bluuebunny/crossref_metadata_2025 huggingface dataset:
Speech recognition software and electronic psychiatric progress notes: physicians' ratings and preferences
Authors: Derman Yaron D , Arenovich Tamara , Strauss John
Published: August 2010

Background

The context of the current study was mandatory adoption of electronic clinical documentation within a large mental health care organization. Psychiatric electronic documentation has unique needs by the nature of dense narrative content. Our goal was to determine if speech recognition (SR) would ease the creation of electronic progress note (ePN) documents by physicians at our institution.

Methods

Subjects: Twelve physicians had access to SR software on their computers for a period of four weeks to create ePN. Measurements: We examined SR software in relation to its perceived usability, data entry time savings, impact on the quality of care and quality of documentation, and the impact on clinical and administrative workflow, as compared to existing methods for data entry. Data analysis: A series of Wilcoxon signed rank tests were used to compare pre- and post-SR measures. A qualitative study design was used.

Results

Six of twelve participants completing the study favoured the use of SR (five with SR alone plus one with SR via hand-held digital recorder) for creating electronic progress notes over their existing mode of data entry. There was no clear perceived benefit from SR in terms of data entry time savings, quality of care, quality of documentation, or impact on clinical and administrative workflow.

Conclusions

Although our findings are mixed, SR may be a technology with some promise for mental health documentation. Future investigations of this nature should use more participants, a broader range of document types, and compare front- and back-end SR methods.
A novel function for vimentin: the potential biomarker for predicting melanoma hematogenous metastasis
Authors: Li Man , Zhang Baogang , Sun Baocun , Wang Xuan , Ban Xinchao , Sun Tao , Liu Zhiyong , Zhao Xiulan
Published: August 2010

Background

The incidence of malignant melanoma (MM) was occurring at a faster rate than for most neoplasm worldwide, and melanoma metastasis is still the most formidable problem. So it is necessarily to find some biomarkers associated with melanoma metastasis.

Methods

In our study, 8 spontaneous lung metastatic mice models were created by B16F10 subcutaneously transplantation. The differential protein profiles of two kinds of subcutaneous transplanted tumor tissues, which was parental B16F10 (B16 group) and corresponding lung metastases (B16M group) were detected by two-dimensional differential in-gel electrophoresis (2D-DIGE) combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Western blotting was used to validate the results, and the clinical significance of individual protein was detected furtherly in a set of human samples.

Result

In this study, thirty proteins were found to be differentially expressed (ratio > 2 or < -2, P < 0.01) and thirteen of them were identified by MS. Highly expressed proteins in B16M group included cytoskeleton/structure proteins (vimentin, gamma-actin, β-actin, laminin binding protein), the chaperone family of proteins (heavy-chain binding protein, Bip), immunoproteasome assembly (proteasome activator REG alpha) and others involved in glycolysis activity (PGK1, enolase, TPI, human skeletal muscle GAPDH) and protein transport (myoglobin). Vimentin was significantly up-regulated in B16M group compared with B16 group which was validated by western blotting. Immunohistochemistry was performed in a set of clinical samples, the results showed that over-expression of vimentin was frequently observed in primary melanoma patients with hematogenous metastasis (P < 0.05), not associated with lymph node metastasis (P > 0.05). The presence of TNM stage was a independent indicator of poor prognosis for melanoma patients (P = 0.004).

Conclusion

The aberrant immunohistochemical expression of vimentin in primary melanoma tissues may help to call attention for patients with high risk of hematogenous metastasis. That might be as a novel metastatic indicator for melanoma. In a word, vimentin is not only the dignostic marker but also the hematogenous metastasis predictor for melanomas clinically.
Immediate determination of ACPA and rheumatoid factor - a novel point of care test for detection of anti-MCV antibodies and rheumatoid factor using a lateral-flow immunoassay
Authors: Renger Franziska , Bang Holger , Feist Eugen , Fredenhagen Gert , Natusch Alexander , Backhaus Marina , Burmester Gerd-R , Egerer Karl
Published: August 2010

Introduction

Autoantibodies against mutated and citrullinated vimentin (MCV) represent a novel diagnostic marker for rheumatoid arthritis (RA). Recently, an increased sensitivity for anti-MCV compared to autoantibodies against cyclic citrullinated peptides (anti-CCP2) was shown in cohorts of patients with early RA and established disease.

The aim of this study was to develop and evaluate a point of care test (POCT) for detection of anti-MCV antibodies immediately at the first visit or at the bed side.

Methods

A lateral-flow immunoassay was developed for simultaneous detection of anti-MCV antibodies and rheumatoid factor (RF-IgG) and evaluated in a prospective setting. Analyses were performed from whole blood samples of patients with seropositive RA (n = 108), seronegative RA as well as other rheumatic disorders (n = 122), and healthy blood donors (n = 200) and compared to detection via ELISA.

Results

Using the POCT, anti-MCV antibodies were detected in 54.6% and RF-IgG in 56.5% of patients with RA. Specificity was 99.1% for anti-MCV antibodies and 91.2% for RF-IgG. Compared to ELISA's results, POCT sensitivity was 69.3% for anti-MCV and 55.6% for RF-IgG, specificity was 99.7% and 97.2%, respectively.

Conclusions

This POCT for detection of anti-MCV antibodies and RF-IgG provides high specificity for the diagnosis of RA and is useful in clinical practice due to its simplicity and its reliable performance. This test can greatly improve a timely management of RA and may help in screening patients with suspected RA in non-specialized settings prompting early referrals.
The effect of risedronate on osteogenic lineage is mediated by cyclooxygenase-2 gene upregulation
Authors: Valenti Maria Teresa , Giannini Sandro , Donatelli Luca , Zanatta Mirko , Bertoldo Francesco , Sella Stefania , Vilei Maria Teresa , Ossi Elena , Realdi Giuseppe , Lo Cascio Vincenzo , Dalle Carbonare Luca
Published: August 2010

Introduction

The purpose of this study was to evaluate the effects of risedronate (Ris) in the modulation of bone formation in rats with glucocorticoid (GC)-induced osteoporosis by histomorphometric, immunohistochemical and gene expression analyses.

Methods

We analyzed structure, turnover and microarchitecture, cyclooxygenase 2 (COX-2) levels and osteocyte apoptosis in 40 female rats divided as follows: 1) vehicle of methylprednisolone (vGC) + vehicle of risedronate (vRis); 2) Ris 5 μg/Kg + vGC; 3) methylprednisolone (GC) 7 mg/Kg + vRis; 4) GC 7 mg/Kg +Ris 5 μg/Kg. In addition, we evaluated cell proliferation and expression of COX-2 and bone alkaline phosphatase (b-ALP) genes in bone marrow cells and MLO-y4 osteocytes treated with Ris alone or in co-treatment with the selective COX-2 inhibitor NS-398 or with dexametasone.

Results

Ris reduced apoptosis induced by GC of osteocytes (41% vs 86%, P < 0.0001) and increased COX-2 expression with respect to controls (Immuno-Hystochemical Score (IHS): 8.75 vs 1.00, P < 0.0001). These positive effects of Ris in bone formation were confirmed by in vitro data as the viability and expression of b-ALP gene in bone marrow cells resulted increased in a dose dependent manner.

Conclusions

These findings suggest a positive effect of Ris in bone formation and support the hypothesis that the up-regulation of COX-2 could be an additional mechanism of anabolic effect of Ris.
Dynamic changes in gene expression in vivo predict prognosis of tamoxifen-treated patients with breast cancer
Authors: Taylor Karen J , Sims Andrew H , Liang Liang , Faratian Dana , Muir Morwenna , Walker Graeme , Kuske Barbara , Dixon J Michael , Cameron David A , Harrison David J , Langdon Simon P
Published: August 2010

Introduction

Tamoxifen is the most widely prescribed anti-estrogen treatment for patients with estrogen receptor (ER)-positive breast cancer. However, there is still a need for biomarkers that reliably predict endocrine sensitivity in breast cancers and these may well be expressed in a dynamic manner.

Methods

In this study we assessed gene expression changes at multiple time points (days 1, 2, 4, 7, 14) after tamoxifen treatment in the ER-positive ZR-75-1 xenograft model that displays significant changes in apoptosis, proliferation and angiogenesis within 2 days of therapy.

Results

Hierarchical clustering identified six time-related gene expression patterns, which separated into three groups: two with early/transient responses, two with continuous/late responses and two with variable response patterns. The early/transient response represented reductions in many genes that are involved in cell cycle and proliferation (e.g. BUB1B, CCNA2, CDKN3, MKI67, UBE2C), whereas the continuous/late changed genes represented the more classical estrogen response genes (e.g. TFF1, TFF3, IGFBP5). Genes and the proteins they encode were confirmed to have similar temporal patterns of expression in vitro and in vivo and correlated with reduction in tumour volume in primary breast cancer. The profiles of genes that were most differentially expressed on days 2, 4 and 7 following treatment were able to predict prognosis, whereas those most changed on days 1 and 14 were not, in four tamoxifen treated datasets representing a total of 404 patients.

Conclusions

Both early/transient/proliferation response genes and continuous/late/estrogen-response genes are able to predict prognosis of primary breast tumours in a dynamic manner. Temporal expression of therapy-response genes is clearly an important factor in characterising the response to endocrine therapy in breast tumours which has significant implications for the timing of biopsies in neoadjuvant biomarker studies.
Withdrawing intra-aortic balloon pump support paradoxically improves microvascular flow
Authors: Munsterman Luuk DH , Elbers Paul WG , Ozdemir Alaattin , van Dongen Eric PA , van Iterson Mat , Ince Can
Published: August 2010

Introduction

The Intra-Aortic Balloon Pump (IABP) is frequently used to mechanically support the heart. There is evidence that IABP improves microvascular flow during cardiogenic shock but its influence on the human microcirculation in patients deemed ready for discontinuing IABP support has not yet been studied. Therefore we used sidestream dark field imaging (SDF) to test our hypothesis that human microcirculation remains unaltered with or without IABP support in patients clinically ready for discontinuation of mechanical support.

Methods

We studied 15 ICU patients on IABP therapy. Measurements were performed after the clinical decision was made to remove the balloon catheter. We recorded global hemodynamic parameters and performed venous oximetry during maximal IABP support (1:1) and 10 minutes after temporarily stopping the IABP therapy. At both time points, we also recorded video clips of the sublingual microcirculation. From these we determined indices of microvascular perfusion including perfused vessel density (PVD) and microvascular flow index (MFI).

Results

Ceasing IABP support lowered mean arterial pressure (74 ± 8 to 71 ± 10 mmHg; P = 0.048) and increased diastolic pressure (43 ± 10 to 53 ± 9 mmHg; P = 0.0002). However, at the level of the microcirculation we found an increase of PVD of small vessels <20 μm (5.47 ± 1.76 to 6.63 ± 1.90; P = 0.0039). PVD for vessels >20 μm and MFI for both small and large vessels were unaltered. During the procedure global oxygenation parameters (ScvO2/SvO2) remained unchanged.

Conclusions

In patients deemed ready for discontinuing IABP support according to current practice, SDF imaging showed an increase of microcirculatory flow of small vessels after ceasing IABP therapy. This observation may indicate that IABP impairs microvascular perfusion in recovered patients, although this warrants confirmation.
Galaxy: a comprehensive approach for supporting accessible, reproducible, and transparent computational research in the life sciences
Authors: Goecks Jeremy , Nekrutenko Anton , Taylor James ,   The Galaxy Team
Published: August 2010

Increased reliance on computational approaches in the life sciences has revealed grave concerns about how accessible and reproducible computation-reliant results truly are. Galaxy http://usegalaxy.org, an open web-based platform for genomic research, addresses these problems. Galaxy automatically tracks and manages data provenance and provides support for capturing the context and intent of computational methods. Galaxy Pages are interactive, web-based documents that provide users with a medium to communicate a complete computational analysis.
The myotubularin phosphatase MTMR4 regulates sorting from early endosomes
Authors: Naughtin Monica J. , Sheffield David A. , Rahman Parvin , Hughes William E. , Gurung Rajendra , Stow Jennifer L. , Nandurkar Harshal H. , Dyson Jennifer M. , Mitchell Christina A.
Published: August 2010

Phosphatidylinositol 3-phosphate [PtdIns(3)P] regulates endocytic trafficking and the sorting of receptors through early endosomes, including the rapid recycling of transferrin (Tfn). However, the phosphoinositide phosphatase that selectively opposes this function is unknown. The myotubularins are a family of eight catalytically active and six inactive enzymes that hydrolyse PtdIns(3)P to form PtdIns. However, the role each myotubularin family member plays in regulating endosomal PtdIns(3)P and thereby endocytic trafficking is not well established. Here, we identify the myotubularin family member MTMR4, which localizes to early endosomes and also to Rab11- and Sec15-positive recycling endosomes. In cells with MTMR4 knockdown, or following expression of the catalytically inactive MTMR4, MTMR4C407A, the number of PtdIns(3)P-decorated endosomes significantly increased. MTMR4 overexpression delayed the exit of Tfn from early endosomes and its recycling to the plasma membrane. By contrast, expression of MTMR4C407A, which acts as a dominant-negative construct, significantly accelerated Tfn recycling. However, in MTMR4 knockdown cells Tfn recycling was unchanged, suggesting that other MTMs might also contribute to recycling. MTMR4 regulated the subcellular distribution of Rab11 and, in cells with RNAi-mediated knockdown of MTMR4, Rab11 was directed away from the pericentriolar recycling compartment. The subcellular distribution of VAMP3, a v-SNARE protein that resides in recycling endosomes and endosome-derived transport vesicles, was also regulated by MTMR4. Therefore, MTMR4 localizes at the interface of early and recycling endosomes to regulate trafficking through this pathway.
Regulation of mixed-lineage kinase activation in JNK-dependent morphogenesis
Authors: Garlena Rebecca A. , Gonda Rebecca L. , Green Alyssa B. , Pileggi Rachel M. , Stronach Beth
Published: August 2010

Normal cells respond appropriately to various signals, while sustaining proper developmental programs and tissue homeostasis. Inappropriate signal reception, response or attenuation, can upset the normal balance of signaling within cells, leading to dysfunction or tissue malformation. To understand the molecular mechanisms that regulate protein-kinase-based signaling in the context of tissue morphogenesis, we analyzed the domain requirements of Drosophila Slpr, a mixed-lineage kinase (MLK), for Jun N-terminal kinase (JNK) signaling. The N-terminal half of Slpr is involved in regulated signaling whereas the C-terminal half promotes cortical protein localization. The SH3 domain negatively regulates Slpr activity consistent with autoinhibition via a conserved proline motif. Also, like many kinases, conserved residues in the activation segment of the catalytic domain regulate Slpr. Threonine 295, in particular, is essential for function. Slpr activation requires dual input from the MAP4K Misshapen (Msn), through its C-terminal regulatory domain, and the GTPase Rac, which both bind to the LZ–CRIB region of Slpr in vitro. Although Rac is sufficient to activate JNK signaling, our results indicate that there are Slpr-independent functions for Rac in dorsal closure. Finally, expression of various Slpr constructs alone or with upstream activators reveals a wide-ranging response at the cell and tissue level.
Growth differentiation factor 9 signaling requires ERK1/2 activity in mouse granulosa and cumulus cells
Authors: Sasseville Maxime , Ritter Lesley J. , Nguyen Thao M. , Liu Fang , Mottershead David G. , Russell Darryl L. , Gilchrist Robert B.
Published: August 2010

Ovarian folliculogenesis is driven by the combined action of endocrine cues and paracrine factors. The oocyte secretes powerful mitogens, such as growth differentiation factor 9 (GDF9), that regulate granulosa cell proliferation, metabolism, steroidogenesis and differentiation. This study investigated the role of the epidermal growth factor receptor (EGFR)–extracellular signal-regulated kinase 1 and 2 (ERK1/2; also known as MAPK3/1) signaling pathway on GDF9 action on granulosa cells. Results show that mitogenic action of the oocyte is prevented by pharmacological inhibition of the EGFR–ERK1/2 pathway. Importantly, EGFR–ERK1/2 activity as well as rous sarcoma oncogene family kinases (SFK) are required for signaling through SMADs, mediating GDF9, activin A and TGFβ1 mitogenic action in granulosa cells. GDF9 could not activate ERK1/2 or affect EGF-stimulated ERK1/2 in granulosa cells. However, induction of the SMAD3-specific CAGA reporter by GDF9 in granulosa cells required active EGFR, SFKs and ERK1/2 as did GDF9-responsive gene expression. Finally, the EGFR–SFKs–ERK1/2 pathway was shown to be required for the maintenance of phosphorylation of the SMAD3 linker region. Together our results suggest that receptivity of granulosa cells to oocyte-secreted factors, including GDF9, is regulated by the level of activation of the EGFR and resulting ERK1/2 activity, through the requisite permissive phosphorylation of SMAD3 in the linker region. Our results indicate that oocyte-secreted TGFβ-like ligands and EGFR–ERK1/2 signaling are cooperatively required for the unique granulosa cell response to the signal from oocytes mediating granulosa cell survival and proliferation and hence the promotion of follicle growth and ovulation.
The fibroblast-derived paracrine factor neuregulin-1 has a novel role in regulating the constitutive color and melanocyte function in human skin
Authors: Choi Wonseon , Wolber Rainer , Gerwat Wolfram , Mann Tobias , Batzer Jan , Smuda Christoph , Liu Hongfang , Kolbe Ludger , Hearing Vincent J.
Published: August 2010

Interactions between melanocytes and neighboring cells in the skin are important in regulating skin color in humans. We recently demonstrated that the less pigmented and thicker skin on the palms and soles is regulated by underlying fibroblasts in those areas, specifically via a secreted factor (DKK1) that modulates Wnt signaling. In this study, we tested the hypothesis that dermal fibroblasts regulate the constitutive skin color of individuals ranging from very light to very dark. We used microarray analysis to compare gene expression patterns in fibroblasts derived from lighter skin types compared to darker skin types, with a focus on secreted proteins. We identified a number of genes that differ dramatically in expression and, among the expressed proteins, neuregulin-1, which is secreted by fibroblasts derived from dark skin, effectively increases the pigmentation of melanocytes in tissue culture and in an artificial skin model and regulates their growth, suggesting that it is one of the major factors determining human skin color.
Formation of extra centrosomal structures is dependent on β-catenin
Authors: Bahmanyar Shirin , Guiney Evan L. , Hatch Emily M. , Nelson W. James , Barth Angela I. M.
Published: August 2010

β-Catenin has important roles in cell–cell adhesion and in the regulation of gene transcription. Mutations that stabilize β-catenin are common in cancer, but it remains unclear how these mutations contribute to cancer progression. β-Catenin is also a centrosomal component involved in centrosome separation. Centrosomes nucleate interphase microtubules and the bipolar mitotic spindle in normal cells, but their organization and function in human cancers are abnormal. Here, we show that expression of stabilized mutant β-catenin, which mimics mutations found in cancer, results in extra non-microtubule nucleating structures that contain a subset of centrosome proteins including γ-tubulin and centrin, but not polo-like kinase 4 (Plk4), SAS-6 or pericentrin. A transcriptionally inactive form of β-catenin also gives rise to abnormal structures of centrosome proteins. HCT116 human colon cancer cell lines, from which the mutant β-catenin allele has been deleted, have reduced numbers of cells with abnormal centrosome structures and S-phase-arrested, amplified centrosomes. RNAi-mediated depletion of β-catenin from centrosomes inhibits S-phase-arrested amplification of centrosomes. These results indicate that β-catenin is required for centrosome amplification, and mutations in β-catenin might contribute to the formation of abnormal centrosomes observed in cancers.
β-Dystroglycan binds caveolin-1 in smooth muscle: a functional role in caveolae distribution and Ca2+ release
Authors: Sharma Pawan , Ghavami Saeid , Stelmack Gerald L. , McNeill Karol D. , Mutawe Mark M. , Klonisch Thomas , Unruh Helmut , Halayko Andrew J.
Published: August 2010

The dystrophin–glycoprotein complex (DGC) links the extracellular matrix and actin cytoskeleton. Caveolae form membrane arrays on smooth muscle cells; we investigated the mechanism for this organization. Caveolin-1 and β-dystroglycan, the core transmembrane DGC subunit, colocalize in airway smooth muscle. Immunoprecipitation revealed the association of caveolin-1 with β-dystroglycan. Disruption of actin filaments disordered caveolae arrays, reduced association of β-dystroglycan and caveolin-1 to lipid rafts, and suppressed the sensitivity and responsiveness of methacholine-induced intracellular Ca2+ release. We generated novel human airway smooth muscle cell lines expressing shRNA to stably silence β-dystroglycan expression. In these myocytes, caveolae arrays were disorganized, caveolae structural proteins caveolin-1 and PTRF/cavin were displaced, the signaling proteins PLCβ1 and Gαq, which are required for receptor-mediated Ca2+ release, were absent from caveolae, and the sensitivity and responsiveness of methacholine-induced intracellular Ca2+ release, was diminished. These data reveal an interaction between caveolin-1 and β-dystroglycan and demonstrate that this association, in concert with anchorage to the actin cytoskeleton, underpins the spatial organization and functional role of caveolae in receptor-mediated Ca2+ release, which is an essential initiator step in smooth muscle contraction.
Spatially defined Dsh–Lgl interaction contributes to directional tissue morphogenesis
Authors: Kaplan Nicole A. , Tolwinski Nicholas S.
Published: August 2010

The process of epithelial morphogenesis defines the structure of epidermal tissue sheets. One such sheet, the ventral epidermis of the Drosophila embryo, shows both intricate segmental patterning and complex cell organization. Within a segment, cells produce hair-like denticles in a stereotypical and highly organized pattern over the surface of the tissue. To understand the cell biological basis of this process, we examined cell shapes and alignments, and looked for molecules that showed an asymmetric distribution in this tissue. We found that apical polarity determinants and adherens junctions were enriched at the dorsal and ventral borders of cells, whereas basolateral determinants were enriched at the anterior and posterior borders. We report that the basolateral determinant Lgl has a novel function in the planar organization of the embryonic epidermis, and this function depends on Dsh and myosin. We conclude that apical–basal proteins, used to establish polarity within a cell, can be independently co-opted to function in epithelial morphogenesis.
SPICE – a previously uncharacterized protein required for centriole duplication and mitotic chromosome congression
Authors: Archinti Marco , Lacasa Cristina , Teixidó-Travesa Neus , Lüders Jens
Published: August 2010

Proper assembly and function of a bipolar mitotic spindle is crucial for faithful bidirectional chromosome segregation during cell division. In animal cells, the two poles of the mitotic spindle are organized by centrosomes, microtubule-organizing structures composed of a pair of centrioles surrounded by the so-called pericentriolar material. Proteomic studies have revealed a large number of centrosome proteins, but many remain uncharacterized. Here, we characterize SPICE, a protein that localizes to spindle microtubules in mitosis and to centrioles throughout the cell cycle. RNAi-mediated depletion of SPICE in human cells impairs centriole duplication and causes severe mitotic defects. SPICE depletion compromises spindle architecture, spindle pole integrity and chromosome congression, even in cells in which centriole duplication has occurred. Our data suggest that SPICE is an important dual-function regulator required for centriole duplication and for proper bipolar spindle formation and chromosome congression in mitosis.
Multisite phosphorylation of Erk5 in mitosis
Authors: DÍaz-RodrÍguez Elena , Pandiella Atanasio
Published: August 2010

The MAP kinase Erk5 plays important roles in cellular proliferation, and has recently been implicated in the regulation of mitosis. The classic pathway of Erk5 activation involves dual phosphorylation at its TEY microdomain by the upstream regulating kinase MEK5. Here we describe a second pathway that controls Erk5 phosphorylation. This pathway is activated in mitotic cells and involves kinase activities distinct from MEK5. Studies aimed at identifying these kinases suggested that CDK1 activity is required to sustain Erk5 phosphorylation in mitosis, as treatment with RO3306, a CDK1 inhibitor, reversed mitotic phosphorylation of Erk5. Moreover, CDK1 co-precipitated with Erk5 in mitotic cells. The mitotic phosphorylation of Erk5 occurs at multiple sites located at its unique C-terminal region, within an Erk5 subdomain that has formerly been implicated in the control of the subcellular location of Erk5. Furthermore, molecular studies indicated that phosphorylation at these sites may participate in the control of the transit of Erk5 between the cytosol and the nucleus, in addition to regulating its transcriptional activity. Together, our results demonstrate the existence of a second Erk5 phosphorylation pathway, that is activated in mitosis, and that may participate in the regulation of Erk5 functions.
Exocytosis, dependent on Ca2+ release from Ca2+ stores, is regulated by Ca2+ microdomains
Authors: Low Jiun T. , Shukla Alka , Behrendorff Natasha , Thorn Peter
Published: August 2010

The relationship between the cellular Ca2+ signal and secretory vesicle fusion (exocytosis) is a key determinant of the regulation of the kinetics and magnitude of the secretory response. Here, we have investigated secretion in cells where the exocytic response is controlled by Ca2+ release from intracellular Ca2+ stores. Using live-cell two-photon microscopy that simultaneously records Ca2+ signals and exocytic responses, we provide evidence that secretion is controlled by changes in Ca2+ concentration [Ca2+] in relatively large-volume microdomains. Our evidence includes: (1) long latencies (>2 seconds) between the rise in [Ca2+] and exocytosis, (2) observation of exocytosis all along the lumen and not clustered around Ca2+ release hot-spots, (3) high affinity (Kd =1.75 μM) Ca2+ dependence of exocytosis, (4) significant reduction in exocytosis in the prescence of cytosolic EGTA, (5) spatial exclusion of secretory granules from the cell membrane by the endoplasmic reticulum, and (6) inability of local Ca2+ responses to trigger exocytosis. These results strongly indicate that the control of exocytosis, triggered by Ca2+ release from stores, is through the regulation of cytosolic [Ca2+] within a microdomain.
Leiomodin-2 is an antagonist of tropomodulin-1 at the pointed end of the thin filaments in cardiac muscle
Authors: Tsukada Takehiro , Pappas Christopher T. , Moroz Natalia , Antin Parker B. , Kostyukova Alla S. , Gregorio Carol C.
Published: August 2010

Regulation of actin filament assembly is essential for efficient contractile activity in striated muscle. Leiomodin is an actin-binding protein and homolog of the pointed-end capping protein, tropomodulin. These proteins are structurally similar, sharing a common domain organization that includes two actin-binding sites. Leiomodin also contains a unique C-terminal extension that has a third actin-binding WH2 domain. Recently, the striated-muscle-specific isoform of leiomodin (Lmod2) was reported to be an actin nucleator in cardiomyocytes. Here, we have identified a function of Lmod2 in the regulation of thin filament lengths. We show that Lmod2 localizes to the pointed ends of thin filaments, where it competes for binding with tropomodulin-1 (Tmod1). Overexpression of Lmod2 results in loss of Tmod1 assembly and elongation of the thin filaments from their pointed ends. The Lmod2 WH2 domain is required for lengthening because its removal results in a molecule that caps the pointed ends similarly to Tmod1. Furthermore, Lmod2 transcripts are first detected in the heart after it has begun to beat, suggesting that the primary function of Lmod2 is to maintain thin filament lengths in the mature heart. Thus, Lmod2 antagonizes the function of Tmod1, and together, these molecules might fine-tune thin filament lengths.
ERK5 is required for VEGF-mediated survival and tubular morphogenesis of primary human microvascular endothelial cells
Authors: Roberts Owain Llŷr , Holmes Katherine , Müller Jürgen , Cross Darren A. E. , Cross Michael J.
Published: August 2010

Extracellular signal-regulated kinase 5 (ERK5) is activated in response to environmental stress and growth factors. Gene ablation of Erk5 in mice is embryonically lethal as a result of disruption of cardiovascular development and vascular integrity. We investigated vascular endothelial growth factor (VEGF)-mediated ERK5 activation in primary human dermal microvascular endothelial cells (HDMECs) undergoing proliferation on a gelatin matrix, and tubular morphogenesis within a collagen gel matrix. VEGF induced sustained ERK5 activation on both matrices. However, manipulation of ERK5 activity by siRNA-mediated gene silencing disrupted tubular morphogenesis without impacting proliferation. Overexpression of constitutively active MEK5 and ERK5 stimulated tubular morphogenesis in the absence of VEGF. Analysis of intracellular signalling revealed that ERK5 regulated AKT phosphorylation. On a collagen gel, ERK5 regulated VEGF-mediated phosphorylation of the pro-apoptotic protein BAD and increased expression of the anti-apoptotic protein BCL2, resulting in decreased caspase-3 activity and apoptosis suppression. Our findings suggest that ERK5 is required for AKT phosphorylation and cell survival and is crucial for endothelial cell differentiation in response to VEGF.
1,5-Anhydroglucitol levels are low irrespective of plasma glucose levels in patients with chronic liver disease
Authors: Koga Masafumi , Murai Jun , Saito Hiroshi , Mukai Mikio , Toya Daishu , Tanaka Nobuyoshi , Kanehara Hideo , Bando Yukihiro , Kasayama Soji
Published: August 2010

Backgound

Serum 1,5-anhydroglucitol (1,5-AG) is a known marker reflecting recent glycaemic control. In this study, we examined serum 1,5-AG levels in chronic liver disease (CLD) patients with and without diabetes mellitus.

Methods

Eighty patients with CLD were compared with 667 subjects without CLD. Glycaemic control of the CLD patients was evaluated by estimated glycated haemoglobin (HbA1C) calculated using the equation by Rohlfing et al. from mean plasma glucose because CLD patients have apparently low HbA1C.

Results

When the study participants were divided into subgroups stratified by HbA1C levels, the CLD patients whose estimated HbA1C levels were less than 7.0% showed significantly lower 1,5-AG than their counterparts of the control subjects. Stepwise multivariable analysis revealed that estimated HbA1C was the significant explanatory variable for 1,5-AG in the CLD patients. However, in the CLD patients with estimated HbA1C less than 5.8%, only hepaplastin test was the significant explanatory variable for 1,5-AG.

Conclusions

Serum 1,5-AG levels are low irrespective of plasma glucose levels in the CLD patients with and without diabetes. The CLD patients who had low serum 1,5-AG levels were associated with deteriorated liver function.
Measurement of lipoprotein particle sizes using dynamic light scattering
Authors: Sakurai Toshihiro , Trirongjitmoah Suchin , Nishibata Yuka , Namita Takeshi , Tsuji Masahiro , Hui Shu-Ping , Jin Shigeki , Shimizu Koichi , Chiba Hitoshi
Published: August 2010

Background

A simple method for the measurement of LDL particle sizes is needed in clinical laboratories because a predominance of small, dense LDL (sd LDL) has been associated with coronary heart disease. We applied dynamic light scattering (DLS) to measure lipoprotein particle sizes, with special reference to sd LDL.

Methods

Human serum lipoproteins isolated by a combination of ultracentrifugation and gel chromatography, or by sequential ultracentrifugation, were measured for particle size using DLS.

Results

The sizes of polystyrene beads, with diameters of 21 and 28 nm according to the manufacturer, were determined by DLS as 19.3 ± 1.0 nm (mean ± SD, n = 11) and 25.5 ± 1.0 nm, respectively. The coefficients of variation for the 21 and 28 nm beads were 5.1% and 3.8% (within-run, n = 11), and 2.9% and 6.2% (between-run, n = 3), respectively. The lipoprotein sizes determined by DLS for lipoprotein fractions isolated by chromatography were consistent with the elution profile. Whole serum, four isolated lipoprotein fractions (CM + VLDL + IDL, large LDL, sd LDL and HDL) and a non-lipoprotein fraction isolated by sequential ultracentrifugation were determined by DLS to be 13.1 ± 7.5, 37.0 ± 5.2, 21.5 ± 0.8, 20.3 ± 1.1, 8.6 ± 1.5 and 8.8 ± 2.0 nm, respectively.

Conclusions

The proposed DLS method can differentiate the sizes of isolated lipoprotein particles, including large LDL and sd LDL, and might be used in clinical laboratories in combination with convenient lipoprotein separation.
Revisiting protocols for burn injury management
Authors: Greenwood John E , Kavanagh Sheila , Mackie Ian P
Published: August 2010

The aim of the study was to reassess surgical protocols developed and introduced in 2005; to examine the changes that have been made and to understand why they have been necessary or felt to be an improvement on the original; and to introduce a new protocol to assist with burn depth assessment. A discussion of how the protocols were developed and the environment in which they are used is undertaken. Four years of experience using the protocols has been accrued. An examination of alterations is performed. The protocols have allowed a single-handed practitioner to deal successfully with a large caseload and enabled a greater understanding of time and resource management. The original development of the protocols was worthwhile but their continued evolution by frequent audit and evaluation of innovations in burn care is mandatory.
An overview on the history and concept of care pathways as complex interventions
Authors: Vanhaecht Kris , Panella Massimiliano , van Zelm Ruben , Sermeus Walter
Published: August 2010

Care pathways, also known as clinical pathways, critical pathways or integrated care pathways, are used all over the world. Although they are used internationally, there are still a large number of misunderstandings. The goal of this paper is to provide an overview on the history of pathways and how pathways are actually perceived and defined. Pathways are more than just a document in the patient record. They are a concept for making patient-focused care operational and supporting the modelling of patient groups with different levels of predictability. Pathways are a method within the field of continuous quality improvement and are used in daily practice as a product in the patient record. This paper explains these different issues and provides an extensive list of references that should support pathway facilitators, clinicians, managers and policy-makers in their search for excellence.
Care pathways and patient safety: key concepts, patient outcomes and related interventions
Authors: Dy Sydney , Gurses Ayse P
Published: August 2010

Although care pathways often target efficiency of care through mapping and standardizing care processes, care can also be improved by reducing patient safety events, such as complications. In this paper, the authors review key concepts and literature relevant to parallels between patient safety and pathway interventions, as well as patient safety issues that should be considered in pathway development and implementation. Both care pathways and patient safety interventions are more likely to be effective when based on a theoretical framework related to human or systems factors or behaviour. Care pathways can target patient safety outcomes, but can also produce new hazards, through applying standards too broadly, reducing adaptability to complex situations or changing care processes in unforeseen ways. Both pathways and safety interventions must also be efficient and consider the opportunity costs of the time needed for providers to implement the intervention. Further research should explore how best to standardize care when needed, while evaluating how best to prevent and monitor hazards, allow for innovation and adaptability to customize care when appropriate, and continue to develop new methods for improving quality.
Competencies for managers of care pathways: an exploratory study
Authors: Verkerk-Geelhoed Jeannette , van Zelm Ruben
Published: August 2010

This study was conducted to prioritize the competencies for the operational manager regarding the management of care pathways. In the Schouten & Nelissen Competence Model (SNCM), the operational manager works according to the management style ‘task-centred leader/producer’. A measurement instrument was developed to conduct a Delphi study. The competencies on which complete consensus has been reached were presented to a focus group in the second phase of the study. The Delphi was carried out with five experts in three rounds. Complete consensus on 30 competencies was reached. The focus group consisted of three participants and produced 10 core competencies. These belong to the level of operational management and the management style ‘task-centred leader/producer’ in the SNCM. The number of experts in both the Delphi study and focus group was limited, which may have influenced the outcome. There is hardly any literature on competencies operational managers' need for managing care pathways. This study has produced 10 competencies that the operational manager needs for coordinating care pathways. These core competencies can be used to specify or complete the competence profile of operational managers working with care pathways.
An updated and simplified method for bile duct cannulation of rats
Authors: Tønsberg H , Holm R , Bjerregaard T G , Boll J B , Jacobsen J , Müllertz A
Published: August 2010

The single bile duct cannulated rat model was used for short-term intestinal absorption studies. Rats were randomly assigned to three groups: (A) bile duct cannulated, (B) control laparotomy and (C) no treatment. The body weight and health of the rats were monitored before and until day 5 after surgery, while bile flow was measured in group A on day 2. On the fifth day, tail vein blood was harvested, and alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and total bilirubin were quantified. Analysis of variance showed no significant difference between the groups for these parameters. This study demonstrated the suitability of a surgical rat model feasible for evaluation of the impact of bile in pharmacokinetic studies.
Differential assembly of 16S rRNA domains during 30S subunit formation
Authors: Xu Zhili , Culver Gloria M.
Published: August 2010

Rapid and accurate assembly of the ribosomal subunits, which are responsible for protein synthesis, is required to sustain cell growth. Our best understanding of the interaction of 30S ribosomal subunit components (16S ribosomal RNA [rRNA] and 20 ribosomal proteins [r-proteins]) comes from in vitro work using Escherichia coli ribosomal components. However, detailed information regarding the essential elements involved in the assembly of 30S subunits still remains elusive. Here, we defined a set of rRNA nucleotides that are critical for the assembly of the small ribosomal subunit in E. coli. Using an RNA modification interference approach, we identified 54 nucleotides in 16S rRNA whose modification prevents the formation of a functional small ribosomal subunit. The majority of these nucleotides are located in the head and interdomain junction of the 30S subunit, suggesting that these regions are critical for small subunit assembly. In vivo analysis of specific identified sites, using engineered mutations in 16S rRNA, revealed defective protein synthesis capability, aberrant polysome profiles, and abnormal 16S rRNA processing, indicating the importance of these residues in vivo. These studies reveal that specific segments of 16S rRNA are more critical for small subunit assembly than others, and suggest a hierarchy of importance.
Medication History Reconciliation by Clinical Pharmacists in Elderly Inpatients Admitted from Home or a Nursing Home
Authors: Steurbaut Stephane , Leemans Lies , Leysen Tinne , De Baere Eva , Cornu Pieter , Mets Tony , Dupont Alain G
Published: August 2010

Background:

Accurate medication histories at hospital admission are an important element of medication safety. Discrepancies may have clinically significant consequences, especially in the elderly population.

Objective:

To assess the clinical pharmacist's performance in obtaining patients' medication histories and in reconciling these data with the medical records and medication orders and whether the patients' residential situation prior to hospitalization influences the number of drug discrepancies.

Methods:

A prospective observational study was conducted at a 29-bed acute geriatric ward of a Belgian university hospital. Medication histories acquired by clinical pharmacists were compared with those documented in the medical records by the attending physicians. All discrepancies were identified and categorized by an independent pharmacist and were scored for their clinical relevance in consensus by a senior internist and a senior geriatrician.

Results:

Of the 215 screened geriatric (aged ≥66 years) patients admitted between October 27, 2007, and September 23, 2008, 197 were enrolled in the study. For patients living in the community, as well as those residing in a nursing home prior to hospitalization, clinical pharmacists identified significantly more preadmission drugs compared with physicians, with a median number of 8 correctly identified medications versus 6, respectively (p < 0.001). Extra identified drugs consisted of over-the-counter as well as prescription medications. Furthermore, 117 other medication discrepancies were noted, mainly related to erroneous drug identification and incorrect drug dose. In all, the clinical pharmacists identified 379 (24.2%) medication discrepancies, of which 188 (49.6%) were judged clinically relevant.

Conclusions:

Pharmacist-acquired medication histories enhance the medication reconciliation process, both in patients residing at home and in a nursing home prior to hospitalization. A focus should be placed on seamless care procedures that facilitate the transfer of medication histories between primary and secondary care in both of these populations.
High-Dose Ciprofloxacin for Serious Gram-Negative Infection in an Obese, Critically III Patient Receiving Continuous Venovenous Hemodiafiltration
Authors: Utrup Theresa R , Mueller Eric W , Healy Daniel P , Calicut Rachael A , Peterson John D , Hurford William E
Published: August 2010

Objective:

To describe the pharmacokinetic profile and clinical outcome associated with high-dose ciprofloxacin therapy in a patient with the triad of extreme obesity, multiple organ failure, and deep-seated infection.

Case Summary:

A 45-year-old, class 3 obese (185 kg; body mass index 53.7), critically ill trauma patient receiving continuous venovenous hemodiafiltration (CVVHOF) was treated with ciprofloxacin 800 mg intravenously every 12 hours for presumed Enterobacter aerogenes (ciprofloxacin minimum inhibitory concentration [MIC] ≤1 μg/mL) lumbar spine osteomyelitis. Four sequential plasma ciprofloxacin samples were obtained and analyzed to determine the steady-state pharmacokinetic profile. The observed steady-state maximum (Cmax) and calculated minimum (Cmin) ciprofloxacin plasma concentrations measured on treatment day 8 were 13 μg/mL and 4.8 μg/mL, respectively, corresponding to an estimated half-life, area under the curve (AUC0-24), total systemic clearance, and clearance by CVVHDF of 7.6 hours, 132 μg·h/mL, 139 mL/min, and 26 mL/min, respectively. These concentrations produced AUC0-24/MIC ratios >125 and plasma Cmax/MIC ratios >10 for MICs ≤1 μg/mL. Intravenous colistin and polymyxin B lumbar wound irrigation were initiated on ciprofloxacin days 12 and 15, respectively, for concomitant multidrug-resistant Acinetobacter baumannii infection. Lumbar tissue cultures on day 24 of ciprofloxacin therapy demonstrated no growth, coinciding with overall improvement of the invasive wound, A week later, the patient developed worsening septic shock and died secondary to an occult subdiaphragmatic abscess.

Discussion:

Pharmacodynamic outcome studies suggest that AUC0-24/MIC ratios >125 and plasma Cmax/MIC ratios >10 are good predictors of clinical and microbiologic success of ciprofloxacin against gram-negative pathogens. These pharmacodynamic goals were achieved in the plasma with high-dose ciprofloxacin for MICs ≤1 μg/mL.

Conclusions:

Critically ill obese patients with deep-seated infection involving organisms with MICs >0.5 μg/mL likely require ciprofloxacin dosages greater than traditional daily doses of 400–800 mg during CVVHDF to achieve optimal pharmacodynamic targets.