapy who, after a single course of rituximab, became asymptomatic and discontinued all medication.
Morphology, testes development and behaviour of unusual triploid males in microchromosome‐carrying clones of Poecilia formosa
Authors: Lamatsch D. K. , Stöck M. , Fuchs R. , Döbler M. , Wacker R. , Parzefall J. , Schlupp I. , Schartl M.
Published: November 2010

In a microchromosome‐carrying laboratory stock of the normally all‐female Amazon molly Poecilia formosa triploid individuals were obtained, all of which spontaneously developed into males. A comparison of morphology of the external and internal insemination apparatus and the gonads, sperm ploidy and behaviour, to laboratory‐bred F1 hybrids revealed that the triploid P. formosa males, though producing mostly aneuploid sperm, are partly functional males that differ mainly in sperm maturation and sexual motivation from gonochoristic P. formosa males.
Cod Gadus morhua and climate change: processes, productivity and prediction
Authors: Brander K. M.
Published: November 2010

Environmental factors act on individual fishes directly and indirectly. The direct effects on rates and behaviour can be studied experimentally and in the field, particularly with the advent of ever smarter tags for tracking fishes and their environment. Indirect effects due to changes in food, predators, parasites and diseases are much more difficult to estimate and predict. Climate can affect all life‐history stages through direct and indirect processes and although the consequences in terms of growth, survival and reproductive output can be monitored, it is often difficult to determine the causes. Investigation of cod Gadus morhua populations across the whole North Atlantic Ocean has shown large‐scale patterns of change in productivity due to lower individual growth and condition, caused by large‐scale climate forcing. If a population is being heavily exploited then a drop in productivity can push it into decline unless the level of fishing is reduced: the idea of a stable carrying capacity is a dangerous myth. Overexploitation can be avoided by keeping fishing mortality low and by monitoring and responding rapidly to changes in productivity. There are signs that this lesson has been learned and that G. morhua will continue to be a mainstay of the human diet.
Variation with reproductive status of PGE2 receptor immunoreactivities in the Bostrichthys sinensis olfactory system
Authors: Lai X.‐J. , Hong W.‐S.
Published: November 2010

The role of PGE2 as a putative sex pheromone in Chinese black sleeper Bostrichthys sinensis was investigated, using immunocytochemistry and how the immunoreactivities of the prostaglandin E2 (PGE2) receptor subtypes EP1, EP2, EP3 and EP4 varied with reproductive status in the olfactory system was determined. The results showed that PGE2 receptors were present in the whole of the olfactory system of B. sinensis, and that the number of receptors was linked to the reproductive status of the fish. The densities of EP1 immunoreactivity in the olfactory epithelium of mature fish were significantly (P < 0·01) higher than those in immature fish of both sexes, and the densities of EP2 and EP3 immunoreactivities in mature fish were higher (but not significantly) than those in immature fish of both sexes. In the olfactory nerve, the density of EP2 immunoreactivity in mature fish was higher (but not significantly) than that in immature fish in both sexes. In the olfactory bulb, the densities of EP1–4 immunoreactivities in mature females were significantly (P < 0·05 or <0·01) higher than those in immature females, and the density of EP4 immunoreactivity in mature males was significantly (P < 0·01) higher than that in immature males. As far as is known, the present study is the first report of the immunoreactivities of PGE2 receptor subtypes in the olfactory system of a teleost, and offers new findings regarding the role of PGE2 as sex pheromone and hormone in the reproductive behaviour and pheromonal communication of B. sinensis.
Size‐mediated response to public cues of predation risk in a tropical stream fish
Authors: Elvidge C. K. , Ramnarine I. W. , Godin J.‐G. J. , Brown G. E.
Published: November 2010

In order to investigate any size‐dependent differences between behavioural patterns, wild‐caught Hart's rivulusRivulus hartiiof varying sizes were exposed to chemical alarm cues extracted from the skin of conspecifics or heterospecificPoecilia reticulata, or a tank water control, in a series of laboratory trials. In response to conspecific alarm cues,R. hartiisubjects of the range of body sizes tested exhibited consistent, size‐independent antipredator behaviours that were characterized by decreased locomotory activity and foraging levels and increased refuging behaviour. Conversely, focalR. hartiidemonstrated significant size‐dependent trends in response to heterospecific alarm cues, with smaller individuals exhibiting antipredator responses and larger individuals shifting their behaviour to increased levels of activity consistent with a foraging, or predatory, response. These results show that the behavioural responses of individualR. hartiito publicly available chemical alarm cues from heterospecifics are mediated by the size of the receiver.
Probability and predictors of remission from life‐time nicotine, alcohol, cannabis or cocaine dependence: results from the National Epidemiologic Survey on Alcohol and Related Conditions
Authors: Lopez‐Quintero Catalina , Hasin Deborah S. , de los Cobos José Pérez , Pines Abigail , Wang Shuai , Grant Bridget F. , Blanco Carlos
Published: November 2010

Aim  To estimate the general and racial/ethnic specific cumulative probability of remission from nicotine alcohol cannabis or cocaine dependence, and to identify predictors of remission across substances.

Design  Data were collected from structured diagnostic interviews using the Alcohol Use Disorder and Associated Disabilities Interview Schedule—DSM‐IV version.

Setting  The 2001–2002 National Epidemiological Survey of Alcohol and Related Conditions (NESARC) surveyed a nationally representative sample from US adults (n = 43 093) selected in a three‐stage sampling design.

Participants  The subsamples of individuals with life‐time DSM‐IV diagnosis of dependence on nicotine (n = 6937), alcohol (n = 4781), cannabis (n = 530) and cocaine (n = 408).

Measurements  Cumulative probability estimates of dependence remission for the general population and across racial/ethnic groups. Hazard ratios for remission from dependence.

Findings  Life‐time cumulative probability estimates of dependence remission were 83.7% for nicotine, 90.6% for alcohol, 97.2% for cannabis and 99.2% for cocaine. Half of the cases of nicotine, alcohol, cannabis and cocaine dependence remitted approximately 26, 14, 6 and 5 years after dependence onset, respectively. Males, Blacks and individuals with diagnosis of personality disorders and history of substance use comorbidity exhibited lower hazards of remission for at least two substances.

Conclusions  A significant proportion of individuals with dependence on nicotine, alcohol, cannabis or cocaine achieve remission at some point in their life‐time, although the probability and time to remission varies by substance and racial/ethnic group. Several predictors of remission are shared by at least two substances, suggesting that the processes of remission overlap. The lower rates of remission of individuals with comorbid personality or substance use disorders highlight the need for providing coordinated psychiatric and substance abuse interventions.
Monoclonal antibodies to B and T lymphocyte attenuator (BTLA) have no effect on in vitro B cell proliferation and act to inhibit in vitro T cell proliferation when presented in a cis, but not trans, format relative to the activating stimulus
Authors: Zhang M , Howard K , Winters A , Steavenson S , Anderson S , Smelt S , Doellgast G , Sheelo C , Stevens J , Kim H , Hamburger A , Sein A , Caughey D J , Lee F , Hsu H , Siu G , Byrne F R
Published: November 2010

B and T lymphocyte attenuator (BTLA) is an immunoglobulin superfamily member surface protein expressed on B and T cells. Its ligand, herpesvirus entry mediator (HVEM), is believed to act as a monomeric agonist that signals via the CRD1 of HVEM to inhibit lymphocyte activation: HVEM is also the receptor for lymphotoxin-α and LIGHT, which both bind in the CRD2 and CRD3 domains of the HVEM molecule, and for CD160 which competes with BTLA. We have shown that recombinant HVEM and a panel of different monoclonal antibodies specifically bind murine BTLA on both B and T cells and that some antibodies inhibit anti-CD3ε-induced T cell proliferation in vitro, but only when constrained appropriately with a putatively cross-linking reagent. The antibodies had no significant effect on in vitro T cell proliferation in a mixed lymphocyte reaction (MLR) assay nor on in vitro DO11.10 antigen-induced T cell proliferation. None of these antibodies, nor HVEM-Fc, had any significant effect on in vitro B cell proliferation induced by anti-immunoglobulin M antibodies (±anti-CD40) or lipopolysaccharide. We further elucidated the requirements for inhibition of in vitro T cell proliferation using a beads-based system to demonstrate that the antibodies that inhibited T cell proliferation in vitro were required to be presented to the T cell in a cis, and not trans, format relative to the anti-CD3ε stimulus. We also found that antibodies that inhibited T cell proliferation in vitro had no significant effect on the antibody captured interleukin-2 associated with the in vivo activation of DO11.10 T cells transferred to syngeneic recipient BALB/c mice. These data suggest that there may be specific structural requirements for the BTLA molecule to exert its effect on lymphocyte activation and proliferation.
The mode of actions of the Adacolumn therapeutic leucocytapheresis in patients with inflammatory bowel disease: a concise review
Authors: Hanai H , Takeda Y , Eberhardson M , Gruber R , Saniabadi A R , Winqvist O , Lofberg R
Published: November 2010

Patients with active inflammatory bowel disease (IBD) have elevated and activated myeloid leucocytes which infiltrate the colonic mucosa in vast numbers. Myeloid leucocytes such as the CD14+CD16+ monocytes are major sources of tumour necrosis factor (TNF)-α, and therefore selective granulocyte/monocyte (GM) adsorption (GMA) should promote remission or enhance efficacy of pharmacological therapy. However, studies in IBD have reported both impressive as well as disappointing efficacy outcomes, indicating that patients' demographic factors might determine responders or non-responders to GMA. Nonetheless, this non-drug intervention has an excellent safety profile, and therapeutic GMA is expected to expand. In this review, attempts have been made to compile an update on the mode of actions (MoA) of the Adacolumn GMA. The MoA of GMA appears to be more than adsorption of excess neutrophils and TNF-producing CD14+CD16+ monocytes per se. Adsorbed GMs release interleukin (IL)-1 receptor antagonist, hepatocyte growth factor and soluble TNF receptors, which are anti-inflammatory. Additionally, a sustained increase in lymphocytes including the regulatory CD4+CD25+ T cells (lymphocyte sparing) is seen post-GMA. The impact of GMA on the immune system is potentially very interesting in the context of treating immune-related diseases. Future studies are expected to add intriguing insights to the MoA of GMA.
Effect of sugarcane harvesting systems on soil carbon stocks in Brazil: an examination of existing data
Authors: Cerri C. C. , Galdos M. V. , Maia S. M. F. , Bernoux M. , Feigl B. J. , Powlson D. , Cerri C. E. P.
Published: November 2010

Agricultural management practices that promote net carbon (C) accumulation in the soil have been considered as an important potential mitigation option to combat global warming. The change in the sugarcane harvesting system, to one which incorporates C into the soil from crop residues, is the focus of this work. The main objective was to assess and discuss the changes in soil organic C stocks caused by the conversion of burnt to unburnt sugarcane harvesting systems in Brazil, when considering the main soils and climates associated with this crop. For this purpose, a dataset was obtained from a literature review of soils under sugarcane in Brazil. Although not necessarily from experimental studies, only paired comparisons were examined, and for each site the dominant soil type, topography and climate were similar. The results show a mean annual C accumulation rate of 1.5 Mg ha−1 year−1 for the surface to 30‐cm depth (0.73 and 2.04 Mg ha−1 year−1 for sandy and clay soils, respectively) caused by the conversion from a burnt to an unburnt sugarcane harvesting system. The findings suggest that soil should be included in future studies related to life cycle assessment and C footprint of Brazilian sugarcane ethanol.
Clinical significance and prognostic value of chromatin assembly factor‐1 overexpression in human solid tumours
Authors: Polo Sophie E , Theocharis Stamatios E , Grandin Laure , Gambotti Laetitia , Antoni Guillemette , Savignoni Alexia , Asselain Bernard , Patsouris Efstratios , Almouzni Geneviève
Published: November 2010

Polo S E, Theocharis S E, Grandin L, Gambotti L, Antoni G, Savignoni A, Asselain B, Patsouris E & Almouzni G (2010) Histopathology 57, 716–724 Clinical significance and prognostic value of chromatin assembly factor‐1 overexpression in human solid tumours

Aims:  Chromatin assembly factor‐1 (CAF‐1), whose function is critical for maintaining chromatin stability during DNA replication and repair, has been identified as a proliferation marker in breast cancer. The aim was to investigate CAF‐1 as a proliferation marker in a wide variety of solid tumours, and to assess its potential value in predicting clinical outcome.

Methods and results:  Using immunocytochemistry on paraffin‐embedded tissue sections, the CAF‐1 labelling index was compared with known proliferation markers Ki‐67 and minichromosome maintenance (MCM), and its association with clinicopathological data and patients’ outcome analysed. CAF‐1 expression showed a strong positive correlation with Ki‐67, used routinely to detect proliferating cells, while it generally displayed weaker correlations with MCM markers, known to label cells with replicative potential. CAF‐1 expression was associated significantly with histological grade in breast, cervical, endometrial and renal cell carcinomas, and with disease stage in endometrial and renal carcinomas. Furthermore, high expression of CAF‐1 was an independent predictor of adverse clinical outcome in renal, endometrial and cervical carcinomas.

Conclusions:  CAF‐1 is a proliferation marker in various malignant tumours with prognostic value in renal, endometrial and cervical carcinomas, which supports the value of CAF‐1 as a clinical marker of cancer progression.
Immunohistochemical detection of arginine methylated proteins (MeRP) in archival tissues
Authors: Vezzalini Marzia , Aletta John M , Beghelli Stefania , Moratti Elisabetta , Della Peruta Marco , Mafficini Andrea , Mojica Wilfrido D , Mombello Aldo , Scarpa Aldo , Sorio Claudio
Published: November 2010

Vezzalini M, Aletta J M, Beghelli S, Moratti E, Della Peruta M, Mafficini A, Mojica W D, Mombello A, Scarpa A & Sorio C (2010) Histopathology57, 725–733  Immunohistochemical detection of arginine methylated proteins (MeRP) in archival tissues

Aims:  To (i) determine whether methylarginine‐specific antibodies can be employed for standard immunohistochemical analysis of paraffin‐embedded tissues, (ii) analyse methylarginine expression in normal and neoplastic tissues and (iii) correlate methylarginine expression with that of protein arginine methyltransferase (PRMT1), the predominant cellular arginine methyltransferase.

Methods and results:  Immunohistochemistry of normal and cancer tissues was performed utilizing three commercial polyclonal antibodies: anti‐methylarginine‐specific antibody (anti‐mRG) raised against a methylarginine peptide, Control antibody (anti‐RG), a control antiserum raised against a corresponding arginine peptide without any methylated residues and anti‐PRMT1. Nuclear and/or cytoplasmic methylarginine expression was detected in all keratinized and non‐keratinized epithelia. A preliminary survey of a series of thyroid, pancreatic, colonic and gastric cancers identified a different pattern of methylarginine expression in comparison with normal tissue. A correlation between methylarginine staining and PRMT1 expression was found in all normal and cancer tissues analysed.

Conclusion:  Methylarginine‐specific antibodies are capable of recognizing methylarginine proteins (MeRP) in paraffin‐embedded tissues. Methylarginine proteins are expressed widely and show differences in subcellular localization in various organs and neoplastic conditions. The efficient detection of methylproteins by standard immunohistochemistry provides a new tool to investigate the role of methylarginine proteins (MeRP) in biological processes including carcinogenesis.
Deregulation of miR‐519a, 153, and 485‐5p and its clinicopathological relevance in ovarian epithelial tumours
Authors: Kim Tae Heon , Kim Yu Kyung , Kwon Youngdo , Heo Jin Hyung , Kang Haeyoun , Kim Gwangil , An Hee Jung
Published: November 2010

Kim T H, Kim Y K, Kwon Y, Heo J H, Kang H, Kim G & An H J (2010) Histopathology57, 734–743  Deregulation of miR‐519a, 153, and 485‐5p and its clinicopathological relevance in ovarian epithelial tumours

Aims:  The pathological and clinical significance of aberrant miRNA expression in ovarian tumours has yet to be adequately documented. The aim of this study was to assess the differences in miRNA expression of human ovarian tumours according to histological subtype, and to determine whether miRNAs are potential diagnostic and prognostic markers in ovarian cancers.

Method and results:  The miRNA expression profiles of 103 human ovarian tumours were evaluated. Via a bead‐based miRNA microarray, five aberrant miRNAs were selected which were expressed differentially in malignant serous tumours from borderline and benign ovarian tumours, including miRNA (miR)‐519a, miR‐18b (up‐regulation) and miR‐153, miR‐511 and miR‐485‐5p (down‐regulation). We conducted quantitative real‐time reverse transcription–polymerase chain reaction (qRT–PCR) in order to confirm that these miRNAs are differentially expressed in different histological subtypes of ovarian tumours, and compared the expression profiles of these miRNAs between different clinical subsets. The expression of these miRNAs was correlated with clinicopathological parameters. miR‐519a, miR‐153 and miR‐485‐5p were differentially expressed in four major histotypes of ovarian cancers (P < 0.05), which suggests that they might be of potential importance as diagnostic biomarkers. Down‐regulation of miR‐153 and miR‐485‐5p was correlated significantly with FIGO grade 3 (P < 0.05). Down‐regulation of miR‐153 and up‐regulation of miR‐519a were correlated significantly with advanced clinical stage (P < 0.05). The results of Kaplan–Meier survival analysis indicated that the higher expression of miR‐519a in late stage serous carcinoma was associated significantly with poor progression‐free survival (P = 0.0058).

Conclusions:  A significant correlation was detected between the deregulation of specific types of miRNAs, such as miR‐519a, miR‐153 and miR‐485‐5p, and clinical variables as well as histological subtypes in ovarian cancers. Hence, these miRNAs may perform functions as diagnostic or prognostic biomarkers.
Therapeutic molecular targets in human chondrosarcoma
Authors: Jamil Nuor , Howie Sarah , Salter Donald M.
Published: November 2010

Chondrosarcomas are malignant cartilage tumours. They are poorly responsive to chemotherapy and radiotherapy. Treatment is usually limited to surgical resection; however, survival of patients with high‐grade chondrosarcoma is poor, even with wide surgical resection. Induction of apoptosis in chondrosarcoma cells, either directly or by enhancement of the response to chemotherapeutic drugs and radiation, may be a route by which outcome can be improved. In this article, we review potential molecular targets that regulate chondrocyte apoptosis and discuss the experimental evidence for their utility.
The fertility restorer gene, Rf2, for Lead Rice‐type cytoplasmic male sterility of rice encodes a mitochondrial glycine‐rich protein
Authors: Itabashi Etsuko , Iwata Natsuko , Fujii Sota , Kazama Tomohiko , Toriyama Kinya
Published: November 2010

Cytoplasmic male sterility (CMS) is associated with a mitochondrial mutation that causes an inability to produce fertile pollen. The fertility of CMS plants is restored in the presence of a nuclear‐encoded fertility restorer (Rf) gene. In Lead Rice‐type CMS, discovered in the indica variety ‘Lead Rice’, fertility of the CMS plant is restored by the single nuclear‐encoded gene Rf2 in a gametophytic manner. We performed map‐based cloning of Rf2, and proved that it encodes a protein consisting of 152 amino acids with a glycine‐rich domain. Expression of Rf2 mRNA was detected in developing and mature anthers. An RF2–GFP fusion was shown to be targeted to mitochondria. Replacement of isoleucine by threonine at amino acid 78 of the RF2 protein was considered to be the cause of functional loss in the rf2 allele. As Rf2 does not encode a pentatricopeptide repeat protein, unlike a majority of previously identified Rf genes, the data from this study provide new insights into the mechanism for restoring fertility in CMS.
Polycomb proteins regulate the quantitative induction of VERNALIZATION INSENSITIVE 3 in response to low temperatures
Authors: Jean Finnegan E. , Bond Donna M. , Buzas Diana M. , Goodrich Justin , Helliwell Christopher A. , Tamada Yosuke , Yun Jae‐Young , Amasino Richard M. , Dennis Elizabeth S.
Published: November 2010

Vernalization, the promotion of flowering in response to low temperatures, is one of the best characterized examples of epigenetic regulation in plants. The promotion of flowering is proportional to the duration of the cold period, but the mechanism by which plants measure time at low temperatures has been a long‐standing mystery. We show that the quantitative induction of the first gene in the Arabidopsis vernalization pathway, VERNALIZATION INSENSITIVE 3 (VIN3), is regulated by the components of Polycomb Response Complex 2, which trimethylates histone H3 lysine 27 (H3K27me3). In differentiated animal cells, H3K27me3 is mostly associated with long‐term gene repression, whereas, in pluripotent embyonic stem cells, many cell lineage‐specific genes are inactive but exist in bivalent chromatin that carries both active (H3K4me3) and repressive (H3K27me3) marks on the same molecule. During differentiation, bivalent domains are generally resolved to an active or silent state. We found that H3K27me3 maintains VIN3 in a repressed state prior to cold exposure; this mark is not removed during VIN3 induction. Instead, active VIN3 is associated with bivalently marked chromatin. The continued presence of H3K27me3 ensures that induction of VIN3 is proportional to the duration of the cold, and that plants require prolonged cold to promote the transition to flowering. The observation that Polycomb proteins control VIN3 activity defines a new role for Polycomb proteins in regulating the rate of gene induction.
Susceptibility of community‐acquired pathogens to antibiotics in Africa and Asia in neonates – an alarmingly short review
Authors: Lubell Yoel , Ashley Elizabeth A. , Turner Claudia , Turner Paul , White Nicholas J.
Published: November 2010

Objective To assess the susceptibility of community‐acquired pathogens in neonatal sepsis to commonly prescribed antibiotics in sub‐Saharan Africa and Asia since 2002.

Methods Literature review in PubMed and Embase. Susceptibility was estimated for pathogens individually and stratified by region. Isolates were also classified into Gram positive and Gram negative pathogens to estimate their pooled susceptibility.

Results and conclusions Only nine studies met the inclusion criteria. The available data indicated poor susceptibility to almost all commonly used antibiotics in pathogens such asStaphylococcus aureusandKlebsiellaspp. OnlyStreptococcus pneumoniaeexhibited good susceptibility to all drugs other than cotrimoxazole. The extreme scarcity of data prevents drawing any firm conclusions beyond the urgent need for more studies to identify the best treatments for neonatal sepsis in the developing world.
Patients’ level of knowledge measured five days after kidney transplantation
Authors: Urstad Kristin H. , Andersen Marit H. , Øyen Ole , Moum Torbjørn , Wahl Astrid K.
Published: November 2010

Urstad KH, Andersen MH, Øyen O, Moum T, Wahl AK. Patients’ level of knowledge measured five days after kidney transplantation. Clin Transplant 2011: 25: 646–652. © 2010 John Wiley & Sons A/S.

Abstract:  Background:  Kidney recipients’ knowledge is important in terms of coping with short‐term problems posed by transplantation and the long‐term outcome. Little attention has been given to the development of instruments for measuring patient’s knowledge in this field.

Aim:  The purpose of this study was to describe the development of a knowledge questionnaire for kidney recipients and to explore possible factors related to the knowledge level.

Methods:  The sample consisted of 159 kidney recipients at a Norwegian transplant center, answering the questionnaire five d post‐transplantation.

Results:  The questionnaire was generated on the basis of literature review and clinical experience – and was pilot tested. Mean score of the questionnaire was 11, of 19 obtainable points. Longer duration of kidney disease was significantly correlated with an increased knowledge level, whereas the longer the time on dialysis prior to transplantation and post‐operative complications were found to have significantly negative impact on total knowledge score.

Conclusions:  The positive impact of disease duration may suggest that insight gained over time makes patients better prepared for the transplantation itself and for life post‐transplant. However, the negative impact of dialysis duration could be attributed to impaired cognitive function imposed by chronic dialysis treatment. Further research is needed regarding the questionnaire’s responsiveness to educational interventions.
Clinical features of late onset non‐infectious pulmonary complications following pediatric allogeneic hematopoietic stem cell transplantation
Authors: Park Meerim , Koh Kyung Nam , Kim Bo Eun , Im Ho Joon , Seo Jong Jin
Published: November 2010

Park M, Koh KN, Kim BE, Im HJ, Seo JJ. Clinical features of late onset non‐infectious pulmonary complications following pediatric allogeneic hematopoietic stem cell transplantation.  Clin Transplant 2011: 25: E168–E176. © 2010 John Wiley & Sons A/S.

Abstract:  Background:  Late onset non‐infectious pulmonary complications (LONIPCs) are major causes of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo‐HSCT). We evaluated the incidence and the outcomes of LONIPCs in children who underwent allo‐HSCT.

Methods:  Data for 143 children who underwent allo‐HSCT at Asan Medical Center between April 2002 and April 2009 were reviewed, and the 127 who survived more than three months were enrolled.

Results:  Eleven (8.7%) developed LONIPCs at a median eight months (range 3–14 months) after allo‐HSCT, presenting with cough and dyspnea. Six had bronchiolitis obliterans and five had idiopathic pneumonia syndrome. FVC declined more significantly in LONIPC than in non‐LONIPC patients three months after HSCT. A significant risk factor for the development of LONIPCs was chronic graft‐versus‐host disease (GVHD) (p = 0.002). At a median follow‐up of 36 months, the three‐yr overall survival rate in LONIPC patients was significantly lower than that of non‐LONIPC patients (37.4% vs. 72.7%, p = 0.02). The major cause of death was respiratory failure.

Conclusions:  Along with chronic GVHD, deterioration of pulmonary function at three months after HSCT may help in the early identification of patients at risk of subsequent LONIPCs. Considering the poor prognosis of LONIPC, strategies should be aimed at their prevention.
Glucose‐6‐phosphate dehydrogenase‐dependent hydrogen peroxide production is involved in the regulation of plasma membrane H+‐ATPase and Na+/H+ antiporter protein in salt‐stressed callus from Carex moorcroftii
Authors: Li Jisheng , Chen Guichen , Wang Xiaomin , Zhang Yanli , Jia Honglei , Bi Yurong
Published: November 2010

Glucose‐6‐phosphate dehydrogenase (G6PDH) is important for the activation of plant resistance to environmental stresses, and ion homeostasis is the physiological foundation for living cells. In this study, we investigated G6PDH roles in modulating ion homeostasis under salt stress in Carex moorcroftii callus. G6PDH activity increased to its maximum in 100 mM NaCl treatment and decreased with further increased NaCl concentrations. K+/Na+ ratio in 100 mM NaCl treatment did not exhibit significant difference compared with the control; however, in 300 mM NaCl treatment, it decreased. Low‐concentration NaCl (100 mM) stimulated plasma membrane (PM) H+‐ATPase and NADPH oxidase activities as well as Na+/H+ antiporter protein expression, whereas high‐concentration NaCl (300 mM) decreased their activity and expression. When G6PDH activity and expression were reduced by glycerol treatments, PM H+‐ATPase and NADPH oxidase activities, Na+/H+ antiporter protein level and K+/Na+ ratio dramatically decreased. Simultaneously, NaCl‐induced hydrogen peroxide (H2O2) accumulation was abolished. Exogenous application of H2O2 increased G6PDH, PM H+‐ATPase and NADPH oxidase activities, Na+/H+ antiporter protein expression and K+/Na+ ratio in the control and glycerol treatments. Diphenylene iodonium (DPI), the NADPH oxidase inhibitor, which counteracted NaCl‐induced H2O2 accumulation, decreased G6PDH, PM H+‐ATPase and NADPH oxidase activities, Na+/H+ antiporter protein level and K+/Na+ ratio. Western blot result showed that G6PDH expression was stimulated by NaCl and H2O2, and blocked by DPI. Taken together, G6PDH is involved in H2O2 accumulation under salt stress. H2O2, as a signal, upregulated PM H+‐ATPase activity and Na+/H+ antiporter protein level, which subsequently resulted in the enhanced K+/Na+ ratio. G6PDH played a central role in the process.
Clinical evaluation of chronic nephrotoxicity of long‐term cyclosporine A treatment in adult patients with steroid‐dependent nephrotic syndrome
Authors: OKADA TOMONARI , MATSUMOTO HIROSHI , NAGAOKA YUME , TOMARU RYO , IWASAWA HIDEAKI , WADA TOSHIKAZU , NAKAO TOSHIYUKI
Published: November 2010

Aim:  Chronic nephrotoxicity of long‐term cyclosporine A (CsA) treatment is a matter of concern in patients with steroid‐dependent nephrotic syndrome (SDNS).

Methods:  Twenty‐eight adult NS patients (25, minimal‐change nephrotic syndrome (NS); three, focal‐segmental glomerulosclerosis) were divided into three groups. Group A was continuously treated with CsA for more than 5 years (143 ± 40 months, 1.3 ± 0.4 mg/kg per day at final analysis, n = 12); group B had been previously treated with CsA (70 ± 27 months, n = 6); and group C had been treated with corticosteroids alone (n = 10). The clinical variables related to chronic CsA nephrotoxicity were examined.

Results:  In groups A and B, estimated glomerular filtration rate decreased from 86 ± 22 and 107 ± 17 to 83 ± 23 and 88 ± 13 mL/min per 1.73 m2, respectively, at final analysis (both P < 0.05). Serum magnesium levels in group A were significantly lower than those in group B or C (A, 1.78 ± 0.16 mg/dL; B, 2.00 ± 0.14 mg/dL; C, 2.03 ± 0.10 mg/dL; A vs B, C, P < 0.01), and a significant correlation between these and the duration of CsA treatment was found (r = −0.68, P < 0.001). There was a trend towards a correlation between the duration of CsA administration and urinary α1‐microglobulin (r = 0.38, P = 0.07).

Conclusion:  Mild decrease in renal function and hypomagnesemia were found in adult SDNS patients with long‐term CsA treatment. Careful monitoring of renal function, blood pressure and serum magnesium levels is necessary.
The next generation: Using new sequencing technologies to analyse gene regulation
Authors: CULLUM Rebecca , ALDER Olivia , HOODLESS Pamela A.
Published: November 2010

Next generation sequencing (NGS) has pushed back the limitations of prior sequencing technologies to advance genomic knowledge infinitely by allowing cost‐effective, rapid sequencing to become a reality. Genome‐wide transcriptional profiling can be achieved using NGS with either Tag‐Seq, in which short tags of cDNA represent a gene, or RNA‐Seq, in which the entire transcriptome is sequenced. Furthermore, the level and diversity of miRNA within different tissues or cell types can be monitored by specifically sequencing small RNA. The biological mechanisms underlying differential gene regulation can also be explored by coupling chromatin immunoprecipitation with NGS (ChIP‐Seq). Using this methodology genome‐wide binding sites for transcription factors, RNAP II, epigenetic modifiers and the distribution of modified histones can be assessed. The superior, high‐resolution data generated by adopting this sequencing technology allows researchers to distinguish the precise genomic location bound by a protein and correlate this with observed gene expression patterns. Additional methods have also been established to examine other factors influencing gene regulation such as DNA methylation or chromatin conformation on a genome‐wide scale. Within any research setting, these techniques can provide relevant data and answer numerous questions about gene expression and regulation. The advances made by pairing NGS with strategic experimental protocols will continue to impact the research community.
Medical visits of childhood cancer survivors in Japan: A cross‐sectional survey
Authors: Ishida Yasushi , Ozono Shuichi , Maeda Naoko , Okamura Jun , Asami Keiko , Iwai Tsuyako , Kamibeppu Kiyoko , Sakamoto Naoko , Kakee Naoko , Horibe Keizo
Published: November 2010

Background:  Although more children with cancer continue to be cured, these survivors experience various late effects. Details of the medical visit behaviors of childhood cancer survivors (CCS) in adulthood remain to be elucidated.

Methods:  In order to examine medical visits in the past and future of CCS, we performed a cross‐sectional survey with self‐rating questionnaires on medical visits of CCS compared with control groups (their siblings and the general population).

Results:  Questionnaires were completed by 185 CCS, 72 of their siblings and 1000 subjects from the general population and the results were analyzed. Mean ages at this survey and the duration after therapy completions of CCS were 23 and 12 years, respectively. We found that the previous treatment hospitals (where CCS were treated for their cancer) were the most commonly visited medical facilities for the CCS group (74% for female patients and 64% for male patients) and more than half of the CCS preferred to continue visiting the previous treatment hospital with enough satisfaction in Japan. The multivariate analysis showed that female sex and relapse were significantly associated with the past visits to the previous treatment hospital and that the CCS with brain tumors or bone/soft tissue sarcomas and CCS with any late effects tended to continue the relationships with the hospital. In addition female sex was also significantly associated with desired future visits to the previous treatment hospital. On the other hand, the married CCS tended to be disinclined to visit the hospital it in the future.

Conclusions:  In order to optimize risk‐based care and promote health for CCS after adulthood, we should discuss the medical transition with CCS and their parents.