bluuebunny/crossref_metadata_2025_split huggingface dataset:
Smooth muscle signalling pathways in health and disease
Authors: Kim H. R. , Appel S. , Vetterkind S. , Gangopadhyay S. S. , Morgan K. G. 
Published: October 2008

IntroductionMechanisms that regulate LC20 phosphorylationMechanisms that regulate the access of myosin to actinMechanisms that regulate cytoskeletal remodellingConclusionsAbstractSmooth muscle contractile activity is a major regulator of function of the vascular system, respiratory system, gastrointestinal system and the genitourinary systems. Malfunction of contractility in these systems leads to a host of clinical disorders, and yet, we still have major gaps in our understanding of the molecular mechanisms by which contractility of the differentiated smooth muscle cell is regulated. This review will summarize recent advances in the molecular understanding of the regulation of smooth muscle myosin activityviaphosphorylation/dephosphorylation of myosin, the regulation of the accessibility of actin to myosinvia theactin‐binding proteins calponin and caldesmon, and the remodelling of the actin cytoskeleton. Understanding of the molecular ‘players’ should identify target molecules that could point the way to novel drug discovery programs for the treatment of smooth muscle disorders such as cardiovascular disease, asthma, functional bowel disease and pre‐term labour.
Junior doctor strike model of care: Reduced access block and predominant Fellow of the Australasian College for Emergency Medicine staffing improve emergency department performance
Authors: Thornton Vanessa , Hazell Wayne 
Published: October 2008

AbstractObjective:  To describe the response and analyse ED performance during a 5‐day junior doctor strike.Methods:  Data were collected via the patient information management computer system. Key performance indicators included percentage seen within maximum waiting times per triage category (TC), ED length of stay, emergency medicine patients who did not wait to be seen, hospital bed occupancy and access block percentage. Comparisons were made for the same 5 days before the strike (BS), during the strike (S) and after the strike.Results:  Total doctor's shifts BS were 78.66 with 25% of these shifts being Fellow of the Australasian College for Emergency Medicine (FACEM) shifts. FACEM shifts were more common during the S period at 75% (P < 0.001). Total attendances (BS 631 vs S 596, P = 0.22) and TC percentages (P‐values for TC 1, 2, 3, 4, 5, respectively, 1.0, 0.55, 0.88, 0.97, 0.46) in the BS, S and after‐the‐strike periods were not significantly different. Despite fewer total doctor shifts, the FACEM predominant model of care during the strike resulted in better percentages seen within the maximum waiting times for TC3 (66%), TC4 (78%) and TC5 (86%) (all P < 0.001). There was a reduction in patients who did not wait to be seen (28 BS vs 5 S, P < 0.001), ED length of stay (admissions: BS 451 min vs S 258 min, P < 0.001; discharges: BS 233 min vs S 144 min, P < 0.02) and referrals to inpatient services (P = 0.02). This occurred with reduced bed point occupancy of 66% and a consequent reduction in access block.Conclusion:  FACEM staffing and reduced access block were significant factors in improved ED performance.
Perceived Barriers and Benefits to Research Participation Among School Administrators*
Authors: Befort Christie , Lynch Rachel , James Rochelle L. , Carroll Shawna L. , Nollen Nicole , Davis Ann 
Published: October 2008

ABSTRACTBackground:  Gaining access to conduct research in schools can be challenging, but little has been done to directly assess school administrators’ perceptions of research. The purpose of this qualitative study was to increase understanding of barriers and benefits of research participation as perceived by superintendents and principals in elementary, middle, and high school settings.Methods:  Administrators (14 elementary, 14 middle, and 15 high school principals and 14 superintendents; total n = 57) were randomly selected from across a Midwestern state and were interviewed by phone following a semistructured guide of questions.Results:  Six major themes were consistent across principals and superintendents and reached saturation. Themes indicated that administrators were interested in research projects that (1) provide tangible benefits to their school, (2) are consistent with their academic mission, (3) are not burdensome, (4) do not take place during state assessment time or other busy times, and (5) are credible and noncontroversial. Previous research experiences leave a lasting impression on administrators and influenced their future decisions.Conclusions:  Findings indicate that researchers should carefully frame their research in terms of how it will directly benefit the school, keeping in mind that academic performance is the top priority for school administrators. Researchers should be very clear and realistic about the time commitments, how the research results will be provided, and how the study may be used to improve the school’s academic mission.
Directed evolution of Aspergillus niger glucoamylase to increase thermostability
Authors: McDaniel Allison , Fuchs Erica , Liu Ying , Ford Clark 
Published: October 2008

SummaryUsing directed evolution and site‐directed mutagenesis, we have isolated a highly thermostable variant of Aspergillus niger glucoamylase (GA), designated CR2‐1. CR2‐1 includes the previously described mutations Asn20Cys and Ala27Cys (forming a new disulfide bond), Ser30Pro, Thr62Ala, Ser119Pro, Gly137Ala, Thr290Ala, His391Tyr and Ser436Pro. In addition, CR2‐1 includes several new putative thermostable mutations, Val59Ala, Val88Ile, Ser211Pro, Asp293Ala, Thr390Ser, Tyr402Phe and Glu408Lys, identified by directed evolution. CR2‐1 GA has a catalytic efficiency (kcat/Km) at 35°C and a specific activity at 50°C similar to that of wild‐type GA. Irreversible inactivation tests indicated that CR2‐1 increases the free energy of thermoinactivation at 80°C by 10 kJ mol−1 compared with that of wild‐type GA. Thus, CR2‐1 is more thermostable (by 5 kJ mol−1 at 80°C) than the most thermostable A. niger GA variant previously described, THS8. In addition, Val59Ala and Glu408Lys were shown to individually increase the thermostability in GA variants by 1 and 2 kJ mol−1, respectively, at 80°C.
Shift of temporal responsivity curve of pattern reversal visually evoked potentials to lower frequencies in patients after recovery from optic neuritis
Authors: Okuno Takashi , Oku Hidehiro , Sugasawa Jun , Utsumi Takashi , Ikeda Tsunehiko 
Published: October 2008

Abstract.Purpose:  To investigate the responsivity of the visual system to pattern reversal stimuli of different reversal rates in optic neuritis patients after a full recovery of visual acuity.Methods:  Thirty eyes with optic neuritis and 30 eyes of age‐matched normal controls were studied. Steady‐state pattern reversal visually evoked potentials (P‐VEPs) were recorded using reversal rates of 2, 10, 20 and 30 Hz. The relationship between the amplitudes of the P‐VEPs and reversal rate was analysed. The temporal responsivity curves of the two groups were compared.Results:  The means ± standard deviations (SDs) of the P‐VEP amplitudes in the normal controls were 7.08 ± 2.97, 8.60 ± 2.37, 4.69 ± 1.99 and 2.11 ± 1.24 μV at reversal rates of 2, 10, 20 and 30 Hz, respectively. The comparable amplitudes from the recovered optic neuritis patients were 8.01 ± 2.45, 6.57 ± 2.54, 2.73 ± 1.76 and 0.94 ± 1.27 μV. The mean amplitude of the P‐VEPs elicited at a reversal rate of 2 Hz was not significantly different from that of normal controls. However, the amplitudes of the P‐VEPs at reversal rates of 10, 20 and 30 Hz were significantly smaller in the recovered optic neuritis patients than in the controls. In addition, the average maximum amplitude of the P‐VEPs was obtained at 10 Hz in normal subjects and at 2 Hz in the recovered optic neuritis patients.Conclusion:  These results indicate that the peak of the temporal tuning curve of the P‐VEP shifts to lower frequencies in recovered optic neuritis eyes, and that the reduced amplitudes at higher frequencies seem to be characteristic of the visual system of patients with recovered optic neuritis.
Observations of avoidance reactions of migrating salmon to a mobile survey vessel in a riverine environment
Authors: Xie Yunbo , Michielsens Catherine G.J. , Gray Andrew P. , Martens Fiona J. , Boffey Jacqueline L. 
Published: October 2008

Detailed avoidance reactions of adult migrating salmon to a mobile survey vessel were successfully observed with side-looking dual-frequency identification sonar (DIDSON) in the lower Fraser River (British Columbia, Canada). Both adult sockeye ( Oncorhynchus nerka ) and pink salmon ( Oncorhynchus gorbuscha ) returning to the river were found to avoid the approaching vessel by initiating lateral movements away from the vessel, making the fish unlikely to be insonified by the downward-looking transducers towed by the vessel. The vessel was found to have an estimated mean interference range of 4 m from its propeller. Analyses of the data concluded that once the vessel and fish were separated by more than 7 m, the vessel no longer affected the normal migration behaviour of the fish.
Comparison of three microquantity techniques for measuring total lipids in fish
Authors: Lu Yuehan , Ludsin Stuart A. , Fanslow David L. , Pothoven Steven A. 
Published: October 2008

To measure lipids in juvenile and adult fishes, we refined three microquantity approaches (microgravimetric, microcolorimetric sulfophosphovanillan (SPV), and Iatroscan thin layer chromatography – flame ionization detection (TLC–FID)) that were originally developed to measure lipids in small aquatic invertebrates. We also evaluated their precision and comparability by quantifying the total lipid content of age-1+ walleye ( Sander vitreus ), yellow perch ( Perca flavescens ), and lake whitefish ( Coregonus clupeaformis ) collected in Lake Erie (US–Canada), Lake Michigan (USA), and Muskegon Lake (Michigan, USA). Our findings demonstrate that (i) microquantity approaches provide estimates of total lipids in juvenile and adult fishes similar to those of more traditional macroquantity approaches, (ii) the microcolorimetric SPV and microgravimetric approaches produce near identical estimates of total lipid content, and (iii) the Iatroscan TLC–FID approach underestimates total lipids relative to the other approaches for individuals with high lipid levels. Ultimately, our research makes available additional techniques for measuring total lipid content of fishes that are less expensive than traditional techniques, owing to a reduced need for large quantities of samples and solvents.
Src and ADAM-17–Mediated Shedding of Transforming Growth Factor-α Is a Mechanism of Acute Resistance to TRAIL
Authors: Van Schaeybroeck Sandra , Kelly Donal M. , Kyula Joan , Stokesberry Susan , Fennell Dean A. , Johnston Patrick G. , Longley Daniel B. 
Published: October 2008

Abstract
Tumor necrosis factor–related apoptosis-inducing ligand (TRAIL/Apo-2L) has emerged as a promising anticancer agent. However, resistance to TRAIL is likely to be a major problem, and sensitization of cancer cells to TRAIL may therefore be an important anticancer strategy. In this study, we examined the effect of the epidermal growth factor receptor (EGFR)–tyrosine kinase inhibitor (TKI) gefitinib and a human epidermal receptor 2 (HER2)–TKI (M578440) on the sensitivity of human colorectal cancer (CRC) cell lines to recombinant human TRAIL (rhTRAIL). A synergistic interaction between rhTRAIL and gefitinib and rhTRAIL and M578440 was observed in both rhTRAIL-sensitive and resistant CRC cells. This synergy correlated with an increase in EGFR and HER2 activation after rhTRAIL treatment. Furthermore, treatment of CRC cells with rhTRAIL resulted in activation of the Src family kinases (SFK). Importantly, we found that rhTRAIL treatment induced shedding of transforming growth factor-α (TGF-α) that was dependent on SFK activity and the protease ADAM-17. Moreover, this shedding of TGF-α was critical for rhTRAIL-induced activation of EGFR. In support of this, SFK inhibitors and small interfering RNAs targeting ADAM-17 and TGF-α also sensitized CRC cells to rhTRAIL-mediated apoptosis. Taken together, our findings indicate that both rhTRAIL-sensitive and resistant CRC cells respond to rhTRAIL treatment by activating an EGFR/HER2-mediated survival response and that these cells can be sensitized to rhTRAIL using EGFR/HER2-targeted therapies. Furthermore, this acute response to rhTRAIL is regulated by SFK-mediated and ADAM-17–mediated shedding of TGF-α, such that targeting SFKs or inhibiting ADAM-17, in combination with rhTRAIL, may enhance the response of CRC tumors to rhTRAIL. [Cancer Res 2008;68(20):8312–21]
Phosphorylated Caveolin-1 Regulates Rho/ROCK-Dependent Focal Adhesion Dynamics and Tumor Cell Migration and Invasion
Authors: Joshi Bharat , Strugnell Scott S. , Goetz Jacky G. , Kojic Liliana D. , Cox Michael E. , Griffith Obi L. , Chan Simon K. , Jones Steven J. , Leung Sher-Ping , Masoudi Hamid , Leung Samuel , Wiseman Sam M. , Nabi Ivan R. 
Published: October 2008

Abstract
Rho/ROCK signaling and caveolin-1 (Cav1) are implicated in tumor cell migration and metastasis; however, the underlying molecular mechanisms remain poorly defined. Cav1 was found here to be an independent predictor of decreased survival in breast and rectal cancer and significantly associated with the presence of distant metastasis for colon cancer patients. Rho/ROCK signaling promotes tumor cell migration by regulating focal adhesion (FA) dynamics through tyrosine (Y14) phosphorylation of Cav1. Phosphorylated Cav1 is localized to protrusive domains of tumor cells and Cav1 tyrosine phosphorylation is dependent on Src kinase and Rho/ROCK signaling. Increased levels of phosphorylated Cav1 were associated with elevated GTP-RhoA levels in metastatic tumor cells of various tissue origins. Stable expression and knockdown studies of Cav1 in tumor cells showed that phosphorylated Cav1 expression stimulates Rho activation, stabilizes FAK association with FAs, and promotes cell migration and invasion in a ROCK-dependent and Src-dependent manner. Tyrosine-phosphorylated Cav1, therefore, functions as an effector of Rho/ROCK signaling in the regulation of FA turnover and, thereby, tumor cell migration and invasion. These studies define a feedback loop between Rho/ROCK, Src, and phosphorylated Cav1 in tumor cell protrusions, identifying a novel function for Cav1 in tumor metastasis that may contribute to the poor prognosis of some Cav1-expressing tumors. [Cancer Res 2008;68(20):8210–20]
Spinal Glioma: Platelet-Derived Growth Factor B–Mediated Oncogenesis in the Spinal Cord
Authors: Hitoshi Yasuyuki , Harris Brent T. , Liu Huan , Popko Brian , Israel Mark A. 
Published: October 2008

Abstract
Human platelet-derived growth factor B (hPDGFB) has been characterized in vitro and shown to mediate numerous cellular responses including glial proliferation and differentiation. Expression of PDGFB is thought to be important in the pathogenesis of glioma and several animal models of cerebral glioma based on PDGF expression have been described. To examine whether PDGF could contribute to the pathogenesis of spinal cord glioma, we developed transgenic mice that express hPDGFB under the control of a tetracycline-responsive element (TRE/hPDGFB). These TRE/hPDGFB mice were mated with transgenic mice expressing the tetracycline transcriptional activator (tet-off), tTA, regulated by the human glial fibrillary acidic protein (GFAP) promoter and exhibiting uniquely strong promoter activity in the spinal cord. These transgenic mice (GFAP/tTA:TRE/hPDGFB) expressed hPDGFB in GFAP-expressing glia in a manner responsive to doxycycline administration. Without doxycycline, almost all GFAP/tTA:TRE/hPDGFB mice developed spinal cord neoplasms resembling human mixed oligoastrocytoma. Tumorigenesis in these animals was suppressed by doxycycline. To further examine the importance of PDGFB in mouse primary intramedullary spinal cord tumors, we also created transgenic mice expressing hPDGFB under the control of the human GFAP promoter (GFAP/hPDGFB). These GFAP/hPDGFB mice also developed spinal oligoastrocytoma. PDGFB can mediate the development of mouse spinal tumors that are histologically and pathologically indistinguishable from primary intramedullary spinal tumors of humans and may provide opportunities for both novel insights into the pathogenesis of these tumors and the development of new therapeutics. [Cancer Res 2008;68(20):8507–15]
Pleiotropic Biological Activities of Alternatively Spliced TMPRSS2/ERG Fusion Gene Transcripts
Authors: Wang Jianghua , Cai Yi , Yu Wendong , Ren Chengxi , Spencer David M. , Ittmann Michael 
Published: October 2008

Abstract
TMPRSS2/ERG gene fusions are found in the majority of prostate cancers; however, there is significant heterogeneity in the 5′ region of the alternatively spliced fusion gene transcripts. We have found that there is also significant heterogeneity within the coding exons as well. There is variable inclusion of a 72-bp exon and other novel alternatively spliced isoforms. To assess the biological significance of these alternatively spliced transcripts, we expressed various transcripts in primary prostatic epithelial cells (PrEC) and in an immortalized PrEC line, PNT1a. The fusion gene transcripts promoted proliferation, invasion, and motility with variable activities that depended on the structure of the 5′ region encoding the TMPRSS2/ERG fusion and the presence of the 72-bp exon. Cotransfection of different isoforms further enhanced biological activity, mimicking the situation in vivo, in which multiple isoforms are expressed. Finally, knockdown of the fusion gene in VCaP cells resulted in inhibition of proliferation in vitro and tumor progression in an in vivo orthotopic mice model. Our results indicate that TMPRSS2/ERG fusion isoforms have variable biological activities promoting tumor initiation and progression and are consistent with our previous clinical observations indicating that certain TMPRSS2/ERG fusion isoforms are significantly correlated with more aggressive disease. [Cancer Res 2008;68(20):8516–24]
ATP Citrate Lyase: Activation and Therapeutic Implications in Non–Small Cell Lung Cancer
Authors: Migita Toshiro , Narita Tadahito , Nomura Kimie , Miyagi Erika , Inazuka Fumika , Matsuura Masaaki , Ushijima Masaru , Mashima Tetsuo , Seimiya Hiroyuki , Satoh Yukitoshi , Okumura Sakae , Nakagawa Ken , Ishikawa Yuichi 
Published: October 2008

Abstract
Enhanced glucose and lipid metabolism is one of the most common properties of malignant cells. ATP citrate lyase (ACLY) is a key enzyme of de novo fatty acid synthesis responsible for generating cytosolic acetyl-CoA and oxaloacetate. To evaluate its role in lung cancer progression, we here analyzed ACLY expression in a subset of human lung adenocarcinoma cell lines and showed a relationship with the phosphatidyl-inositol-3 kinase–Akt pathway. The introduction of constitutively active Akt into cells enhanced the phosphorylation of ACLY, whereas dominant-negative Akt caused attenuation. In human lung adenocarcinoma samples, ACLY activity was found to be significantly higher than in normal lung tissue. Immunohistochemical analysis further showed phosphorylated ACLY overexpression in 162 tumors, well-correlating with stage, differentiation grade, and a poorer prognosis. Finally, to show the therapeutic potential and mechanism of ACLY inhibition for lung cancer treatment, we assessed the effect of RNA interference targeting ACLY on lipogenesis and cell proliferation in A549 cells. ACLY inhibition resulted in growth arrest in vitro and in vivo. Interestingly, increased intracellular lipids were found in ACLY knockdown cells, whereas de novo lipogenesis was inhibited. Supplementation of insulin could rescue the proliferative arrest elicited by ACLY inhibition; however, in contrast, fatty acid palmitate induced cell death. Taken together, these findings suggest that ACLY is involved in lung cancer pathogenesis associated with metabolic abnormality and might offer a novel therapeutic target. [Cancer Res 2008;68(20):8547–54]
Early Therapy Evaluation of Combined Anti–Death Receptor 5 Antibody and Gemcitabine in Orthotopic Pancreatic Tumor Xenografts by Diffusion-Weighted Magnetic Resonance Imaging
Authors: Kim Hyunki , Morgan Desiree E. , Buchsbaum Donald J. , Zeng Huadong , Grizzle William E. , Warram Jason M. , Stockard Cecil R. , McNally Lacey R. , Long Joshua W. , Sellers Jeffrey C. , Forero Andres , Zinn Kurt R. 
Published: October 2008

Abstract
Early therapeutic efficacy of anti–death receptor 5 antibody (TRA-8) combined with gemcitabine was measured using diffusion-weighted magnetic resonance imaging (DWI) in an orthotopic pancreatic tumor model. Groups 1 to 4 of severe combined immunodeficient mice (n = 5–7 per group) bearing orthotopically implanted, luciferase-positive human pancreatic tumors (MIA PaCa-2) were subsequently (4–5 weeks thereafter) injected with saline (control), gemcitabine (120 mg/kg), TRA-8 (200 μg), or TRA-8 combined with gemcitabine, respectively, on day 0. DWI, anatomic magnetic resonance imaging, and bioluminescence imaging were done on days 0, 1, 2, and 3 after treatment. Three tumors from each group were collected randomly on day 3 after imaging, and terminal deoxyribonucleotidyl transferase–mediated dUTP nick end labeling staining was done to quantify apoptotic cellularity. At just 1 day after starting therapy, the changes of apparent diffusion coefficient (ADC) in tumor regions for group 3 (TRA-8) and group 4 (TRA-8/Gem) were 21 ± 9% (mean ± SE) and 27 ± 3%, respectively, significantly higher (P &lt; 0.05) than those of group 1 (−1 ± 5%) and group 2 (−2 ± 4%). There was no statistical difference in tumor volumes for the groups at this time. The mean ADC values of groups 2 to 4 gradually increased over 3 days, which were concurrent with tumor volume regressions and bioluminescence signal decreases. Apoptotic cell densities of tumors in groups 1 to 4 were 0.7 ± 0.4%, 0.6 ± 0.2%, 3.1 ± 0.9%, and 4.7 ± 1.0%, respectively, linearly proportional to the ADC changes on day 1. Further, the ADC changes were highly correlated with the previously reported mean survival times of animals treated with the same agents and doses. This study supports the clinical use of DWI for pancreatic tumor patients for early assessment of drug efficacy. [Cancer Res 2008;68(20):8369–76]