bluuebunny/crossref_metadata_2025 huggingface dataset:
The Prp8 RNase H-like domain inhibits Brr2-mediated U4/U6 snRNA unwinding by blocking Brr2 loading onto the U4 snRNA
Authors: Mozaffari-Jovin Sina , Santos Karine F. , Hsiao He-Hsuan , Will Cindy L. , Urlaub Henning , Wahl Markus C. , Lührmann Reinhard
Published: November 2012

The spliceosomal RNA helicase Brr2 catalyzes unwinding of the U4/U6 snRNA duplex, an essential step for spliceosome catalytic activation. Brr2 is regulated in part by the spliceosomal Prp8 protein by an unknown mechanism. We demonstrate that the RNase H (RH) domain of yeast Prp8 binds U4/U6 small nuclear RNA (snRNA) with the single-stranded regions of U4 and U6 preceding U4/U6 stem I, contributing to its binding. Via cross-linking coupled with mass spectrometry, we identify RH domain residues that contact the U4/U6 snRNA. We further demonstrate that the same single-stranded region of U4 preceding U4/U6 stem I is recognized by Brr2, indicating that it translocates along U4 and first unwinds stem I of the U4/U6 duplex. Finally, we show that the RH domain of Prp8 interferes with U4/U6 unwinding by blocking Brr2's interaction with the U4 snRNA. Our data reveal a novel mechanism whereby Prp8 negatively regulates Brr2 and potentially prevents premature U4/U6 unwinding during splicing. They also support the idea that the RH domain acts as a platform for the exchange of U6 snRNA for U1 at the 5′ splice site. Our results provide insights into the mechanism whereby Brr2 unwinds U4/U6 and show how this activity is potentially regulated prior to spliceosome activation.
Mechanism for epigenetic variegation of gene expression at yeast telomeric heterochromatin
Authors: Kitada Tasuku , Kuryan Benjamin G. , Tran Nancy Nga Huynh , Song Chunying , Xue Yong , Carey Michael , Grunstein Michael
Published: November 2012

Yeast contains heterochromatin at telomeres and the silent mating-type loci (HML/HMR). Genes positioned within the telomeric heterochromatin of Saccharomyces cerevisiae switch stochastically between epigenetically bistable ON and OFF expression states. Important aspects of the mechanism of variegated gene expression, including the chromatin structure of the natural ON state and the mechanism by which it is maintained, are unknown. To address this issue, we developed approaches to select cells in the ON and OFF states. We found by chromatin immunoprecipitation (ChIP) that natural ON telomeres are associated with Rap1 binding and, surprisingly, also contain known characteristics of OFF telomeres, including significant amounts of Sir3 and H4K16 deacetylated nucleosomes. Moreover, we found that H3K79 methylation (H3K79me), H3K4me, and H3K36me, which are depleted from OFF telomeres, are enriched at ON telomeres. We demonstrate in vitro that H3K79me, but not H3K4me or H3K36me, disrupts transcriptional silencing. Importantly, H3K79me does not significantly reduce Sir complex binding in vivo or in vitro. Finally, we show that maintenance of H3K79me at ON telomeres is dependent on transcription. Therefore, although Sir proteins are required for silencing, we propose that epigenetic variegation of telomeric gene expression is due to the bistable enrichment/depletion of H3K79me and not the fluctuation in the amount of Sir protein binding to nucleosomes.
Two-Photon Sodium Imaging in Dendritic Spines
Authors: Rose Christine R.
Published: November 2012

Neuronal activity causes substantial postsynaptic Na+ transients. However, the physiological consequences of such Na+ transients remain largely unknown. High-resolution Na+ imaging is pivotal to study these questions, and, up to now, two-photon imaging with the fluorescent Na+ indicator sodium-binding benzofuran isophthalate (SBFI) has been the method of choice. When introduced into individual neurons in acute tissue slices, SBFI enables measurements of [Na+]i transients in dendrites and spines. This technique is also suitable for the determination of [Na+]i transients in other cell types, such as glial cells, with high spatial resolution, and is likely to be useful for measurement of Na+ signals in vivo. This protocol provides guidelines and tips for measuring dynamic Na+ concentrations within neurons, as well as an in situ calibration procedure. Because the basic properties of SBFI inside a cell differ significantly from the same properties in vitro, in situ calibrations are required to obtain meaningful experimental results.
Estimating Volume in Biological Structures
Authors: West Mark J.
Published: November 2012

Estimates of volume can be useful in comparative and experimental studies of tissues and organs. In this article, a simple procedure for making unbiased estimates of total volume using Cavalieri's principle is described. This is followed by a description of how point probes can be used to make unbiased estimates of the areas of sectional profiles and how, when combined with Cavalieri's principle, point counting can be used to make an unbiased estimate of the volume of a structural feature.
Diversity, distribution and sources of bacteria in residential kitchens
Authors: Flores Gilberto E. , Bates Scott T. , Caporaso J. Gregory , Lauber Christian L. , Leff Jonathan W. , Knight Rob , Fierer Noah
Published: November 2012

Bacteria readily colonize kitchen surfaces, and the exchange of microbes between humans and the kitchen environment can impact human health. However, we have a limited understanding of the overall diversity of these communities, how they differ across surfaces and sources of bacteria to kitchen surfaces. Here we used high‐throughput sequencing of the 16S rRNA gene to explore biogeographical patterns of bacteria across > 80 surfaces within the kitchens of each of four households. In total, 34 bacterial and two archaeal phyla were identified, with most sequences belonging to the Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria. Genera known to contain common food‐borne pathogens were low in abundance but broadly distributed throughout the kitchens, with different taxa exhibiting distinct distribution patterns. The most diverse communities were associated with infrequently cleaned surfaces such as fans above stoves, refrigerator/freezer door seals and floors. In contrast, the least diverse communities were observed in and around sinks, which were dominated by biofilm‐forming Gram‐negative lineages. Community composition was influenced by conditions on individual surfaces, usage patterns and dispersal from source environments. Human skin was the primary source of bacteria across all kitchen surfaces, with contributions from food and faucet water dominating in a few specific locations. This study demonstrates that diverse bacterial communities are widely distributed in residential kitchens and that the composition of these communities is often predictable. These results also illustrate the ease with which human‐ and food‐associated bacteria can be transferred in residential settings to kitchen surfaces.
Extending the Foundations and Reach of Design Science: Further Reflections on the Role of Critical Realism
Authors: Hodgkinson Gerard P. , Starkey Ken
Published: November 2012

In his commentary on our proposal to reframe the relevance of business and management research by combining design science with critical realism (Hodgkinson and Starkey, British Journal of Management, 22 (2011), pp. 355–369), Willmott (British Journal of Management, 23 (2012), pp. 598–604) commends us for our advocacy of a social science as opposed to professional school agenda, while admonishing us for seeking to underpin the commended design science ethos with critical realism, on the grounds of ontological inconsistency and the risk of diminishing the emancipatory elements of our social usefulness agenda. In reply, we demonstrate how, in highlighting the interplay of multiple generative mechanisms within and between levels of analysis, critical realism is entirely commensurate with the design science approach advocated, and enriches rather than diminishes our overarching agenda to humanize the workplace.
Role of new radiation techniques in the treatment of pleural mesothelioma
Authors: Amichetti Maurizio , Lorentini Stefano , Tonoli Sandro , Magrini Stefano Maria
Published: November 2012

Malignant pleural mesothelioma (MPM) is an aggressive neoplasm arising from the surface serosal cells of the pleural cavity. Surgery remains the main therapeutic standard in the treatment of MPM with the goal of complete gross cytoreduction of the tumor. Because MPM is a diffuse disease affecting the entire mesothelial lining of the hemithorax, surgery alone can rarely achieve adequate tumor‐free resection margins. The surgical choices are pleurectomy/decortication (P/D) or extrapleural pneumonectomy (EPP). Radiotherapy (RT) is usually applied postoperatively with the aim to improve local control. However, the efficacy of RT is limited by the large volume of the target to be irradiated (tumor and pleural cavity) and the radiosensitivity of the nearby organs (heart, liver, lung, spinal cord, and esophagus). These factors have historically limited the effective radiation doses that can be given to the patient. There is no role for radical RT alone, but the role of RT as part of multimodality therapy is discussed. After EPP adjuvant RT to the entire hemithorax can reduce the recurrence rate and is well tolerated if strict limits to the dose to contralateral lung are applied: the V20 and V5 (the percent volume of the lung receiving more than 20Gy and 5Gy of radiation) correlate with increased lung toxicity. The use of modern sophisticated techniques allows good target coverage, more conformal high dose delivery, and clinically relevant normal tissue sparing.
Early Life Adversity Contributes to Impaired Cognition and Impulsive Behavior: Studies from the Oklahoma Family Health Patterns Project
Authors: Lovallo William R. , Farag Noha H. , Sorocco Kristen H. , Acheson Ashley , Cohoon Andrew J. , Vincent Andrea S.
Published: November 2012

Background

Stressful early life experience may have adverse consequences in adulthood and may contribute to behavioral characteristics that increase vulnerability to alcoholism. We examined early life adverse experience in relation to cognitive deficits and impulsive behaviors with a reference to risk factors for alcoholism.

Methods

We tested 386 healthy young adults (18 to 30 years of age; 224 women; 171 family history positive for alcoholism) using a composite measure of adverse life experience (low socioeconomic status plus personally experienced adverse events including physical and sexual abuse and separation from parents) as a predictor of performance on the Shipley Institute of Living scale, the Stroop color‐word task, and a delay discounting task assessing preference for smaller immediate rewards in favor of larger delayed rewards. Body mass index (BMI) was examined as an early indicator of altered health behavior.

Results

Greater levels of adversity predicted higher Stroop interference scores (F = 3.07, p = 0.048), faster discounting of delayed rewards (F = 3.79, p = 0.024), lower Shipley mental age scores (F = 4.01, p = 0.019), and higher BMIs in those with a family history of alcoholism (F = 3.40, p = 0.035). These effects were not explained by age, sex, race, education, or depression.

Conclusions

The results indicate a long‐term impact of stressful life experience on cognitive function, impulsive behaviors, and early health indicators that may contribute to risk in persons with a family history of alcoholism.
A prospective study of pregnancy weight gain in Australian women
Authors: de Jersey Susan J. , Nicholson Jan. M. , Callaway Leonie K. , Daniels Lynne A.
Published: November 2012

Background

While weight gain during pregnancy is regarded as important, there has not been a prospective study of measured weight gain in pregnancy in Australia. This study aimed to prospectively evaluate pregnancy‐related weight gain against the Institute of Medicine (IOM) recommendations in women receiving antenatal care in a setting where ongoing weight monitoring is not part of routine clinical practice, to describe women's knowledge of weight gain recommendations and to describe the health professional advice received relating to gestational weight gain (GWG).

Methods

Pregnant women were recruited ≤20 weeks of gestation (n = 664) from a tertiary obstetric hospital between August 2010 to July 2011 for this prospective observational study. Outcome measures were weight gain from pre‐pregnancy to 36 weeks of gestation, weight gain knowledge and health professional advice received.

Results

Thirty‐six percent of women gained weight according to guidelines. Twenty‐six percent gained inadequate weight, and 38% gained excess weight. Fifty‐six percent of overweight women gained weight in excess of the IOM guidelines compared with 30% of those who started with a healthy weight (P < 0.001). At 16 weeks, 47% of participants were unsure of the weight gain recommendations for them. Sixty‐two percent of women reported that the health professionals caring for them during this pregnancy ‘never’ or ‘rarely’ offered advice about how much weight to gain.

Conclusions

The prevalence of inappropriate gestational weight gain in this study was high. The majority of women do not know their recommended weight gain. The advice women received from health professionals relating to healthy weight gain in pregnancy could be improved.
The case for retaining severe perineal tears as an indicator of the quality of obstetric care
Authors: Baghurst Peter A.
Published: November 2012

Background

The incidence of severe perineal tears acquired during vaginal childbirth varies considerably across hospitals but its use as a safety and quality indicator is in jeopardy because of problems associated with accurate detection and recording.

Aim

To understand and interpret time trends in the incidence of third‐ and fourth‐degree perineal tears among women giving birth vaginally in 18 public maternity hospitals in South Australia, taking into account individualised risk factors for each birth.

Methods

The risk‐adjusted probability of a third‐ and fourth‐degree tear was estimated for each of 65,598 singleton vaginal births (2002–2008), using a previously published regression model. The risk factors for each birth included maternal age; parity and ethnicity; assistance with instruments and episiotomy; shoulder dystocia; and infant birthweight. Plots of ‘excess’ tears were generated to help identify maternity services where the observed incidence of severe trauma differed from the expectation estimated from the risk profiles.

Results

Three hospitals were identified at which there were systematically more tears than expected (given their risk profiles), and five hospitals were identified at which there were fewer tears. However, increased tearing at two hospitals coincided closely with improved advocacy for better detection and treatment of perineal tears (especially partial third‐degree tears).

Conclusion

Statistical process control methods provide a powerful means of investigating temporal variations in the incidence of outcomes like severe perineal tears. Third‐ or fourth‐ degree tears should be retained as a quality indicator of maternity services, but it is likely that many third‐degree tears currently go undetected.
Practical recommendations for patient blood management and the reduction of perioperative transfusion in joint replacement surgery
Authors: Bruce Warwick , Campbell David , Daly David , Isbister James
Published: November 2012

Data from the Australian Better Safer Transfusion programme show that about one‐third of patients undergoing hip or knee arthroplasty receive perioperative blood transfusions, placing them at increased risk for adverse clinical outcomes. Other concerns associated with allogeneic blood transfusion include the quality of stored red cell concentrates, the cost of provision of blood and the predicted local demographics, which mean that fewer donors will need to support a greater number of recipients. In view of the multiple challenges associated with allogeneic blood transfusion and its provision, we developed practical management recommendations for perioperative bleeding in joint replacement surgery, based on available evidence and expert consensus opinion, that aim to promote a new, responsible approach to transfusion management. Key recommendations are as follows. Patients' medical health, including haemoglobin and iron levels, needs to be evaluated and optimized preoperatively. Anticoagulant and antiplatelet therapy should be stopped if possible, unless indicated for secondary cardiovascular prevention or coronary stent patency, in which case careful consideration is required. If substantial blood loss is anticipated, intraoperative management with antifibrinolytic agents is recommended for bleeding prophylaxis. Normothermia should be maintained. Pharmacological and non‐pharmacological measures are recommended for post‐operative thromboprophylaxis. A blood management programme should be instituted for haemodynamically stable patients.
Dietary patterns of children at 3.5 and 7 years of age: a New Zealand birth cohort study
Authors: Wall Clare R , Thompson John MD , Robinson Elizabeth , Mitchell Edwin A
Published: November 2012

Aim

To describe the dietary patterns of children at 3.5 and 7 years born small for gestational age (SGA) and appropriate for gestational age (AGA) and the association of dietary patterns with socio‐demographic and obstetric factors.

Methods

Children from a New Zealand birth cohort study were followed up at 3.5 (n = 550) and 7 (n = 591) years. Dietary information was collected using a Food Frequency Questionnaire.

Results

Three dietary patterns were defined in these children (‘traditional’, ‘junk’ and ‘healthy’). Factors associated with dietary patterns were examined in multivariable analyses. At 3.5 years, ‘junk’ was associated with maternal smoking in pregnancy, no attendance at antenatal classes, maternal BMI and a younger maternal age. At 3.5 years, ‘healthy’ was associated with being married when pregnant. At 7 years, ‘traditional’ was associated with being born SGA, smoking in pregnancy, a younger maternal age and being male.

Conclusion

Maternal socio‐demographic, obstetric factors and birth size were associated with dietary patterns. The associations were not consistent with each dietary pattern across age groups. To examine the impact of diet in childhood on growth and health outcomes, factors influencing dietary patterns should be assessed at regular intervals throughout childhood.
Randomised clinical trials: linaclotide phase 3 studies in IBS‐C – a prespecified further analysis based on European Medicines Agency‐specified endpoints
Authors: Quigley E. M. M. , Tack J. , Chey W. D. , Rao S. S. , Fortea J. , Falques M. , Diaz C. , Shiff S. J. , Currie M. G. , Johnston J. M.
Published: November 2012

Background

Treatment options that improve overall symptoms of irritable bowel syndrome with constipation (IBS‐C) are lacking.

Aim

A prespecified further analysis to evaluate the efficacy and safety of linaclotide, a guanylate cyclase C agonist, in patients with IBS‐C, based on efficacy parameters prespecified for European Medicines Agency (EMA) submission.

Methods

Two randomised, double‐blind, multicentre Phase 3 trials investigated once‐daily linaclotide (290 μg) for 12 weeks (Trial 31) or 26 weeks (Trial 302) in patients with IBS‐C. Prespecified primary endpoints were the EMA‐recommended co‐primary endpoints: (i) 12‐week abdominal pain/discomfort responders [≥30% reduction in mean abdominal pain and/or discomfort score (11‐point scales), with neither worsening from baseline, for ≥6 weeks] and (ii) 12‐week IBS degree‐of‐relief responders (symptoms ‘considerably’ or ‘completely’ relieved for ≥6 weeks).

Results

Overall, 803 (Trial 31) and 805 patients (Trial 302) were randomised. A significantly greater proportion of linaclotide‐treated vs. placebo‐treated patients were 12‐week abdominal pain/discomfort responders (Trial 31: 54.8% vs. 41.8%; Trial 302: 54.1% vs. 38.5%; P < 0.001) and IBS degree‐of‐relief responders (Trial 31: 37.0% vs. 18.5%; Trial 302: 39.4% vs. 16.6%; P < 0.0001). Similarly, significantly more linaclotide‐ vs. placebo‐treated patients were responders for ≥13 weeks in Trial 302 (abdominal pain/discomfort: 53.6% vs. 36.0%; IBS degree‐of‐relief: 37.2% vs. 16.9%; P < 0.0001). The proportion of sustained responders (co‐primary endpoint responders plus responders for ≥2 of the last 4 weeks of treatment) was also significantly greater with linaclotide vs. placebo in both trials (P < 0.001).

Conclusion

Linaclotide treatment significantly improved abdominal pain/discomfort and degree‐of‐relief of IBS‐C symptoms compared with placebo over 12 and 26 weeks. Trial registration: ClinicalTrials.gov (identifiers: NCT00948818 and NCT00938717).
Diagnostic value of JAK2 V617F somatic mutation for myeloproliferative cancer in 49 488 individuals from the general population
Authors: Nielsen Camilla , Birgens Henrik S. , Nordestgaard Børge G. , Bojesen Stig E.
Published: November 2012

The JAK2 V617F somatic mutation is present in the majority of patients with myeloproliferative cancer (polycythaemia vera, essential thrombocytosis, and primary myelofibrosis). However, the diagnostic value of the JAK2 V617F somatic mutation for myeloproliferative cancer in the general population is unknown. We examined this question in 49 488 individuals from the Copenhagen General Population Study. We also examined the association between JAK2 V617F somatic mutation, rs10974944 germline genotype, haematological phenotype, any cancer, haematological cancer, myeloproliferative cancer, ischaemic heart disease, and venous thromboembolism. The JAK2 V617F somatic mutation was present in 0·1% (n = 68), increasing across rs10974944 germline genotypes (P‐trend = 0·001). JAK2 V617F somatic mutation positives versus negatives had higher erythrocyte (P = 2 × 10−5), thrombocyte (P = 2 × 10−16), and leucocyte (P = 4 × 10−9) counts, and had 2·7‐/2·5‐fold risk of cancer (prevalent/incident), 44‐/28‐fold risk of haematological cancer, 221‐/97‐fold risk of myeloproliferative cancer, 2·2‐/1·2‐fold risk of ischaemic heart disease, and 3·1‐/1·0‐fold risk of venous thromboembolism. By combining conventional haematological parameters with a test for the JAK2 V617F somatic mutation, myelo;?>proliferative cancer could be identified or ruled out with a sensitivity of 47–100% and a specificity of 98–100%. In conclusion, in the general population the JAK2 V617F somatic mutation has a high diagnostic value for myeloproliferative cancer when combined with conventional haematological parameters.
The effects of autonomous and controlled regulation of performance‐approach goals on well‐being: A process model
Authors: Gillet Nicolas , Lafrenière Marc‐André K. , Vallerand Robert J. , Huart Isabelle , Fouquereau Evelyne
Published: November 2012

The main purpose of the present research was to propose and test a motivational model linking achievement goal approach and self‐determination theory. First, the effects of performance‐approach goals and the autonomous and controlling reasons underlying their pursuit on well‐being were investigated. Second, the mediating variables (i.e., effort, goal attainment, need satisfaction, and thwarting) at play in these relationships were examined based on the self‐concordance model (Sheldon & Elliot, 1999). The model was tested in two studies in educational and work settings using cross sectional (Study 1) and prospective designs (Study 2). The present results revealed that considering autonomous and controlled regulations underlying performance‐approach goals predicted well‐being above and beyond the strength of performance‐approach goals. Moreover, the mediational sequence based on the self‐concordance model was supported in both studies. Theoretical implications and directions for future research are discussed.
The sphingosine kinase inhibitor 2‐(p‐hyroxyanilino)‐4‐(p‐chlorophenyl)thiazole reduces androgen receptor expression via an oxidative stress‐dependent mechanism
Authors: Tonelli Francesca , Alossaimi Manal , Williamson Leon , Tate Rothwelle J , Watson David G , Chan Edmond , Bittman Robert , Pyne Nigel J , Pyne Susan
Published: November 2012

Background and Purpose

Sphingosine kinase catalyses the formation of sphingosine 1‐phosphate and is linked with androgen receptor signalling in prostate cancer cells. Therefore, we investigated the effect of sphingosine kinase inhibitors on androgen receptor expression.

Experimental Approach

Androgen‐sensitive LNCaP cells were treated with SKi (2‐(p‐hydroxyanilino)‐4‐(p‐chlorophenyl)thiazole), which inhibits sphingosine kinases 1 and 2 activity, and the effect on androgen receptor expression was measured.

Key Results

Treatment of cells with SK1 inhibitors reduced the expression of the androgen receptor and prostate‐specific antigen, while (R)‐FTY720 methyl ether (a sphingosine‐kinase‐2‐selective inhibitor), at a concentration that eliminates sphingosine kinase 2 from cells, had no significant effect on androgen receptor expression. The effect of SKi on androgen receptor expression was independent of the SKi‐induced proteasomal degradation of SK1 and was post translational, although androgen receptor mRNA transcript was reduced. Fumonisin B1 (a ceramide synthase inhibitor) also failed to reverse the effect of SKi on androgen receptor expression, thereby excluding a role for ceramide derived from the salvage pathway. The effect of SKi on androgen receptor expression was reversed by N‐acetylcysteine, which was used to scavenge reactive oxygen species.

Conclusion and Implications

Inhibition of sphingosine kinase 1 activity abrogates androgen receptor signalling via an oxidative stress‐induced, p53‐independent mechanism in prostate cancer cells. Therefore, SK1 inhibitors may offer therapeutic potential in promoting the removal of AR receptors from prostate cancer cells, resulting in an increased efficacy, which is likely to be superior to inhibitors that simply reversibly inhibit AR signalling.
Glutathione S‐transferase P1‐1 as a target for mesothelioma treatment
Authors: De Luca Anastasia , Pellizzari Tregno Francesca , Sau Andrea , Pastore Anna , Palumbo Camilla , Alama Angela , Cicconi Rosella , Federici Giorgio , Caccuri Anna M.
Published: November 2012

Malignant pleural mesothelioma is a poorly responsive tumor known to overexpress the phase II detoxification enzyme glutathione‐S‐transferase, which catalyzes the conjugation between glutathione and platinum(II)‐containing drugs. Therefore, we evaluated the effect of the strong glutathione S‐transferase inhibitor NBDHEX on human mesothelioma cell lines (MSTO‐211H, MPP89, MM‐B1 and Mero 48a) featuring the most common mesothelioma phenotypes: epithelioid and biphasic. Even though a different response to NBDHEX was observed, the molecule was very effective on all cell lines tested, triggering a sustained activation of both JNK and p38, followed by caspase activation and apoptosis. NBDHEX also caused severe oxidative stress in the MPP89 cells and, to a lesser extent, in the MMB1 cells, while it did not cause a significant redox imbalance in the other cell lines. The efficacy of the drug was found to be comparable or even higher than that of cisplatin. Moreover, it showed synergistic or additive effects when used in combination with cisplatin. In conclusion, NBDHEX was effective on mesothelioma cell lines, with IC50 values in the low micromolar range (IC50 between 1 and 4 μM). These findings indicate that NBDHEX, alone or in combination with cisplatin, is a promising new strategy for treating this rare and aggressive malignancy.
Characteristics ofin-vitrophenotypes of glutamic acid decarboxylase 65 autoantibodies in high-titre individuals
Authors: Chéramy M , Hampe C S , Ludvigsson J , Casas R
Published: November 2012

Previous studies have indicated phenotypical differences in glutamic acid decarboxylase 65 autoantibodies (GADA) found in type 1 diabetes (T1D) patients, individuals at risk of developing T1D and stiff-person syndrome (SPS) patients. In a Phase II trial using aluminium-formulated GAD65 (GAD-alum) as an immunomodulator in T1D, several patients responded with high GADA titres after treatment, raising concerns as to whether GAD-alum could induce GADA with SPS-associated phenotypes. This study aimed to analyse GADA levels, immunoglobulin (Ig)G1–4 subclass frequencies, b78- and b96·11-defined epitope distribution and GAD65 enzyme activity in sera from four cohorts with very high GADA titres: T1D patients (n = 7), GAD-alum-treated T1D patients (n = 9), T1D high-risk individuals (n = 6) and SPS patients (n = 12). SPS patients showed significantly higher GADA levels and inhibited the in-vitro GAD65 enzyme activity more strongly compared to the other groups. A higher binding frequency to the b78-defined epitope was found in the SPS group compared to T1D and GAD-alum individuals, whereas no differences were detected for the b96·11-defined epitope. GADA IgG1–4 subclass levels did not differ between the groups, but SPS patients had higher IgG2 and lower IgG4 distribution more frequently. In conclusion, the in-vitro GADA phenotypes from SPS patients differed from the T1D- and high-risk groups, and GAD-alum treatment did not induce SPS-associated phenotypes. However, occasional overlap between the groups exists, and caution is indicated when drawing conclusions to health or disease status.
Treatment of soft tissue recessions at titanium implants using a resorbable collagen matrix: a pilot study
Authors: Schwarz Frank , Mihatovic Ilja , Shirakata Yoshinori , Becker Jürgen , Bosshardt Dieter , Sculean Anton
Published: November 2012

Objectives

To histologically assess the effectiveness of a porcine‐derived collagen matrix (CM) and a subepithelial connective tissue graft (CTG) for the coverage of single mucosal recessions at osseointegrated dental implants.

Materials and methods

Chronic‐type mucosal Miller Class I‐like recessions (mean clinical defect height: 0.67 ± 0.33–1.16 ± 0.19 mm) were established at the buccal aspect of titanium implants with platform switch in six beagle dogs. The defects were randomly allocated to either (1) coronally advanced flap surgery (CAF) + CM, (2) CAF + CTG or (3) CAF alone. At 12 weeks, histomorphometrical measurements were made (e.g.) between the implant shoulder (IS) and the mucosal margin (PM) and IS and the outer contour of the adjacent soft tissue (mucosal thickness [MT]).

Results

All treatment procedures investigated were associated with an almost complete soft tissue coverage of the defect area (i.e. coronal positioning of PM relative to IS). Mean IS‐PM and MT values tended to be increased in both CAF + CM (1.04 ± 0.74 mm/0.71 ± 0.55 mm) and CAF + CTG (0.88 ± 1.23 mm/0.62 ± 0.66 mm) groups when compared with CAF (0.16 ± 0.28 mm/0.34 ± 0.23 mm) alone. These differences, however, did not reach statistical significance.

Conclusions

Within the limits of this pilot study, it was concluded that all treatment procedures investigated were effective in covering soft tissue recessions at titanium implants.
Diagnostic utility of direct immunofluorescence findings around hair follicles and sweat glands in immunobullous disease
Authors: Lehman Julia S. , Camilleri Michael J.
Published: November 2012

Background

Inherent to some immunobullous disorders is potential for intraepidermal or dermal–epidermal junction fragility, a phenomenon that may compromise biopsy specimen integrity and direct immunofluorescence (DIF) interpretation. In these situations, cutaneous adnexal structures (e.g. hair follicles, sweat apparatus) usually remain intact. Whether periadnexal DIF findings are reliable in diagnosing immunobullous conditions is unknown.

Methods

We evaluated 56 cutaneous specimens with diagnostic immunoglobulin (Ig) deposition patterns that contained adnexal structures. In a corollary study, we examined 145 hematoxylin‐eosin‐stained frozen specimens to determine biopsy factors associated with the presence of adnexal structures.

Results

Periadnexal DIF findings offered diagnostic sensitivity in conditions with linear or cell‐surface Ig deposition or lupus band. Periadnexal DIF findings were unreliable in dermatitis herpetiformis. Biopsy specimens from scalp and genitalia were most likely to contain folliculosebaceous units and sweat duct apparatus, respectively. Relative depth of biopsy correlated directly with the likelihood of identifying sweat duct apparatus but not folliculosebaceous units.

Conclusions

Periadnexal DIF findings may add diagnostic sensitivity in DIF evaluation of pemphigoid, pemphigus and lupus erythematosus. Pathologists can guide clinicians to biopsy certain anatomic sites and to obtain sufficient biopsy depth to increase the probability of capturing adnexal structures and, therefore, diagnostic yield from DIF specimens.
Increased hepatitis C virus vaccine clinical trial literacy following a brief intervention among people who inject drugs
Authors: White Bethany , Madden Annie , Hellard Margaret , Kerr Thomas , Prins Maria , Page Kimberly , Dore Gregory J. , Maher Lisa
Published: November 2012

Introduction and Aims

While people who inject drugs are at high risk of hepatitis C virus (HCV) infection and will be the target population for future HCV vaccine trials, little is known about clinical trial literacy (CTL) in this group. We assessed the impact of a brief intervention (BI) designed to improve HCV vaccine CTL among people who inject drugs in Sydney, Australia.

Design and Methods

People who inject drugs enrolled in a community‐based prospective observational study between November 2008 and September 2010 (n = 102) completed a CTL assessment followed immediately by the BI. Post‐test assessment was conducted at 24 weeks.

Results

The median age of the sample was 27 years, 73% were male and 60% had 10 or less years of schooling. The median time since first injection was 5 years and 20% reported daily or more frequent injecting. The mean number of correct responses increased from 5.3 to 6.3/10 (t = −4.2; 101df, P < 0.001) 24 weeks post‐intervention. Statistically significant differences were observed for three knowledge items with higher proportions of participants correctly answering questions related to randomisation (P = 0.002), blinding (P = 0.005) and vaccine‐induced seropositivity (P = 0.003) post‐intervention.

Discussion and Conclusions

A significant increase in HCV vaccine CTL was observed, suggesting that new and relatively novel concepts can be learned and recalled in this group. These findings support the feasibility of future trials among this population. [Correction added on 21 November 2012, after first online publication: T‐score for mean number of correct responses was corrected to ‘−4.2’ in the Results section.] [White B, Madden A, Hellard H, Kerr T, Prins M, Page K, Dore GJ, Maher L. Increased hepatitis C virus vaccine clinical trial literacy following a brief intervention among people who inject drugs. Drug Alcohol Rev 2013;32:419–425]
Energy loss via urine and faeces – a combustive analysis in diabetic rats and the impact of antidiabetic treatment on body weight
Authors: Elvert R. , Wille A. , Wandschneider J. , Werner U. , Glombik H. , Herling A. W.
Published: November 2012

Aims

Intensive glycaemic control in type 2 diabetes achieved by insulin is generally accompanied by body weight gain. This study was performed to emphasize the meaning of caloric analysis of urine and faeces for energy balance.

Methods

We measured energetic loss via urine and faeces during antihyperglycaemic treatment in male obese Zucker diabetic fatty (ZDF) rats. Rats were treated for 10 days with the sodium–glucose‐linked transporter‐2 (SGLT2) inhibitor AVE2268, with insulin glargine, with the GLP‐1 receptor agonist lixisenatide and with the combination of insulin glargine and lixisenatide. Each study was accompanied by one lean (Fa/?) and one obese (fa/fa) untreated non‐diabetic and diabetic control group, respectively. Blood glucose, body weight alterations and food assimilation efficiency were monitored.

Results

In control ZDF rats, more than 12 g/day of pure glucose was urinarily excreted. In total, the energetic loss via urine exceeded 30% from total energy uptake. Insulin glargine treatment decreased urinary energetic loss, leading to a body weight gain of approximately 3 g/day. An almost body weight‐neutral antihyperglycaemic treatment could be achieved with AVE2268 and lixisenatide. While lixisenatide reduced body weight gain via reduction of energy uptake, the SGLT2 inhibitor even increased urinary glucose and thus energy excretion. Combining insulin glargine with lixisenatide attenuated the anabolic effect of insulin resulting in weight neutrality.

Conclusions

Our data clearly show renal contribution to the body's energy control by urinary glucose excretion (UGE) during antidiabetic treatment. The undesired retained energy could be reduced via additional UGE or via simultaneous reduction of energy uptake and/or energy retention.
Response to pretransplant hypomethylating agents influences the outcome of allogeneic hematopoietic stem cell transplantation in adults with myelodysplastic syndromes
Authors: Yahng Seung‐Ah , Yoon Jae‐Ho , Shin Seung‐Hwan , Lee Sung‐Eun , Cho Byung‐Sik , Lee Dong‐Gun , Eom Ki‐Seong , Lee Seok , Min Chang‐Ki , Kim Hee‐Je , Cho Seok‐Goo , Kim Dong‐Wook , Lee Jong‐Wook , Min Woo‐Sung , Kim Tai‐Gyu , Park Chong‐Won , Kim Yoo‐Jin
Published: November 2012

This study describes a retrospective analysis on the transplant outcome of 56 consecutive patients with myelodysplastic syndrome (MDS) according to their response to hypomethylating agents (HMA). While 2‐yr disease‐free survival (DFS) of patients who transformed to acute myeloid leukemia (n = 12) was 25%, that of the remaining patients with MDS according to response to HMA was 73.1%, 68.1%, 50.0%, and 20.8% in G‐COR (group of continuous response, n = 19), G‐NoC (group of no change, n = 15), G‐LOR (group of loss of response, n = 6), and G‐DP (group of disease progression, n = 4), respectively. When dichotomized as G‐COR/G‐NoC versus G‐LOR/G‐DP, significantly different 2‐yr DFS (71.0% vs. 33.3%; P = 0.004) and relapse (14.1% vs. 46.7%; P = 0.016) were demonstrated. On multivariate analysis, G‐LOR/G‐DP [hazard ratio (HR), 3.91; P = 0.008] and poor karyotype at transplantation (HR, 2.69; P = 0.017) were the significant predictors for poor DFS, as G‐LOR/G‐DP was for relapse (HR, 6.28; P = 0.011). DFS was significantly poor in patients with any of the two predictors in all MDS (81.5% vs. 34.9%; P = 0.001) or higher‐risk MDS (HrMDS) at the time of HMA (80.7% vs. 29.2%; P = 0.005). G‐COR showed a trend of better DFS compared with G‐NoC among HrMDS (74.6% vs. 36.5%; P = 0.090). These results implicate the significance of response to HMA on hematopoietic stem cell transplantation (HSCT) outcomes and support the need for future study to verify the suggested strategy of proceeding to transplantation before LOR or DP, especially for HrMDS.
The effect of auditory cortex deactivation on stimulus‐specific adaptation in the inferior colliculus of the rat
Authors: Anderson L. A. , Malmierca M. S.
Published: November 2012

Many neurons in the central auditory pathway, from the inferior colliculus (IC) to the auditory cortex (AC), respond less strongly to a commonly occurring stimulus than one that rarely occurs. The origin of this phenomenon, called stimulus‐specific adaptation (SSA), remains uncertain. The AC sends descending projections to the IC that terminate most densely upon the dorsal, lateral and rostral IC cortices – areas where strong SSA has been reported. To investigate whether SSA in the IC is dependent upon the AC for its generation, we recorded the response from single IC neurons to stimuli presented in an oddball paradigm before, during and after reversibly deactivating the ipsilateral AC with a cryoloop. While changes in the basic response properties of the IC neurons were widespread (89%), changes in SSA sensitivity were less common; approximately half of the neurons recorded showed a significant change in SSA, while the other half remained unchanged. Changes in SSA could be in either direction: 18% enhanced their SSA sensitivity, while 34% showed reduced SSA sensitivity. For the majority of this latter group, cortical deactivation reduced, but did not eliminate, significant SSA levels. Only eight neurons seemed to inherit SSA from the AC, as their pre‐existing significant level of SSA became non‐significant during cortical deactivation. Thus, the presence of SSA in the IC is generally not dependent upon the corticocollicular projection, suggesting the AC is not essential for the generation of subcortical SSA; however, the AC may play a role in the modulation of subcortical SSA.
The effects of Sophora angustifolia and other natural plant extracts on melanogenesis and melanin transfer in human skin cells
Authors: Singh Suman K. , Baker Richard , Wibawa Judata I. D. , Bell Mike , Tobin Desmond J.
Published: November 2012

Skin pigmentation is a multistep process of melanin synthesis by melanocytes, its transfer to recipient keratinocytes and its degradation. As dyspigmentation is a prominent marker of skin ageing, novel effective agents that modulate pigmentation safely are being sought for both clinical and cosmetic use. Here, a number of plant extracts were examined for their effect on melanogenesis (by melanin assay and Western blotting) and melanin transfer (by confocal immunomicroscopy of gp100‐positive melanin granules in cocultures and by SEM analysis of filopodia), in human melanocytes and in cocultures with phototype‐matched normal adult epidermal keratinocytes. Mulberry, Kiwi and Sophora extracts were assessed against isobutylmethylxanthine, hydroquinone, vitamin C and niacinamide. Compared with unstimulated control, all extracts significantly reduced melanogenesis in human melanoma cells and normal adult epidermal melanocytes. These extracts also reduced melanin transfer and reduced filopodia expression on melanocytes, similar to hydroquinone and niacinamide, indicating their effectiveness as multimode pigmentation actives.
Therapeutic drug monitoring of imatinib in chronic myeloid leukemia: experience from 1216 patients at a centralized laboratory
Authors: Bouchet Stéphane , Titier Karine , Moore Nicholas , Lassalle Régis , Ambrosino Basmah , Poulette Sylvie , Schuld Peter , Belanger Coralie , Mahon François‐Xavier , Molimard Mathieu
Published: November 2012

This study set out to examine in a large real‐life cohort of patients with chronic myeloid leukemia (CML) the impact of imatinib threshold of 1000 ng/mL on molecular response, as suggested in a small subset of patients. Patient plasma samples were submitted from around France to a central facility, free of charge under the auspices of the European Treatment and Outcome Study (EUTOS) for CML. Submitting physicians were required to complete an ‘imatinib monitoring request form’, including details of why therapeutic drug monitoring (TDM) was requested, dose and duration of imatinib treatment, cytogenetic and molecular response, adverse events, and concurrent medications. Imatinib trough plasma concentration (Cmin) was measured at the central facility. Among 1985 eligible plasma samples analyzed, from 1216 CML patients, imatinib Cmin correlated positively with reported imatinib dose, but interpatient variability in Cmin was high (60%). A logistic regression analysis revealed that treatment duration and imatinib Cmin > 1000 ng/mL were significantly associated with major and complete molecular responses with odds ratios of 1.69 and 2.08, respectively. These data support in real‐life setting that imatinib Cmin threshold of 1000 ng/mL is associated with major and complete molecular response and that TDM could play an important role in dose optimization.
Genetic and phenotypic variation among Galaxias maculatus populations reflects contrasting landscape effects between northern and southern Patagonia
Authors: CARREA CECILIA , CUSSAC VICTOR E. , RUZZANTE DANIEL E.
Published: November 2012

1. Understanding the influence of landscape characteristics on genetic and phenotypic intraspecific variability can yield insights into how evolutionary processes work as well as provide essential information for the conservation of biodiversity.

2. Our aim was to compare the genetic structure and phenotypic variation among Galaxias maculatus populations inhabiting two Atlantic Ocean river basins in Patagonia in relation to historical and contemporary landscape characteristics associated with latitude.

3. Population genetic analysis (based on eight microsatellite loci, 505 individuals) indicates that genetic structure is more pronounced in the Río Negro basin (RN) at 39–41°S (10 localities) than in the southernmost Santa Cruz River basin (SCR) at 49–50°S (seven localities). Spatial autocorrelation analysis showed different effects of geographic distance in shaping population differentiation patterns in the two basins.

4. While in northern Patagonia, fragmented populations could have survived the severity of Quaternary climate cycles, at higher latitudes G. maculatus populations were probably extirpated by extensive ice sheets. Extant populations in the upper reaches of the southernmost basin probably originated from refugia close to the Atlantic Ocean.

5. Based on otolith microchemistry, we have documented facultative diadromy for the first time for the species in the SCR basin, while no evidence of migration to the sea was found in individuals from the RN basin. Vertebral number increased with latitude, and within the SCR basin, the higher counts are possibly associated with a migratory life style.