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Find video protocols related to scientific articles indexed in Pubmed.
'Catching chlamydia': combining cash incentives and community pharmacy access for increased chlamydia screening, the view of young people.
Aust J Prim Health
PUBLISHED: 04-10-2015
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In Australia and elsewhere, chlamydia screening rates for those aged between 16 and 30 years continue to be low. Innovative, age-appropriate approaches are necessary to increase chlamydia screening among this target group to prevent short- and long-term consequences of the infection such as pelvic inflammatory disease, chronic pelvic pain, ectopic pregnancy and infertility. Studies have demonstrated that offering chlamydia screening in community pharmacies may be a useful adjunct to current screening services. Approximately 90% of Australians visit a pharmacy at least once a year. Chlamydia screening and education in community pharmacies with remuneration may provide another option for opportunistic testing as part of a national chlamydia screening scheme. Compensation is an accepted practice in the field of research and has been demonstrated to improve adherence to health promotion activities. In 2011, a cross-sectional study of community pharmacy-based chlamydia screening offered in conjunction with an A$10 cash incentive to participate was conducted in the Australian Capital Territory. As part of this study young people were asked about their experience of, and views about, pharmacy-based chlamydia screening. The views of consented participants were collected using the one-page questionnaire consisting of 10 closed questions and one open-ended question. Participants completed the questionnaire when they returned their urine sample and before being given the cash incentive. Overall participants were highly satisfied with the pharmacy-based chlamydia screening service. Over 60% of questionnaire respondents felt that the payment did affect their decision to have the chlamydia test, and 23% stated that it made no difference. Young people reported that pharmacy-based screening is acceptable and convenient. Accessibility and the small cash incentive played significant roles in increasing participation.
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Sleep Onset and Night Waking Insomnias in Preschoolers with Psychiatric Disorders.
Child Psychiatry Hum Dev
PUBLISHED: 10-08-2014
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This study examined the nature and prevalence of diagnostically defined sleep disorders, including Sleep Onset Insomnia (SOI) and Night Waking Insomnia (NWI), in a sample of 183 young children admitted to an early childhood psychiatric day treatment program. A semi-structured diagnostic interview, the Diagnostic Infant and Preschool Assessment, was used to assess for sleep and other psychiatric disorders. Daily sleep diaries and the Child Behavior Checklist were also examined. 41 % of children met criteria for a sleep disorder; 23 % met diagnostic criteria for SOI and 4 % met criteria for NWI, with an additional 14 % meeting criteria for both (SOI + NWI). Sleep-disordered children demonstrated longer latency to sleep onset, longer and more frequent night awakenings, less total sleep, and lower sleep efficiency than non-sleep disordered participants. Diagnosable sleep disorders, particularly SOI, were quite common in this acute clinical sample, exceeding previous estimates obtained in community and pediatric practice samples.
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Healthy Amistad: improving the health of people with severe mental illness.
Issues Ment Health Nurs
PUBLISHED: 09-27-2014
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Here, we report evaluation results of implementing a health promotion program for individuals with serious mental illnesses. Healthy Amistad aimed to address four behaviors: physical inactivity, nutrition choices, smoking, and seeking access to health care. The evaluation employed a mixed-method study design to assess changes in the health of individuals in the program. Process measures assessed the implementation of the program. A pre-post examination was used to compare data associated with behaviors. Data sources included the 2008 and 2009 annual surveys, clinical data, interviews for staff, interviews with members, and an on-site observation. Participants were staff and members of Amistad. Those involved with the Peer Patient Navigator lost weight; new physically active activities were being offered. A new salad bar and healthier menu was offered in the Amistad cafeteria. Interviews revealed that 11 members lost a total of 150 pounds. The percentage reporting visits to an emergency room more than once in the last 6 months decreased from 58% to 37%, the percentage calling the crisis line less often increased from 75% to 86%, and the percentage reporting that they had become more satisfied with their life since joining Amistad improved from 76% to 88%. Individuals with serious mental illnesses are benefiting from programs that focus on the mitigation of disease states manifested from issues with physical inactivity, nutrition, smoking, and health access. Evaluation of the Healthy Amistad program has shown a positive influence.
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Common variants near ABCA1, AFAP1 and GMDS confer risk of primary open-angle glaucoma.
Nat. Genet.
PUBLISHED: 08-31-2014
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Primary open-angle glaucoma (POAG) is a major cause of irreversible blindness worldwide. We performed a genome-wide association study in an Australian discovery cohort comprising 1,155 cases with advanced POAG and 1,992 controls. We investigated the association of the top SNPs from the discovery stage in two Australian replication cohorts (932 cases and 6,862 controls total) and two US replication cohorts (2,616 cases and 2,634 controls total). Meta-analysis of all cohorts identified three loci newly associated with development of POAG. These loci are located upstream of ABCA1 (rs2472493[G], odds ratio (OR) = 1.31, P = 2.1 × 10(-19)), within AFAP1 (rs4619890[G], OR = 1.20, P = 7.0 × 10(-10)) and within GMDS (rs11969985[G], OR = 1.31, P = 7.7 × 10(-10)). Using RT-PCR and immunolabeling, we show that these genes are expressed within human retina, optic nerve and trabecular meshwork and that ABCA1 and AFAP1 are also expressed in retinal ganglion cells.
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Discovery and optimization of small-molecule ligands for the CBP/p300 bromodomains.
J. Am. Chem. Soc.
PUBLISHED: 06-19-2014
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Small-molecule inhibitors that target bromodomains outside of the bromodomain and extra-terminal (BET) sub-family are lacking. Here, we describe highly potent and selective ligands for the bromodomain module of the human lysine acetyl transferase CBP/p300, developed from a series of 5-isoxazolyl-benzimidazoles. Our starting point was a fragment hit, which was optimized into a more potent and selective lead using parallel synthesis employing Suzuki couplings, benzimidazole-forming reactions, and reductive aminations. The selectivity of the lead compound against other bromodomain family members was investigated using a thermal stability assay, which revealed some inhibition of the structurally related BET family members. To address the BET selectivity issue, X-ray crystal structures of the lead compound bound to the CREB binding protein (CBP) and the first bromodomain of BRD4 (BRD4(1)) were used to guide the design of more selective compounds. The crystal structures obtained revealed two distinct binding modes. By varying the aryl substitution pattern and developing conformationally constrained analogues, selectivity for CBP over BRD4(1) was increased. The optimized compound is highly potent (Kd = 21 nM) and selective, displaying 40-fold selectivity over BRD4(1). Cellular activity was demonstrated using fluorescence recovery after photo-bleaching (FRAP) and a p53 reporter assay. The optimized compounds are cell-active and have nanomolar affinity for CBP/p300; therefore, they should be useful in studies investigating the biological roles of CBP and p300 and to validate the CBP and p300 bromodomains as therapeutic targets.
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Synaptophysin and synaptojanin-1 in Down syndrome are differentially affected by Alzheimer's disease.
J. Alzheimers Dis.
PUBLISHED: 06-15-2014
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Adults with Down syndrome (DS) develop Alzheimer's disease (AD) neuropathology by 40 years of age. Synaptophysin (SYN) consistently declines with age and is further reduced with sporadic AD. Thus, we hypothesized that SYN would be reduced in DS with AD. The gene for synaptojanin-1 (SYNJ1), involved in synaptic vesicle recycling, is on chromosome 21. We measured SYN and SYNJ1 in an autopsy series of 39 cases with DS and 28 without DS, along with 7 sporadic AD cases. SYN was significantly lower in DSAD compared with DS alone and similar to sporadic AD. Reduced SYN is associated with AD neuropathology and with A? levels in DS, as is seen in sporadic AD. SYNJ1 was significantly higher in DS and correlated with several measures of A?. SYNJ1 was higher in DSAD and significantly higher than SYNJ1 in sporadic AD. Although significantly higher in DS, SYNJ1 is further increased with AD neuropathology suggesting interesting differences in a synapse-associated protein that is overexpressed in trisomy 21.
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Notes From the Field: The Evaluation of Maine Nutrition and Physical Activity Self-Assessment for Child Care (NAPSACC) Experience.
Eval Health Prof
PUBLISHED: 05-30-2014
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More than half of all children in the United States aged 3 to 6 years are enrolled in child care centers. Maine received funds from the U.S. Department of Health and Human Services' Communities Putting Prevention to Work to promote the adoption of Nutrition and Physical Activity Self-Assessment for Child Care (NAPSACC), an evidence-based program for the child care setting. We evaluated the rollout and adoption of NAP SACC in Maine using multiple methods. Our findings suggest that the NAP SACC program has been successfully adopted in Maine. Nutrition and physical activity policies and offerings have improved, especially with regard to purchasing healthier options in the child care setting.
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Exploiting DNA mismatch repair deficiency as a therapeutic strategy.
Exp. Cell Res.
PUBLISHED: 05-28-2014
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The DNA Mismatch repair (MMR) pathway is critical for the maintenance of genomic stability. It is primarily responsible for the recognition and repair of mismatches that occur during DNA replication, but accumulating evidence suggest additional non-canonical roles for MMR proteins. MMR deficiency is a common feature of many tumor types. Germline mutations in MMR genes gives rise to the familial disorder, Lynch syndrome, which is associated with an increased predisposition to numerous cancers, including colorectal and endometrial. MMR deficiency has been associated with resistance to a wide range of standard therapeutic agents such as methylating agents, platinum compounds and fluoropyrimidine agents. Therefore, there is critical clinical need to identify new therapies for these resistant tumors. Recent studies, focussing on synthetic lethal interactions with MMR loss and emerging data identifying novel regulators of MMR may enable more successful treatment for MMR deficient patients. This review focuses on MMR loss in cancer and how exploiting both the canonical and non-canonical roles of MMR proteins may aid future therapeutic strategies.
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DNA mismatch repair and oxidative DNA damage: implications for cancer biology and treatment.
Cancers (Basel)
PUBLISHED: 05-14-2014
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Many components of the cell, including lipids, proteins and both nuclear and mitochondrial DNA, are vulnerable to deleterious modifications caused by reactive oxygen species. If not repaired, oxidative DNA damage can lead to disease-causing mutations, such as in cancer. Base excision repair and nucleotide excision repair are the two DNA repair pathways believed to orchestrate the removal of oxidative lesions. However, recent findings suggest that the mismatch repair pathway may also be important for the response to oxidative DNA damage. This is particularly relevant in cancer where mismatch repair genes are frequently mutated or epigenetically silenced. In this review we explore how the regulation of oxidative DNA damage by mismatch repair proteins may impact on carcinogenesis. We discuss recent studies that identify potential new treatments for mismatch repair deficient tumours, which exploit this non-canonical role of mismatch repair using synthetic lethal targeting.
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The proteomic response in glioblastoma in young patients.
J. Neurooncol.
PUBLISHED: 05-04-2014
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Increasing age is an important prognostic variable in glioblastoma (GBM). We have defined the proteomic response in GBM samples from 7 young patients (mean age 36 years) compared to peritumoural-control samples from 10 young patients (mean age 32 years). 2-Dimensional-gel-electrophoresis, image analysis, and protein identification (LC/MS) were performed. 68 proteins were significantly altered in young GBM samples with 29 proteins upregulated and 39 proteins downregulated. Over 50 proteins are described as altered in GBM for the first time. In a parallel analysis in old GBM (mean age 67 years), an excellent correlation could be demonstrated between the proteomic profile in young GBM and that in old GBM patients (r(2) = 0.95) with only 5 proteins altered significantly (p < 0.01). The proteomic response in young GBM patients highlighted alterations in protein-protein interactions in the immunoproteosome, NFkB signalling, and mitochondrial function and the same systems participated in the responses in old GBM patients.
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Interactions among mitochondrial proteins altered in glioblastoma.
J. Neurooncol.
PUBLISHED: 03-29-2014
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Mitochondrial dysfunction is putatively central to glioblastoma (GBM) pathophysiology but there has been no systematic analysis in GBM of the proteins which are integral to mitochondrial function. Alterations in proteins in mitochondrial enriched fractions from patients with GBM were defined with label-free liquid chromatography mass spectrometry. 256 mitochondrially-associated proteins were identified in mitochondrial enriched fractions and 117 of these mitochondrial proteins were markedly (fold-change ? 2) and significantly altered in GBM (p ? 0.05). Proteins associated with oxidative damage (including catalase, superoxide dismutase 2, peroxiredoxin 1 and peroxiredoxin 4) were increased in GBM. Protein-protein interaction analysis highlighted a reduction in multiple proteins coupled to energy metabolism (in particular respiratory chain proteins, including 23 complex-I proteins). Qualitative ultrastructural analysis in GBM with electron microscopy showed a notably higher prevalence of mitochondria with cristolysis in GBM. This study highlights the complex mitochondrial proteomic adjustments which occur in GBM pathophysiology.
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A series of potent CREBBP bromodomain ligands reveals an induced-fit pocket stabilized by a cation-? interaction.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 02-24-2014
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The benzoxazinone and dihydroquinoxalinone fragments were employed as novel acetyl lysine mimics in the development of CREBBP bromodomain ligands. While the benzoxazinone series showed low affinity for the CREBBP bromodomain, expansion of the dihydroquinoxalinone series resulted in the first potent inhibitors of a bromodomain outside the BET family. Structural and computational studies reveal that an internal hydrogen bond stabilizes the protein-bound conformation of the dihydroquinoxalinone series. The side chain of this series binds in an induced-fit pocket forming a cation-? interaction with R1173 of CREBBP. The most potent compound inhibits binding of CREBBP to chromatin in U2OS cells.
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Measurement of lysophospholipid acyltransferase activities using substrate competition.
J. Lipid Res.
PUBLISHED: 02-21-2014
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Lysophospholipid acyltransferases (LPATs) incorporate fatty acyl chains into phospholipids via a CoA-dependent mechanism and are important in remodeling phospholipids to generate the molecular species of phospholipids found in cells. These enzymes use one lysophospholipid and one acyl-CoA ester as substrates. Traditional enzyme activity assays engage a single substrate pair, whereas in vivo multiple molecular species exist. We describe here an alternative biochemical assay that provides a mixture of substrates presented to the microsomal extracts. Microsomal preparations from RAW 264.7 cells were used to compare traditional LPAT assays with data obtained using a dual substrate choice assay using six different lysophospholipids and eight different acyl-CoA esters. The complex mixture of newly synthesized phospholipid products was analyzed using LC-MS/MS. Both types of assays provided similar results, but the dual choice assay provided information about multiple fatty acyl chain incorporation into various phospholipid classes in a single reaction. Engineered suppression of LPCAT3 activity in RAW 264.7 cells was easily detected by the dual choice method. These findings demonstrate that this assay is both specific and sensitive and that it provides much richer biochemical detail than traditional assays.
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Cofilin drives cell-invasive and metastatic responses to TGF-? in prostate cancer.
Cancer Res.
PUBLISHED: 02-07-2014
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Cofilin (CFL) is an F-actin-severing protein required for the cytoskeleton reorganization and filopodia formation, which drives cell migration. CFL binding and severing of F-actin is controlled by Ser3 phosphorylation, but the contributions of this step to cell migration during invasion and metastasis of cancer cells are unclear. In this study, we addressed the question in prostate cancer cells, including the response to TGF-?, a critical regulator of migration. In cells expressing wild-type CFL, TGF-? treatment increased LIMK-2 activity and cofilin phosphorylation, decreasing filopodia formation. Conversely, constitutively active CFL (SerAla) promoted filipodia formation and cell migration mediated by TGF-?. Notably, in cocultures of prostate cancer epithelial cells and cancer-associated fibroblasts, active CFL promoted invasive migration in response to TGF-? in the microenvironment. Further, constitutively active CFL elevated the metastatic ability of prostate cancer cells in vivo. We found that levels of active CFL correlated with metastasis in a mouse model of prostate tumor and that in human prostate cancer, CFL expression was increased significantly in metastatic tumors. Our findings show that the actin-severing protein CFL coordinates responses to TGF-? that are needed for invasive cancer migration and metastasis.
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Luminal subtypes predict improved survival following central nervous system metastasis in patients with surgically managed metastatic breast carcinoma.
Arch. Pathol. Lab. Med.
PUBLISHED: 01-31-2014
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Metastatic breast cancer to the central nervous system (CNS) is second only to lung cancer metastasis to the CNS in frequency. Patients with triple-negative primary breast cancer and those with human epidermal growth factor receptor 2 (HER2)-positive primary breast cancer are at an increased risk for metastasis. Very little is known about predictive or prognostic variables once patients develop CNS metastases. Currently, therapeutic options are limited, with surgery generally offered primarily to those with solitary lesions.
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The reduced kinome of Ostreococcus tauri: core eukaryotic signalling components in a tractable model species.
BMC Genomics
PUBLISHED: 01-24-2014
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The current knowledge of eukaryote signalling originates from phenotypically diverse organisms. There is a pressing need to identify conserved signalling components among eukaryotes, which will lead to the transfer of knowledge across kingdoms. Two useful properties of a eukaryote model for signalling are (1) reduced signalling complexity, and (2) conservation of signalling components. The alga Ostreococcus tauri is described as the smallest free-living eukaryote. With less than 8,000 genes, it represents a highly constrained genomic palette.
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Dual kinase-bromodomain inhibitors for rationally designed polypharmacology.
Nat. Chem. Biol.
PUBLISHED: 01-23-2014
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Concomitant inhibition of multiple cancer-driving kinases is an established strategy to improve the durability of clinical responses to targeted therapies. The difficulty of discovering kinase inhibitors with an appropriate multitarget profile has, however, necessitated the application of combination therapies, which can pose major clinical development challenges. Epigenetic reader domains of the bromodomain family have recently emerged as new targets for cancer therapy. Here we report that several clinical kinase inhibitors also inhibit bromodomains with therapeutically relevant potencies and are best classified as dual kinase-bromodomain inhibitors. Nanomolar activity on BRD4 by BI-2536 and TG-101348, which are clinical PLK1 and JAK2-FLT3 kinase inhibitors, respectively, is particularly noteworthy as these combinations of activities on independent oncogenic pathways exemplify a new strategy for rational single-agent polypharmacological targeting. Furthermore, structure-activity relationships and co-crystal structures identify design features that enable a general platform for the rational design of dual kinase-bromodomain inhibitors.
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The pathologic spectrum of oculoleptomeningeal amyloidosis with Val30Gly transthyretin gene mutation in a postmortem case.
Hum. Pathol.
PUBLISHED: 01-02-2014
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We report the clinical and postmortem pathologic features of a 60-year-old woman with oculoleptomeningeal amyloidosis with a Val30Gly transthyretin gene mutation. Unlike other forms of hereditary amyloidosis, this rare type displays amyloid deposition predominantly in the eyes and central nervous system. Our patient belongs to 1 of only 2 kindreds known to carry this transthyretin mutation. Previous reports focused on examination of the brain and spinal cord, largely ignoring postmortem examination of the eyes. In this case, autopsy examination revealed amyloid deposition in the leptomeninges surrounding the brain, spinal cord, and optic nerves. Subependymal amyloid deposits projecting into the lateral ventricles as well as amyloid deposition in the choroid plexus, retinal vessels, nerve fiber layer of the retina, and vitreous were observed. Amyloid was not identified elsewhere in the body. Awareness of this rare form of hereditary amyloidosis is crucial, given the substantial genetic and therapeutic implications of the diagnosis. Oculoleptomeningeal amyloidosis can be easily diagnosed during life with vitreous biopsy, as was the case in our patient.
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Cytoskeleton targeting value in prostate cancer treatment.
Am J Clin Exp Urol
PUBLISHED: 01-01-2014
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Prostate cancer is a disease that affects hundreds of thousands of men in the United States each year. In the early stages of advanced prostate cancer, the disease can be suppressed by androgen deprivation therapy (ADT). Eventually, however, most patients experience resistance to androgen deprivation, and their treatment transitions to alternative targeting of the androgen axis with abiraterone and enzalutamide, as well as taxane-based chemotherapy. Development of advanced castration-resistant prostate cancer (CRPC) is a consequence of lack of an apoptotic response by the tumor cells to treatment. Understanding the mechanisms contributing to prostate tumor therapeutic resistance and progression to metastasis requires dissection of the signaling mechanisms navigating tumor invasion and metastasis as mediated by cell-matrix interactions engaging components of the extracellular matrix (ECM), to form adhesion complexes. For a tumor call to metastasize from the primary tumor, it requires disruption of cell-cell interactions from the surrounding cells, as well as detachment from the ECM and resistance to anoikis (apoptosis upon cell detachment from ECM). Attachment, movement and invasion of cancer cells are functionally facilitated by the actin cytoskeleton and tubulin as the structural component of microtubules. Transforming growth factor (TGF)-? has tumor-inhibitory activity in the early stages of tumorigenesis, but it promotes tumor invasive characteristics in metastatic disease. Recent evidence implicates active (dephosphorylated) cofilin, an F-actin severing protein required for cytoskeleton reorganization, as an important contributor to switching TGF-? characteristics from a growth suppressor to a promoter of prostate cancer invasion and metastasis. Cancer cells eventually lose the ability to adhere to adjacent neighboring cells as well as ECM proteins, and via epithelial-mesenchymal transition (EMT), acquire invasive and metastatic characteristics. Microtubule-targeting chemotherapeutic agents, taxanes, are used in combination with antiandrogen strategies to increase the survival rate in patients with CRPC. This review addresses the development of therapeutic platform for targeting the integrity of actin cytoskeleton to impair prostate cancer progression.
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Maternal high-fat diet worsens memory deficits in the triple-transgenic (3xTgAD) mouse model of Alzheimer's disease.
PLoS ONE
PUBLISHED: 01-01-2014
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Alzheimer's disease (AD) is not normally diagnosed until later in life, although evidence suggests that the disease starts at a much earlier age. Risk factors for AD, such as diabetes, hypertension and obesity, are known to have their affects during mid-life, though events very early in life, including maternal over-nutrition, can predispose offspring to develop these conditions. This study tested whether over-nutrition during pregnancy and lactation affected the development of AD in offspring, using a transgenic AD mouse model. Female triple-transgenic AD dam mice (3xTgAD) were exposed to a high-fat (60% energy from fat) or control diet during pregnancy and lactation. After weaning (at 3 weeks of age), female offspring were placed on a control diet and monitored up until 12 months of age during which time behavioural tests were performed. A transient increase in body weight was observed in 4-week-old offspring 3xTgAD mice from dams fed a high-fat diet. However, by 5 weeks of age the body weight of 3xTgAD mice from the maternal high-fat fed group was no different when compared to control-fed mice. A maternal high-fat diet led to a significant impairment in memory in 2- and 12-month-old 3xTgAD offspring mice when compared to offspring from control fed dams. These effects of a maternal high-fat diet on memory were accompanied by a significant increase (50%) in the number of tau positive neurones in the hippocampus. These data demonstrate that a high-fat diet during pregnancy and lactation increases memory impairments in female 3xTgAD mice and suggest that early life events during development might influence the onset and progression of AD later in life.
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[1,2,4]Triazolo[4,3-a]phthalazines: Inhibitors of Diverse Bromodomains.
J. Med. Chem.
PUBLISHED: 12-30-2013
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Bromodomains are gaining increasing interest as drug targets. Commercially sourced and de novo synthesized substituted [1,2,4]triazolo[4,3-a]phthalazines are potent inhibitors of both the BET bromodomains such as BRD4 as well as bromodomains outside the BET family such as BRD9, CECR2, and CREBBP. This new series of compounds is the first example of submicromolar inhibitors of bromodomains outside the BET subfamily. Representative compounds are active in cells exhibiting potent cellular inhibition activity in a FRAP model of CREBBP and chromatin association. The compounds described are valuable starting points for discovery of selective bromodomain inhibitors and inhibitors with mixed bromodomain pharmacology.
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Designer Psychostimulants in Urine by Liquid Chromatography-Tandem Mass Spectrometry.
J. Forensic Sci.
PUBLISHED: 12-06-2013
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Designer psychostimulants are known by recreational drug users to produce a complex array of adrenergic and hallucinogenic effects. Many of these drugs are not targeted during routine toxicology testing and as a consequence, they are rarely reported. The purpose of this study was to develop a procedure for the detection of 15 psychostimulants in urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS), specifically 2,5-dimethoxy-4-bromophenethylamine (2C-B), 2,5-dimethoxy-4-chlorophenethylamine (2C-C), 2,5-dimethoxy-4-methylphenethylamine (2C-D), 2,5-dimethoxy-4-ethylphenethylamine (2C-E), 2,5-dimethoxyphenethylamine (2C-H), 2,5-dimethoxy-4-iodophenethylamine (2C-I), 2,5-dimethoxy-4-ethylthiophenethylamine (2C-T-2), 2,5-dimethoxy-4-isopropylthiophenethylamine (2C-T-4), 2,5-dimethoxy-4-propylthiophenethylamine (2C-T-7), 2,5-dimethoxy-4-bromoamphetamine (DOB), 2,5-dimethoxy-4-chloroamphetamine (DOC), 2,5-dimethoxy-4-ethylamphetamine (DOET), 2,5-dimethoxy-4-iodoamphetamine (DOI), 2,5-dimethoxy-4-methylamphetamine (DOM), and 4-methylthioamphetamine (4-MTA). Analytical recoveries using solid-phase extraction were 64-92% and the limit of detection was 0.5 ng/mL for all drugs except 2C-B (1 ng/mL). The assay was evaluated in terms of analytical recovery, precision, accuracy, linearity, matrix effect, and interferences. The technique allows for the simultaneous detection of 15 psychostimulants at sub-ng/mL concentrations.
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RVX-208, an inhibitor of BET transcriptional regulators with selectivity for the second bromodomain.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 11-18-2013
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Bromodomains have emerged as attractive candidates for the development of inhibitors targeting gene transcription. Inhibitors of the bromo and extraterminal (BET) family recently showed promising activity in diverse disease models. However, the pleiotropic nature of BET proteins regulating tissue-specific transcription has raised safety concerns and suggested that attempts should be made for domain-specific targeting. Here, we report that RVX-208, a compound currently in phase II clinical trials, is a BET bromodomain inhibitor specific for second bromodomains (BD2s). Cocrystal structures revealed binding modes of RVX-208 and its synthetic precursor, and fluorescent recovery after photobleaching demonstrated that RVX-208 displaces BET proteins from chromatin. However, gene-expression data showed that BD2 inhibition only modestly affects BET-dependent gene transcription. Our data demonstrate the feasibility of specific targeting within the BET family resulting in different transcriptional outcomes and highlight the importance of BD1 in transcriptional regulation.
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Extravasation management of nonchemotherapeutic medications.
J Infus Nurs
PUBLISHED: 11-09-2013
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Many who think of vesicants think of chemotherapy and oncology patients. But not all vesicants are chemotherapy medications. This article reviews the factors that increase the risk of extravasation, the nonchemotherapeutic medications associated with extravasation injuries, and the recommended treatments.
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Giant Cell Arteritis Presenting with Uveitis.
Ocul. Immunol. Inflamm.
PUBLISHED: 10-21-2013
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Abstract Giant cell arteritis, also known as temporal arteritis, is the most common primary vasculitis affecting the nervous system. Early recognition of this treatable condition is essential to avoid potentially devastating complications. Giant cell arteritis occurs in adults older than 50 years and affects large and medium-sized arteries, especially the external and internal carotid arteries and their branches. Severe inflammation of the vessel wall may result in obstruction of the lumen and end-organ ischemia. Typical giant cell arteritis symptoms include headache, scalp tenderness, jaw claudication, and polymyalgia rheumatica. Ischemia induced by the arteritis can lead to blindness. Herein, we describe a rare case of giant cell arteritis in a patient who initially presented with uveitis, thus eluding timely diagnosis and prompt therapy.
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Discovery of Novel Small-Molecule Inhibitors of BRD4 Using Structure-Based Virtual Screening.
J. Med. Chem.
PUBLISHED: 10-03-2013
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Bromodomains (BRDs) are epigenetic readers that recognize acetylated-lysine (KAc) on proteins and are implicated in a number of diseases. We describe a virtual screening approach to identify BRD inhibitors. Key elements of this approach are the extensive design and use of substructure queries to compile a set of commercially available compounds featuring novel putative KAc mimetics and docking this set for final compound selection. We describe the validation of this approach by applying it to the first BRD of BRD4. The selection and testing of 143 compounds lead to the discovery of six novel hits, including four unprecedented KAc mimetics. We solved the crystal structure of four hits, determined their binding mode, and improved their potency through synthesis and the purchase of derivatives. This work provides a validated virtual screening approach that is applicable to other BRDs and describes novel KAc mimetics that can be further explored to design more potent inhibitors.
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Sequential analysis of mothers and fathers reassurance and childrens postoperative distress.
J Pediatr Psychol
PUBLISHED: 08-20-2013
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Children experience distress after surgery. Associations exist between parent reassurance (e.g., "Its OK") and child distress, but little is known about the causal direction of these interactions. This study examined sequential relations between mothers and fathers reassurance and childrens distress.
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Functional analysis of the rodent CK1tau mutation in the circadian clock of a marine unicellular alga.
BMC Cell Biol.
PUBLISHED: 07-29-2013
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Casein Kinase 1 (CK1) is one of few proteins known to affect cellular timekeeping across metazoans, and the naturally occurring CK1tau mutation shortens circadian period in mammals. Functional conservation of a timekeeping function for CK1 in the green lineage was recently identified in the green marine unicell Ostreococcus tauri, in spite of the absence of CK1s transcriptional targets known from other species. The short-period phenotype of CK1tau mutant in mammals depends specifically on increased CK1 activity against PERIOD proteins. To understand how CK1 acts differently upon the algal clock, we analysed the cellular and proteomic effects of CK1tau overexpression in O. tauri.
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Arsenic is cytotoxic and genotoxic to primary human lung cells.
Mutat. Res.
PUBLISHED: 05-02-2013
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Arsenic originates from both geochemical and numerous anthropogenic activities. Exposure of the general public to significant levels of arsenic is widespread. Arsenic is a well-documented human carcinogen. Long-term exposure to high levels of arsenic in drinking water has been linked to bladder, lung, kidney, liver, prostate, and skin cancers. Among them, lung cancer is of great public concern. However, little is known about how arsenic causes lung cancer and few studies have considered effects in normal human lung cells. The purpose of this study was to determine the cytotoxicity and genotoxicity of arsenic in human primary bronchial fibroblast and epithelial cells. Our data show that arsenic induces a concentration-dependent decrease in cell survival after short (24h) or long (120h) exposures. Arsenic induces concentration-dependent but not time-dependent increases in chromosome damage in fibroblasts. No chromosome damage is induced after either 24h or 120h arsenic exposure in epithelial cells. Using neutral comet assay and gamma-H2A.X foci forming assay, we found that 24h or 120h exposure to arsenic induces increases in DNA double strand breaks in both cell lines. These data indicate that arsenic is cytotoxic and genotoxic to human lung primary cells but lung fibroblasts are more sensitive to arsenic than epithelial cells. Further research is needed to understand the specific mechanisms involved in arsenic-induced genotoxicity in human lung cells.
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Optimization of 3,5-dimethylisoxazole derivatives as potent bromodomain ligands.
J. Med. Chem.
PUBLISHED: 04-05-2013
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The bromodomain protein module, which binds to acetylated lysine, is emerging as an important epigenetic therapeutic target. We report the structure-guided optimization of 3,5-dimethylisoxazole derivatives to develop potent inhibitors of the BET (bromodomain and extra terminal domain) bromodomain family with good ligand efficiency. X-ray crystal structures of the most potent compounds reveal key interactions required for high affinity at BRD4(1). Cellular studies demonstrate that the phenol and acetate derivatives of the lead compounds showed strong antiproliferative effects on MV4;11 acute myeloid leukemia cells, as shown for other BET bromodomain inhibitors and genetic BRD4 knockdown, whereas the reported compounds showed no general cytotoxicity in other cancer cell lines tested.
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Putting evidence into nursing practice: four traditional practices not supported by the evidence.
Crit Care Nurse
PUBLISHED: 04-03-2013
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Evidence-based nursing practice is essential to the delivery of high-quality care that optimizes patients outcomes. Studies continue to show improved outcomes when best evidence is used in the delivery of patient care. Despite awareness of the importance of practicing by using best evidence, achieving and sustaining evidence-based practice within practice environments can be challenging, and research suggests that integration of evidence-based practice into daily clinical practice remains inconsistent. This article addresses 4 practice issues that, first, are within the realm of nursing and if changed might improve care of patients and, second, are areas in which the tradition and the evidence do not agree and practice continues to follow tradition. The topics addressed are (1) noninvasive measurement of blood pressure in children, (2) oxygen administration for patients with chronic obstructive pulmonary disease, (3) intravenous catheter size and blood administration, and (4) infection control practices to prevent infections. The related beliefs, current evidence, and recommendations for practice related to each topic are described.
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Prenatal attitudes and parity predict selection into a U.S. child health program: a short report.
Soc Sci Med
PUBLISHED: 03-05-2013
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Public policies are a determinant of child health disparities; sound evaluation of these programs is essential for good governance. It is impossible in most countries to randomize assignment into child health programs that directly offer benefits. In the absence of this, researchers face the threat of selection bias-the idea that there are innate, immeasurable differences between those who take-up treatment and those who dont. In the field of Program Evaluation we are most concerned with the differences between the eligible people who take-up a program and the eligible people who choose not to enroll. Using a case study of a large U.S. nutrition program, this report illustrates how the perceived benefits of participation may affect the decision to take-up a program. In turn, this highlights sources of potential selection bias. Using data from a longitudinal study of mothers and infants conducted between May and December of 2005, I show that attitudes and beliefs prenatally toward breastfeeding determine enrollment in a U.S nutrition program that offers free Infant Formula. I also find that the significance of the selection bias differs by parity. Analysis reveals that maternal attitudinal responses are more highly predictive of future behavior, compared to standard demographic variables. In sum, this paper makes a case for rigorously understanding the factors that determine take-up of a program and how those factors can modify the results of a program evaluation.
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Marginal zone B-cell lymphoma involving a longstanding fibrous meningioma: an initial manifestation of systemic disease.
Hum. Pathol.
PUBLISHED: 02-22-2013
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The combined presence of meningioma and lymphoma involving the dura is exceptionally rare. A 62-year-old woman, radiologically diagnosed with meningioma 14 years prior but never treated, presented with headaches and visual symptoms. Magnetic resonance imaging demonstrated significant growth of the mass. Surgical resection yielded a composite meningioma and marginal zone B-cell lymphoma. Subsequent systemic workup revealed bone marrow involvement. Low-grade lymphomas rarely metastasize to the central nervous system. When they do, it is usually a result of large cell transformation and typically marks a late event in the course of the disease. This case highlights the necessity of adequate sampling of meningiomas and of including low-grade lymphoma in the differential diagnosis of meningiomas with prominent lymphocytic infiltrates. In addition, this case emphasizes that all patients with lymphoma involving the central nervous system, even when low grade, should receive a full systemic workup.
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MALDI MS imaging analysis of apolipoprotein E and lysyl oxidase-like 1 in human lens capsules affected by pseudoexfoliation syndrome.
J Proteomics
PUBLISHED: 01-09-2013
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Pseudoexfoliation (PEX) syndrome is an age-related systemic disease of the extracellular matrix, characterized by the presence of amyloid-like fibrillar deposits on the anterior lens capsule. The pathological deposits (PEX material) can obstruct aqueous outflow leading to increased intraocular pressure that in turn can result in glaucoma. PEX syndrome is the most common risk factor for glaucoma. In our previous work, we reported a protocol for the analysis of human lens capsules by MALDI MS imaging. Here, we extend our previous work applying the developed protocol to the analysis of human lens capsules affected by PEX syndrome. We focus our investigation on known components of the PEX material, namely lysyl oxidase-like 1 (LOXL1) and apolipoprotein E (APOE). Our results show that LOXL1 is more abundant in the deposits in the iris region and, alternatively APOE is concentrated in the PEX material accumulated in the pupillary area of the anterior lens capsule. Furthermore, we identify potentially relevant post-translational modifications which may have an important role in promoting the cross-linking processes in PEX syndrome and stabilize aggregate structures within the proteinaceous PEX material.
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Functional analysis of Casein Kinase 1 in a minimal circadian system.
PLoS ONE
PUBLISHED: 01-01-2013
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The Earths rotation has driven the evolution of cellular circadian clocks to facilitate anticipation of the solar cycle. Some evidence for timekeeping mechanism conserved from early unicellular life through to modern organisms was recently identified, but the components of this oscillator are currently unknown. Although very few clock components appear to be shared across higher species, Casein Kinase 1 (CK1) is known to affect timekeeping across metazoans and fungi, but has not previously been implicated in the circadian clock in the plant kingdom. We now show that modulation of CK1 function lengthens circadian rhythms in Ostreococcustauri, a unicellular marine algal species at the base of the green lineage, separated from humans by ~1.5 billion years of evolution. CK1 contributes to timekeeping in a phase-dependent manner, indicating clock-mediated gating of CK1 activity. Label-free proteomic analyses upon overexpression as well as inhibition revealed CK1-responsive phosphorylation events on a set of target proteins, including highly conserved potentially clock-relevant cellular regulator proteins. These results have major implications for our understanding of cellular timekeeping and can inform future studies in any circadian organism.
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Childrens behavior in the postanesthesia care unit: the development of the Child Behavior Coding System-PACU (CBCS-P).
J Pediatr Psychol
PUBLISHED: 12-13-2011
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To develop and validate a behavioral coding measure, the Childrens Behavior Coding System-PACU (CBCS-P), for childrens distress and nondistress behaviors while in the postanesthesia recovery unit.
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Glioblastoma occurring at the site of a previous medulloblastoma following a 5-year remission period.
Neuropathology
PUBLISHED: 12-08-2011
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We describe a case of a 14-year-old boy who developed a cerebellar and brainstem glioblastoma 5 years after treatment for a medulloblastoma. The patient first presented in 2003 with 9 months of vomiting and a 9-kg weight loss. A head MRI showed a heterogeneously enhancing posterior fossa mass with hydrocephalus. Gross total resection was performed and the tumor was consistent with a classic medulloblastoma. Postoperative chemotherapy and craniospinal radiation was administered. The patient remained tumor-free until 2008, at which time he presented with right-sided weakness and numbness, left eye pain, vomiting and weight loss. Imaging showed abnormalities within the posterior pons, medulla, inferior cerebellar peduncles, cerebellar hemispheres and cervicomedullary junction with expansion of the medulla and cervical spinal cord. Due to the location of the lesion, biopsy was felt to be too risky and was avoided. Despite receiving chemotherapy, his symptoms continued to worsen and he died 4 months later. Post mortem examination limited to the brain and spinal cord confirmed the radiographic extent of the tumor. Microscopic examination showed a highly cellular infiltrative glial neoplasm with extensive palisading necrosis. A diagnosis of glioblastoma was rendered. The question of whether the first and second tumors were related is of potential clinical and academic interest. The first tumor was synaptophysin-positive and GFAP-negative, consistent with medulloblastoma. The second tumor was synaptophysin-negative and focally GFAP-positive, consistent with glioblastoma. The glioblastoma displayed EGF receptor amplification, and interestingly, it also displayed MYCN amplification; both tumors showed low level PTEN deletion. The medulloblastoma displayed a signal pattern consistent with an isochromosome 17q, while the glioblastoma showed some cells with an isochromosome 17q signal pattern amid a background of cells with abundant chromosomal instability. The relationship between these two tumors, particularly with regard to various molecular events, is discussed.
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Proteome turnover in the green alga Ostreococcus tauri by time course 15N metabolic labeling mass spectrometry.
J. Proteome Res.
PUBLISHED: 12-01-2011
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Protein synthesis and degradation determine the cellular levels of proteins, and their control hence enables organisms to respond to environmental change. Experimentally, these are little known proteome parameters; however, recently, SILAC-based mass spectrometry studies have begun to quantify turnover in the proteomes of cell lines, yeast, and animals. Here, we present a proteome-scale method to quantify turnover and calculate synthesis and degradation rate constants of individual proteins in autotrophic organisms such as algae and plants. The workflow is based on the automated analysis of partial stable isotope incorporation with (15)N. We applied it in a study of the unicellular pico-alga Ostreococcus tauri and observed high relative turnover in chloroplast-encoded ATPases (0.42-0.58% h(-1)), core photosystem II proteins (0.34-0.51% h(-1)), and RbcL (0.47% h(-1)), while nuclear-encoded RbcS2 is more stable (0.23% h(-1)). Mitochondrial targeted ATPases (0.14-0.16% h(-1)), photosystem antennae (0.09-0.14% h(-1)), and histones (0.07-0.1% h(-1)) were comparatively stable. The calculation of degradation and synthesis rate constants k(deg) and k(syn) confirms RbcL as the bulk contributor to overall protein turnover. This study performed over 144 h of incorporation reveals dynamics of protein complex subunits as well as isoforms targeted to different organelles.
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Organ-distribution of the metabolite 2-aminothiazoline-4-carboxylic acid in a rat model following cyanide exposure.
Biomarkers
PUBLISHED: 10-24-2011
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The reaction of cyanide (CN(-)) with cystine to produce 2-aminothiazoline-4-carboxylic acid (ATCA) is one of the independent detoxification pathways of cyanide in biological systems. In this report, in vivo production of ATCA and its distributions in plasma and organs were studied after a subcutaneous sublethal dose of 4?mg/kg body weight potassium cyanide (KCN) administration to rats. At this sublethal dose of KCN, ATCA concentration was not significantly increased in the plasma samples, however, it was found significantly increased in liver samples. These results suggested that ATCA might not be a good diagnostic biomarker in plasma for sublethal cyanide exposure; however, liver could serve as the right organ for the detection of ATCA in post-mortem examinations involving cyanide exposure in military, firefighting, industrial and forensic settings.
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Clinician perspectives on barriers to and opportunities for skin-to-skin contact for premature infants in neonatal intensive care units.
Breastfeed Med
PUBLISHED: 10-19-2011
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Our objective was to investigate key factors in promoting skin-to-skin contact (STSC) in the neonatal intensive care unit (NICU).
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Proteomic analysis of mitochondria in APOE transgenic mice and in response to an ischemic challenge.
J. Cereb. Blood Flow Metab.
PUBLISHED: 08-31-2011
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Apolipoprotein E (APOE)-?4 is associated with a deleterious outcome after ischemic brain injury, which may involve abnormal regulation of mitochondrial function. We have assessed the mitochondrial proteomic response of APOE-?3 and APOE-?4 transgenic mice to transient global ischemic injury in the hippocampus. A genotype-dependent increase in ApoE levels in mitochondria was observed after ischemia, with APOE-?4 mice showing significantly greater increases than APOE-?3 mice. Quantitative analysis of the mitochondria-enriched fractions was performed using liquid-chromatography mass spectrometry coupled to label-free analysis. Of the 1,067 identified proteins, 274 were mitochondria associated. Mitochondrial protein expression was significantly different between genotypes under basal conditions as well as in response to global ischemia. A total of 12 mitochondrial proteins (including respiratory chain proteins NDUFA11, NDUFS3, NDUF5B, ATP5J, as well as ETFA, CYB5B, ATP6V1A, HSPA1B, OXR1, GLUL, IARS2, and PHYHIPL) were significantly altered with respect to genotype, global ischemia, or their interaction (P<0.01). A compelling interactome, created using proteins found to be significantly modulated by global ischemia (P<0.05), involved proteins that regulate energy production and oxidative stress. Thus, APOE genotype has a differential effect on the mitochondrial protein expression in the absence and presence of an injury, which may underlie the differing genotype susceptibility.
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Diglycolic acid is the nephrotoxic metabolite in diethylene glycol poisoning inducing necrosis in human proximal tubule cells in vitro.
Toxicol. Sci.
PUBLISHED: 08-18-2011
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Diethylene glycol (DEG), a solvent and chemical intermediate, can produce an acute toxic syndrome, the hallmark of which is acute renal failure due to cortical tubular degeneration and proximal tubular necrosis. DEG is metabolized to two primary metabolites, 2-hydroxyethoxyacetic acid (2-HEAA) and diglycolic acid (DGA), which are believed to be the proximate toxicants. The precise mechanism of toxicity has yet to be elucidated, so these studies were designed to determine which metabolite was responsible for the proximal tubule cell death. Human proximal tubule (HPT) cells in culture, obtained from normal cortical tissue and passaged 3-6 times, were incubated with increasing concentrations of DEG, 2-HEAA, or DGA separately and in combination for 48 h at pH 6 or 7.4, and various parameters of necrotic and apoptotic cell death were measured. DEG and 2-HEAA did not produce any cell death. DGA produced dose-dependent necrosis at concentrations above 25 mmol/l. DGA did not affect caspase-3 activity and increased annexin V staining only in propidium iodide-stained cells. Hence, DGA induced necrosis, not apoptosis, as corroborated by severe depletion of cellular adenosine triphosphate levels. DGA is structurally similar to citric acid cycle intermediates that are taken up by specific transporters in kidney cells. HPT cells, incubated with N-(p-amylcinnamoyl)anthranilic acid, a sodium dicarboxylate-1 transporter inhibitor showed significantly decreased cell death compared with DGA alone. These studies demonstrate that DGA is the toxic metabolite responsible for DEG-induced proximal tubular necrosis and suggest a possible transporter-mediated uptake of DGA leading to toxic accumulation and cellular dysfunction.
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Optimization of liposomal lipid composition for a new, reactive sulfur donor, and in vivo efficacy studies on mice to antagonize cyanide intoxication.
J Drug Deliv
PUBLISHED: 08-08-2011
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Present studies have focused on a novel cyanide antidotal system, on the coencapsulation of a new sulfur donor DTO with rhodanese within sterically stabilized liposomes. The optimal lipid composition for coencapsulation of DTO with rhodanese is the combination of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, cholesterol, cationic lipid (DOTAP), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] ammonium salt (with molar ratios of 82.7 : 9.2 : 3.0 : 5.1). With the optimized compositions, prophylactic and therapeutic in vivo efficacy studies were carried out in a mice model. When DTO was coencapsulated with rhodanese and thiosulfate the prophylactic antidotal protection was 4.9 × LD(50). Maximum antidotal protection against cyanide intoxication (15 × LD(50)) was achieved with coencapsulated rhodanese and DTO/thiosulfate in combination with sodium nitrite. When applied therapeutically, 100% survival rate (6/6) was achieved at 20?mg/kg cyanide doses with the encapsulated DTO-rhodanese-thiosulfate antidotal systems with and without sodium nitrite. These data are indicating that the appropriately formulated DTO is a promising sulfur donor for cyanide antagonism.
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MALDI-MS-imaging of whole human lens capsule.
J. Proteome Res.
PUBLISHED: 06-29-2011
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The ocular lens capsule is a smooth, transparent basement membrane that encapsulates the lens and is composed of a rigid network of interacting structural proteins and glycosaminoglycans. During cataract surgery, the anterior lens capsule is routinely removed in the form of a circular disk. We considered that the excised capsule could be easily prepared for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-MSI) analysis. MALDI-MSI is a powerful tool to elucidate the spatial distribution of small molecules, peptides, and proteins within tissues. Here, we apply this molecular imaging technique to analyze the freshly excised human lens capsule en face. We demonstrate that novel information about the distribution of proteins by MALDI-MSI can be obtained from this highly compact connective tissue, having no evident histo-morphological characteristics. Trypsin digestion carried out on-tissue is shown to improve MALDI-MSI analysis of human lens capsules and affords high repeatability. Most importantly, MALDI-MSI analysis reveals a concentric distribution pattern of proteins such as apolipoprotein E (ApoE) and collagen IV alpha-1 on the anterior surface of surgically removed lens capsule, which may indicate direct or indirect effects of environmental and mechanical stresses on the human ocular lens.
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Elevated stearoyl-CoA desaturase in brains of patients with Alzheimers disease.
PLoS ONE
PUBLISHED: 06-21-2011
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The molecular bases of Alzheimers disease (AD) remain unclear. We used a lipidomic approach to identify lipid abnormalities in the brains of subjects with AD (N = 37) compared to age-matched controls (N = 17). The analyses revealed statistically detectable elevations in levels of non-esterified monounsaturated fatty acids (MUFAs) and mead acid (20:3n-9) in mid-frontal cortex, temporal cortex and hippocampus of AD patients. Further studies showed that brain mRNAs encoding for isoforms of the rate-limiting enzyme in MUFAs biosynthesis, stearoyl-CoA desaturase (SCD-1, SCD-5a and SCD-5b), were elevated in subjects with AD. The monounsaturated/saturated fatty acid ratio (desaturation index)--displayed a strong negative correlation with measures of cognition: the Mini Mental State Examination test (r = -0.80; P = 0.0001) and the Boston Naming test (r = -0.57; P = 0.0071). Our results reveal a previously unrecognized role for the lipogenic enzyme SCD in AD.
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Cytoplasmic p63 immunohistochemistry is a useful marker for muscle differentiation: an immunohistochemical and immunoelectron microscopic study.
Mod. Pathol.
PUBLISHED: 05-27-2011
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TP63, a member of the TP53 gene family, is a nuclear marker of myoepithelial cells. Antibody against p63 is frequently used to aid in the diagnosis of prostate carcinoma, as well as in the identification of myoepithelial cells in other tissues including the breast. p63 is also a marker for squamous cell carcinoma. Recently, it was found that all p53 family members are involved in regulating the process of muscle differentiation through the retinoblastoma (RB) protein. Ablation of these p53 family functions blocks the differentiation program and promotes malignant transformation by enabling cooperating oncogenes to transform myoblasts. We therefore studied p63 expression in a number of neoplasms with myogenic differentiation. Immunohistochemical staining for p63 was performed on paraffin sections from 38 rhabdomyosarcomas, five leiomyomas, five leiomyosarcomas, five rhabdomyomas, five rhabdomyomatous Wilms tumors, three normal cardiac muscles, one medullomyoblastoma, one pleuropulmonary blastoma with rhabdomyomatous differentiation, and one teratoma with prominent rhabdomyoblasts. Each case was also stained with desmin. Unlike the nuclear staining scored in myoepithelial cells, only cytoplasmic staining for p63 was considered positive. Of 38 cases of rhabdomyosarcoma, 36 showed cytoplasmic p63 staining; 24 of these showed highlighting of cross-striations superior to that of desmin. In addition, 5/5 rhabdomyomas, 5/5 rhabdomyomatous Wilms tumors, 1/1 pleuropulmonary blastoma with rhabdomyomatous differentiation, 1/1 teratoma with atypical rhabdoblasts, and 1/1 medullomyoblastoma exhibited cytoplasmic p63 staining. Normal cardiac muscle samples (3/3) also demonstrated positive cytoplasmic staining and distinct cross-striations. Smooth muscle tumors exhibited only very focal and faint cytoplasmic staining in 5/5 leiomyomas and 4/5 leiomyosarcomas. Immunoelectron microscopic study of skeletal muscle showed p63 localization to the Z bands of sarcomeres. We conclude that p63 immunostain is a sensitive marker for skeletal muscle differentiation and highlights the cross-striations of strap cells with exceptional definition.
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Changing healthcare providers behavior during pediatric inductions with an empirically based intervention.
Anesthesiology
PUBLISHED: 05-25-2011
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Each year more than 4 million children experience significant levels of preoperative anxiety, which has been linked to poor recovery outcomes. Healthcare providers (HCPs) and parents represent key resources for children to help them manage their preoperative anxiety. The current study reports on the development and preliminary feasibility testing of a new intervention designed to change HCP and parent perioperative behaviors that have been reported previously to be associated with childrens coping and stress behaviors before surgery.
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Maximising student preparation for clinical teaching placements.
Clin Teach
PUBLISHED: 05-19-2011
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With the increasing emphasis on ambulatory health care, clinical educators need to ensure that students are sufficiently prepared to maximise learning opportunities during placements in ambulatory settings.
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Desire for perioperative information and parental ethnicity.
Paediatr Anaesth
PUBLISHED: 05-09-2011
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To examine the role of ethnicity, language, and socioeconomic variables in parental desire for information regarding childrens surgery. Aim:? To compare anesthetic and surgical information desired between English- and Spanish-speaking White and Hispanic mothers of children undergoing outpatient surgery.
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Preoperative anxiety in adolescents undergoing surgery: a pilot study.
Paediatr Anaesth
PUBLISHED: 04-25-2011
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The purpose of this study was to conduct a prospective assessment of preoperative anxiety in adolescents undergoing surgery.
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Parental attitudes regarding analgesic use for children: differences in ethnicity and language.
J. Pediatr. Surg.
PUBLISHED: 03-22-2011
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The aim of this study was to identify the impact of ethnicity and language on parental attitudes regarding analgesic use to treat childrens pain.
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Coenzyme Q10 and cognition in atorvastatin treated dogs.
Neurosci. Lett.
PUBLISHED: 03-21-2011
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Statins have been suggested to protect against Alzheimers disease (AD). Recently, however, we reported that aged dogs that underwent chronic statin treatment exhibited cognitive deficits compared with age matched controls. In human studies, blood levels of Coenzyme Q10 (CoQ10) decrease with statin use. CoQ10 is important for proper mitochondrial function and is a powerful antioxidant, two important factors for cognitive health in aging. Thus, the current study tested the hypothesis that CoQ10 levels in the serum and/or parietal cortex are decreased in statin treated dogs and are associated with poorer cognition. Six aged beagles (>8 years) were administered 80 mg/day of atorvastatin for 14.5 months and compared with placebo-treated animals. As predicted, serum CoQ10 was significantly lower in statin-treated dogs. Parietal cortex CoQ10 was not different between the two groups. However, poorer cognition was correlated with lower parietal cortex CoQ10. This study in dogs suggests that serum CoQ10 is reduced with atorvastatin treatment. CoQ10 levels in brain may be linked to impaired cognition in response to atorvastatin, in agreement with previous reports that statins may have a negative impact on cognition in the elderly.
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Shotgun proteomic analysis of the unicellular alga Ostreococcus tauri.
J Proteomics
PUBLISHED: 03-10-2011
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Ostreococcus tauri is a unicellular green alga and amongst the smallest and simplest free-living eukaryotes. The O. tauri genome sequence was determined in 2006. Molecular, physiological and taxonomic data that has been generated since then highlight its potential as a simple model species for algae and plants. However, its proteome remains largely unexplored. This paper describes the global proteomic study of O. tauri, using mass spectrometry-based approaches: phosphopeptide enrichment, cellular fractionation, label-free quantification and (15)N metabolic labeling. The O. tauri proteome was analyzed under the following conditions: sampling at different times during the circadian cycle, after 24h of illumination, after 24h of darkness and under various nitrogen source supply levels. Cell cycle related proteins such as dynamin and kinesin were significantly up-regulated during the daylight-to-darkness transition. This is reflected by their higher intensity at ZT13 and this transition phase coincides with the end of mitosis. Proteins involved in several metabolic mechanisms were found to be up-regulated under low nitrogen conditions, including carbon storage pathways, glycolysis, phosphate transport, and the synthesis of inorganic polyphosphates. Ostreococcus tauri responds to low nitrogen conditions by reducing its nitrogen assimilation machinery which suggests an atypical adaptation mechanism for coping with a nutrient-limited environment.
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Parallel high-throughput RNA interference screens identify PINK1 as a potential therapeutic target for the treatment of DNA mismatch repair-deficient cancers.
Cancer Res.
PUBLISHED: 01-17-2011
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Synthetic lethal approaches to cancer treatment have the potential to deliver relatively large therapeutic windows and therefore significant patient benefit. To identify potential therapeutic approaches for cancers deficient in DNA mismatch repair (MMR), we have carried out parallel high-throughput RNA interference screens using tumor cell models of MSH2- and MLH1-related MMR deficiency. We show that silencing of the PTEN-induced putative kinase 1 (PINK1), is synthetically lethal with MMR deficiency in cells with MSH2, MLH1, or MSH6 dysfunction. Inhibition of PINK1 in an MMR-deficient background results in an elevation of reactive oxygen species and the accumulation of both nuclear and mitochondrial oxidative DNA lesions, which likely limit cell viability. Therefore, PINK1 represents a potential therapeutic target for the treatment of cancers characterized by MMR deficiency caused by a range of different gene deficiencies.
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Therapeutic interventions targeting Beta amyloid pathogenesis in an aging dog model.
Curr Neuropharmacol
PUBLISHED: 01-13-2011
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Aged dogs and humans share complex cognitive and pathological responses to aging. Specifically, dogs develop Alzheimers Disease (AD) like beta-amyloid (A?) that are associated with cognitive deficits. Currently, therapeutic approaches to prevent AD are targeted towards reduced production, aggregation and increased clearance of A?. The current review discusses cognition and neuropathology of the aging canine model and how it has and continues to be useful in further understanding the safety and efficacy of potential AD prevention therapies targeting A?.
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Pleomorphic xanthoastrocytoma and oligodendroglioma: collision of 2 morphologically and genetically distinct anaplastic components.
J. Neurosurg.
PUBLISHED: 01-07-2011
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With the exception of oligoastrocytoma, mixed gliomas are rarely encountered, and the astrocytic component of mixed oligoastrocytoma is almost always fibrillary and diffusely infiltrative. Pleomorphic xanthoastrocytoma (PXA) has occasionally been described in conjunction with ganglioglioma, as well as in 1 case of oligodendroglioma. In this latter case, described by Perry et al., 1p/19q codeletions were not detected. The authors report on a 25-year-old woman with a combined PXA/oligodendroglioma in which concurrent 1p/19q codeletions were detected in the oligodendroglial component only. The patient presented with a 1-month history of headaches. Neuroimaging revealed a heterogeneous left temporal mass with focal enhancement, cystic changes, hemorrhage, and left-to-right midline shift. The patient underwent a craniotomy and gross-total resection. Pathological examination revealed a glial tumor composed of 2 apparently distinct components. The largest component exhibited a prominent fascicular, reticulin-rich, spindle cell arrangement admixed with areas of highly pleomorphic cells, with bizarre cytological features reminiscent of PXA. A smaller component was composed of cellular sheets and lobules of oligodendroglial cells. Both components were characterized by anaplastic features. Dual-color fluorescence in situ hybridization for 1p/19q codeletions was performed. Only the oligodendroglial component showed the combined 1p/19q deletions. This case represents the first instance in which PXA has been reported in conjunction with an oligodendroglioma exhibiting the "molecular signature" characteristic of oligodendroglial neoplasms. The different genetic alterations seen in the 2 components of this neoplasm argue in favor of a "collision tumor" rather than a mixed glioma of the same genotype.
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Long-term high-dose atorvastatin decreases brain oxidative and nitrosative stress in a preclinical model of Alzheimer disease: a novel mechanism of action.
Pharmacol. Res.
PUBLISHED: 11-10-2010
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Alzheimer disease (AD) is an age-related neurodegenerative disorder characterized by progressive memory loss, inability to perform the activities of daily living and personality changes. Unfortunately, drugs effective for this disease are limited to acetylcholinesterase inhibitors that do not impact disease pathogenesis. Statins, which belong to the class of cholesterol-reducing drugs, were proposed as novel agents useful in AD therapy, but the mechanism underlying their neuroprotective effect is still unknown. In this study, we show that atorvastatin may have antioxidant effects, in aged beagles, that represent a natural higher mammalian model of AD. Atorvastatin (80 mg/day for 14.5 months) significantly reduced lipoperoxidation, protein oxidation and nitration, and increased GSH levels in parietal cortex of aged beagles. This effect was specific for brain because it was not paralleled by a concomitant reduction in all these parameters in serum. In addition, atorvastatin slightly reduced the formation of cholesterol oxidation products in cortex but increased the 7-ketocholesterol/total cholesterol ratio in serum. We also found that increased oxidative damage in the parietal cortex was associated with poorer learning (visual discrimination task). Thus, a novel pharmacological effect of atorvastatin mediated by reducing oxidative damage may be one mechanism underlying benefits of this drug in AD.
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The role of fine-needle aspiration cytology in the surgical management of thyroid cancer.
ANZ J Surg
PUBLISHED: 10-26-2010
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Diagnosis of thyroid malignancy relies on clinical assessment, imaging and fine-needle aspiration cytology FNAC of thyroid nodules. The purpose of this study was to evaluate how effective synoptically reported FNAC is in clinical practice in diagnosing thyroid cancer. We also examined the effectiveness of using preoperative FNAC results to plan the type of operation for treating thyroid cancer.
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PE2GO: program evaluation of a physical activity program in elementary schools.
J Phys Act Health
PUBLISHED: 09-25-2010
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PE2GO is a self-contained physical education (PE) program that provides classroom teachers with the tools they need to lead developmentally appropriate PE lessons. The purpose of this study was to evaluate the PE2GO pilot programs in 6 school districts across the United States.
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Therapeutic targeting of the DNA mismatch repair pathway.
Clin. Cancer Res.
PUBLISHED: 09-07-2010
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The mismatch repair (MMR) pathway is involved in the removal of DNA base mismatches that arise either during DNA replication or are caused by DNA damage. Mutations in four genes involved in MMR, MSH2, MLH1, PMS2 and MSH6, predispose to a range of tumorigenic conditions, including hereditary nonpolyposis colon cancer, also known as Lynch syndrome. Here we discuss the canonical MMR pathway and the burgeoning evidence for noncanonical roles for the MMR genes, and highlight the therapeutic implications of MMR. In particular, we discuss how the DNA repair defect in MMR-deficient cancers could be exploited by the development of novel therapeutic strategies based on synthetic lethal approaches.
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A 41-year-old woman with von Hippel-Lindau and a cerebellar lesion.
Brain Pathol.
PUBLISHED: 05-05-2010
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A 41-year-old woman with a 12-year history of von Hippel-Lindau disease presented with progressive quadriparesis and difficulty swallowing. MRI revealed a well-circumscribed, partially cystic cerebellar neoplasm, consistent with hemangioblastoma. The tumor was resected and the diagnosis of hemangioblastoma confirmed. Embedded within the hemangioblastoma was a small focus of metastatic renal cell carcinoma (RCC). RCC metastatic to a CNS hemangioblastoma is the second most common type of tumor-to-tumor metastasis, which may be due to a number of factors. Proper immunostaining panels are required to clearly identify these cases since both tumor may have similar histology.
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A 61-year-old woman with osteomalacia and a thoracic spine lesion.
Brain Pathol.
PUBLISHED: 05-05-2010
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Phosphaturic mesenchymal tumor, mixed connective tissue variant (PMT-MCT) is a rare, largely benign, mesenchymal neoplasm almost invariably associated with oncogenic osteomalacia. It is generally found in the soft tissue and bone of the extremities. We report a case of a 61-year-old female with long-standing osteomalacia who was found to have PMT-MCT of the thoracic spine. There have been very few previously reported cases of PMT involving the spinal vertebrae and neuropathologists should be aware of this lesion. Recognition of PMT-MCT is critical for optimal patient care since complete surgical resection without additional therapy is curative.
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Assessing the capacity of state physical activity programs--a baseline perspective.
J Phys Act Health
PUBLISHED: 03-17-2010
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Physical inactivity is one of the top 3 risk factors associated with an increased prevalence of obesity and other chronic diseases. The public health infrastructure positions state health departments to address physical inactivity. To examine preparedness, all 50 health departments were assessed, using the 5 benchmarks developed by CDC for physical activity and public health practice, on their capacity to administer physical activity programs.
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Quantitative analysis of low-abundance peptides in HeLa cell cytoplasm by targeted liquid chromatography/mass spectrometry and stable isotope dilution: emphasising the distinction between peptide detection and peptide identification.
Rapid Commun. Mass Spectrom.
PUBLISHED: 03-11-2010
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We present the application of a targeted liquid chromatography/mass spectrometry (LC/MS) approach developed on a linear ion trap for the evaluation of the abundance of cytoplasmic proteins from a HeLa cell extract. Using a standard data-dependent approach, we identified some specific peptides from this extract which were also commercially available in their AQUA form (use for absolute quantitation). For some of the peptides, we observed a non-linear response between the intensity and the added quantity which was then fitted using a quadratic fit. All AQUA peptides spiked into a mix of 3 microg of the HeLa cell digest extract were detected down to 16 fmol. We placed an emphasis on peptide detection which, in this study, is performed using a combination of properties such as three specific Q3-like ion signatures (for a given Q1-like selection) and co-elution with the AQUA peptide counterparts. Detecting a peptide without necessarily identifying it using a search engine imposes less constraint in terms of tandem mass (MS/MS) spectra purity. An example is shown where a peptide is detected using those criteria but could not be identified by Mascot due to its lower abundance. To complement this observation, we used a cross-correlation analysis approach in order to separate two populations of MS/MS fragments based on differences in their elution patterns. Such an approach opens the door to new strategies to analyse lower intensity peptide fragments. An in silico analysis of the human trypsinosome allows the evaluation of how unique are the sets of features that we are using for peptide detection.
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Perioperative anxiety in children.
Paediatr Anaesth
PUBLISHED: 02-23-2010
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OBJECTIVES & AIM: The purpose of this investigation was to examine childrens anxiety across the perioperative setting.
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Mismatch repair deficient colorectal cancer in the era of personalized treatment.
Nat Rev Clin Oncol
PUBLISHED: 02-23-2010
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The molecular and genetic subtyping of cancer has allowed the emergence of individualized therapies. This approach could potentially deliver treatments that have both increased efficacy as well as reduced toxicity. A well-defined subtype of colorectal cancer (CRC) is characterized by a deficiency in the mismatch repair (MMR) pathway. MMR deficiency not only contributes to the pathogenesis of a large proportion of CRC, but also determines the response to many of the drugs that are frequently used to treat this disease. In this Review we describe the MMR deficient phenotype and discuss how a deficiency in this DNA repair process may impact on the management of CRC, including surgery, adjuvant chemotherapy and the choice of systemic agents for the palliation of advanced disease. We also discuss how the DNA repair defect in MMR deficient CRC could be exploited in the development of novel therapeutic strategies.
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Show me the money: financial incentives increase chlamydia screening rates among tertiary students: a pilot study.
Sex Health
PUBLISHED: 02-16-2010
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We hypothesise that text-messaging and financial incentives would increase tertiary student participation in chlamydia screening.
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What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.